IRESite_Id:486 characteristics of XIAP IRES studied in pbetaGAL/5'(-162)/CAT plasmid

IRESite: The database of experimentally verified IRES structures

The nucleic acid data:

IRESite Id: `486`	Version: `2`
Originaly submitted by: Václav Vopálenský
Reviewed by: Václav Vopálenský	Last change: `2008-07-16 23:25:42`

IRESite record type:

plasmid_with_promoter_and_putative_IRES_translationally_characterized

The shape of the nucleic acid molecule translated:
linear The quality of the mRNA/+RNA sequence:
3UTR_possibly_incomplete

The mRNA/+RNA description:

Putative in vivo CMV promoter-derived transcript produced from bicistronic plasmid pbetaGAL/5'(-162)/CAT which
comprises beta galactosidase and chloramphenicol acetyl transferase as the first and the second cistron,
respectively and the part of human XIAP mRNA (from nt -162 to nt -1 of the original sequence) mRNA.
The sequence ends at its 3'-end right after the poly(A) signal from BGH mRNA and thus the 3'-UTR might be
slightly wrong.

The mRNA/+RNA sequence represented in the +DNA notation:

agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc gagctcggat ccactaGTAA CGGCCGCCAG 
^1         ^11        ^21        ^31        ^41        ^51        ^61        ^71        ^81        ^91        

TGTGCTGGAA TTCTGCAGAT ATCCATCACA CTGGCGGCCG CAATTCCCGG GGATCGAAAG AGCCTGCTAA AGCAAAAAAG AAGTCACCat gtcgtttact 
^101       ^111       ^121       ^131       ^141       ^151       ^161       ^171       ^181       ^191       

ttgaccaaca agaacgtgat tttcgttgcc ggtctgggag gcattggtct ggacaccagc aaggagctgc tcaagcgcga tcccgtcgtt ttacaacgtc 
^201       ^211       ^221       ^231       ^241       ^251       ^261       ^271       ^281       ^291       

gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc 
^301       ^311       ^321       ^331       ^341       ^351       ^361       ^371       ^381       ^391       

ttcccaacag ttgcgcagcc tgaatggcga atggcgcttt gcctggtttc cggcaccaga agcggtgccg gaaagctggc tggagtgcga tcttcctgag 
^401       ^411       ^421       ^431       ^441       ^451       ^461       ^471       ^481       ^491       

gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg gttacgatgc gcccatctac accaacgtaa cctatcccat tacggtcaat ccgccgtttg 
^501       ^511       ^521       ^531       ^541       ^551       ^561       ^571       ^581       ^591       

ttcccacgga gaatccgacg ggttgttact cgctcacatt taatgttgat gaaagctggc tacaggaagg ccagacgcga attatttttg atggcgttaa 
^601       ^611       ^621       ^631       ^641       ^651       ^661       ^671       ^681       ^691       

ctcggcgttt catctgtggt gcaacgggcg ctgggtcggt tacggccagg acagtcgttt gccgtctgaa tttgacctga gcgcattttt acgcgccgga 
^701       ^711       ^721       ^731       ^741       ^751       ^761       ^771       ^781       ^791       

gaaaaccgcc tcgcggtgat ggtgctgcgt tggagtgacg gcagttatct ggaagatcag gatatgtggc ggatgagcgg cattttccgt gacgtctcgt 
^801       ^811       ^821       ^831       ^841       ^851       ^861       ^871       ^881       ^891       

tgctgcataa accgactaca caaatcagcg atttccatgt tgccactcgc tttaatgatg atttcagccg cgctgtactg gaggctgaag ttcagatgtg 
^901       ^911       ^921       ^931       ^941       ^951       ^961       ^971       ^981       ^991       

cggcgagttg cgtgactacc tacgggtaac agtttcttta tggcagggtg aaacgcaggt cgccagcggc accgcgcctt tcggcggtga aattatcgat 
^1001      ^1011      ^1021      ^1031      ^1041      ^1051      ^1061      ^1071      ^1081      ^1091      

gagcgtggtg gttatgccga tcgcgtcaca ctacgtctga acgtcgaaaa cccgaaactg tggagcgccg aaatcccgaa tctctatcgt gcggtggttg 
^1101      ^1111      ^1121      ^1131      ^1141      ^1151      ^1161      ^1171      ^1181      ^1191      

aactgcacac cgccgacggc acgctgattg aagcagaagc ctgcgatgtc ggtttccgcg aggtgcggat tgaaaatggt ctgctgctgc tgaacggcaa 
^1201      ^1211      ^1221      ^1231      ^1241      ^1251      ^1261      ^1271      ^1281      ^1291      

gccgttgctg attcgaggcg ttaaccgtca cgagcatcat cctctgcatg gtcaggtcat ggatgagcag acgatggtgc aggatatcct gctgatgaag 
^1301      ^1311      ^1321      ^1331      ^1341      ^1351      ^1361      ^1371      ^1381      ^1391      

cagaacaact ttaacgccgt gcgctgttcg cattatccga accatccgct gtggtacacg ctgtgcgacc gctacggcct gtatgtggtg gatgaagcca 
^1401      ^1411      ^1421      ^1431      ^1441      ^1451      ^1461      ^1471      ^1481      ^1491      

atattgaaac ccacggcatg gtgccaatga atcgtctgac cgatgatccg cgctggctac cggcgatgag cgaacgcgta acgcgaatgg tgcagcgcga 
^1501      ^1511      ^1521      ^1531      ^1541      ^1551      ^1561      ^1571      ^1581      ^1591      

tcgtaatcac ccgagtgtga tcatctggtc gctggggaat gaatcaggcc acggcgctaa tcacgacgcg ctgtatcgct ggatcaaatc tgtcgatcct 
^1601      ^1611      ^1621      ^1631      ^1641      ^1651      ^1661      ^1671      ^1681      ^1691      

tcccgcccgg tgcagtatga aggcggcgga gccgacacca cggccaccga tattatttgc ccgatgtacg cgcgcgtgga tgaagaccag cccttcccgg 
^1701      ^1711      ^1721      ^1731      ^1741      ^1751      ^1761      ^1771      ^1781      ^1791      

ctgtgccgaa atggtccatc aaaaaatggc tttcgctacc tggagagacg cgcccgctga tcctttgcga atacgcccac gcgatgggta acagtcttgg 
^1801      ^1811      ^1821      ^1831      ^1841      ^1851      ^1861      ^1871      ^1881      ^1891      

cggtttcgct aaatactggc aggcgtttcg tcagtatccc cgtttacagg gcggcttcgt ctgggactgg gtggatcagt cgctgattaa atatgatgaa 
^1901      ^1911      ^1921      ^1931      ^1941      ^1951      ^1961      ^1971      ^1981      ^1991      

aacggcaacc cgtggtcggc ttacggcggt gattttggcg atacgccgaa cgatcgccag ttctgtatga acggtctggt ctttgccgac cgcacgccgc 
^2001      ^2011      ^2021      ^2031      ^2041      ^2051      ^2061      ^2071      ^2081      ^2091      

atccagcgct gacggaagca aaacaccagc agcagttttt ccagttccgt ttatccgggc aaaccatcga agtgaccagc gaatacctgt tccgtcatag 
^2101      ^2111      ^2121      ^2131      ^2141      ^2151      ^2161      ^2171      ^2181      ^2191      

cgataacgag ctcctgcact ggatggtggc gctggatggt aagccgctgg caagcggtga agtgcctctg gatgtcgctc cacaaggtaa acagttgatt 
^2201      ^2211      ^2221      ^2231      ^2241      ^2251      ^2261      ^2271      ^2281      ^2291      

gaactgcctg aactaccgca gccggagagc gccgggcaac tctggctcac agtacgcgta gtgcaaccga acgcgaccgc atggtcagaa gccgggcaca 
^2301      ^2311      ^2321      ^2331      ^2341      ^2351      ^2361      ^2371      ^2381      ^2391      

tcagcgcctg gcagcagtgg cgtctggcgg aaaacctcag tgtgacgctc cccgccgcgt cccacgccat cccgcatctg accaccagcg aaatggattt 
^2401      ^2411      ^2421      ^2431      ^2441      ^2451      ^2461      ^2471      ^2481      ^2491      

ttgcatcgag ctgggtaata agcgttggca atttaaccgc cagtcaggct ttctttcaca gatgtggatt ggcgataaaa aacaactgct gacgccgctg 
^2501      ^2511      ^2521      ^2531      ^2541      ^2551      ^2561      ^2571      ^2581      ^2591      

cgcgatcagt tcacccgtgc accgctggat aacgacattg gcgtaagtga agcgacccgc attgacccta acgcctgggt cgaacgctgg aaggcggcgg 
^2601      ^2611      ^2621      ^2631      ^2641      ^2651      ^2661      ^2671      ^2681      ^2691      

gccattacca ggccgaagca gcgttgttgc agtgcacggc agatacactt gctgatgcgg tgctgattac gaccgctcac gcgtggcagc atcaggggaa 
^2701      ^2711      ^2721      ^2731      ^2741      ^2751      ^2761      ^2771      ^2781      ^2791      

aaccttattt atcagccgga aaacctaccg gattgatggt agtggtcaaa tggcgattac cgttgatgtt gaagtggcga gcgatacacc gcatccggcg 
^2801      ^2811      ^2821      ^2831      ^2841      ^2851      ^2861      ^2871      ^2881      ^2891      

cggattggcc tgaactgcca gctggcgcag gtagcagagc gggtaaactg gctcggatta gggccgcaag aaaactatcc cgaccgcctt actgccgcct 
^2901      ^2911      ^2921      ^2931      ^2941      ^2951      ^2961      ^2971      ^2981      ^2991      

gttttgaccg ctgggatctg ccattgtcag acatgtatac cccgtacgtc ttcccgagcg aaaacggtct gcgctgcggg acgcgcgaat tgaattatgg 
^3001      ^3011      ^3021      ^3031      ^3041      ^3051      ^3061      ^3071      ^3081      ^3091      

cccacaccag tggcgcggcg acttccagtt caacatcagc cgctacagtc aacagcaact gatggaaacc agccatcgcc atctgctgca cgcggaagaa 
^3101      ^3111      ^3121      ^3131      ^3141      ^3151      ^3161      ^3171      ^3181      ^3191      

ggcacatggc tgaatatcga cggtttccat atggggattg gtggcgacga ctcctggagc ccgtcagtat cggcggaatt acagctgagc gccggtcgct 
^3201      ^3211      ^3221      ^3231      ^3241      ^3251      ^3261      ^3271      ^3281      ^3291      

accattacca gttggtctgg tgtcaaaaat aaTAATAACC GGGCAGGCCA TGTCTGCCCG TATTTCGCGT AAGGAAATCC ATTATGTACT ATTTAAAAAA 
^3301      ^3311      ^3321      ^3331      ^3341      ^3351      ^3361      ^3371      ^3381      ^3391      

CACAAACTTT TGGATGTTCG GTTTATTCTT TTTCTTTTAC TTTTTTATCA TGGGAGCCTA CTTCCCGTTT TTCCCGATTT GGCTACATGA CATCAACCAT 
^3401      ^3411      ^3421      ^3431      ^3441      ^3451      ^3461      ^3471      ^3481      ^3491      

ATCAGCAAAA GTGATACGGG TATTATTTTT GCCGCTATTT CTCTGTTCTC GCTATTATTC CAACCGCTGT TTGGTCTGCT TTCTGACAAA CTCGGCCTCG 
^3501      ^3511      ^3521      ^3531      ^3541      ^3551      ^3561      ^3571      ^3581      ^3591      

ACTCTAGGCG GCCGCTCGAG ATTAGAATGT TTCTTAGCGG TCGTGTAGTT ATTTTTATGT CATAAGTGGA TAATTTGTTA GCTCCTATAA CAAAAGTCTG 
^3601      ^3611      ^3621      ^3631      ^3641      ^3651      ^3661      ^3671      ^3681      ^3691      

TTGCTTGTGT TTCACATTTT GGATTTCCTA ATATAATGTT CTCTTTTTAG AAAAGGTGGA CAAGTCCTAT TTTCAAGAGA AGCTCGAGCT AGAGGATCTG 
^3701      ^3711      ^3721      ^3731      ^3741      ^3751      ^3761      ^3771      ^3781      ^3791      

AGCTTGGCGA GATTTTCAGG AGCTAAGGAA GCTAAAatgg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt 
^3801      ^3811      ^3821      ^3831      ^3841      ^3851      ^3861      ^3871      ^3881      ^3891      

ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt 
^3901      ^3911      ^3921      ^3931      ^3941      ^3951      ^3961      ^3971      ^3981      ^3991      

ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg aattccgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt 
^4001      ^4011      ^4021      ^4031      ^4041      ^4051      ^4061      ^4071      ^4081      ^4091      

cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc 
^4101      ^4111      ^4121      ^4131      ^4141      ^4151      ^4161      ^4171      ^4181      ^4191      

aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag 
^4201      ^4211      ^4221      ^4231      ^4241      ^4251      ^4261      ^4271      ^4281      ^4291      

ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg 
^4301      ^4311      ^4321      ^4331      ^4341      ^4351      ^4361      ^4371      ^4381      ^4391      

attcaggttc atcatgccgt ctgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttt 
^4401      ^4411      ^4421      ^4431      ^4441      ^4451      ^4461      ^4471      ^4481      ^4491      

tttaaggcag ttattggtgc ccttaaacgc ctggtgctac gcctgaataa gtgataataa gcggatgaat ggcagaaatt cgccggatct ttgtgaagga 
^4501      ^4511      ^4521      ^4531      ^4541      ^4551      ^4561      ^4571      ^4581      ^4591      

accttacttc tgtggtgtga cataattgga caaactacct acagagattt aaagctctaa ggtaaatata aaatttttaa gtgtataatg tgttaaacta 
^4601      ^4611      ^4621      ^4631      ^4641      ^4651      ^4661      ^4671      ^4681      ^4691      

ctgattctaa ttgtttgtgt attttagatt ccaacctatg gaactgatga atgggagcag tggtggaatg cctttaatga ggaaaacctg ttttgctcag 
^4701      ^4711      ^4721      ^4731      ^4741      ^4751      ^4761      ^4771      ^4781      ^4791      

aagaaatgcc atctagtgat gatgaggcta ctgctgactc tcaacattct actcctccaa aaaagaagag aaaggtagaa gaccccaagg actttccttc 
^4801      ^4811      ^4821      ^4831      ^4841      ^4851      ^4861      ^4871      ^4881      ^4891      

agaattgcta agttttttga gtcatgctgt gtttagtaat agaactcttg cttgctttgc tatttacacc acaaaggaaa aagctgcact gctatacaag 
^4901      ^4911      ^4921      ^4931      ^4941      ^4951      ^4961      ^4971      ^4981      ^4991      

aaaattatgg aaaaatattc tgtaaccttt ataagtaggc ataacagtta taatcataac atactgtttt ttcttactcc acacaggcat agagtgtctg 
^5001      ^5011      ^5021      ^5031      ^5041      ^5051      ^5061      ^5071      ^5081      ^5091      

ctattaataa ctatgctcaa aaattgtgta cctttagctt tttaatttgt aaaggggtta ataaggaata tttgatgtat agtgccttga ctagagatca 
^5101      ^5111      ^5121      ^5131      ^5141      ^5151      ^5161      ^5171      ^5181      ^5191      

taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt 
^5201      ^5211      ^5221      ^5231      ^5241      ^5251      ^5261      ^5271      ^5281      ^5291      

taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 
^5301      ^5311      ^5321      ^5331      ^5341      ^5351      ^5361      ^5371      ^5381      ^5391      

tccaaactca tcaatgtatc ttatcatgtc tgcctagagg gccctattct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt 
^5401      ^5411      ^5421      ^5431      ^5441      ^5451      ^5461      ^5471      ^5481      ^5491      

ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataa
^5501      ^5511      ^5521      ^5531      ^5541      ^5551      ^5561      ^5571      ^5581

Credibility of mRNA sequence:

reverse_engineered_sequence_and_should_match_experiment_maybe_except_3UTR

The name of the plasmid:
pbetaGAL/5'(-162)/CAT The name of the promoter used to express this mRNA:
CMV

Description of the plasmid (facultative for promoter-less plasmid records):
Part of the 5' UTR from human XIAP mRNA (from nt -162 to nt -1 of the original sequence) has been cloned between the beta galactosidase and chloramphenicol acetyl transferase reporter genes.

The in vivo produced transcripts are heterogeneous (due to any of promoter?/splicing?/cleavage?/breakage?):

no

The in vivo produced heterogeneous transcripts occur due to alternative splicing:

no

A promoter reported in cDNA corresponding to IRES sequence:

no

Summary of possible issues when IRES cDNA is experimentally transcribed in vivo:

Summary of experiments studying integrity of the in vivo transcripts in a particular host:

Integrity (uniformity) of mRNA tested using Northern-blot:
not_tested

Integrity (uniformity) of mRNA tested using RNase protection:
not_tested

Integrity (uniformity) of mRNA tested using 5'-RACE:
not_tested

Integrity (uniformity) of mRNA tested using primer extension :
not_tested

Integrity (uniformity) of mRNA tested using RT-PCR:
not_tested

Integrity (uniformity) of mRNA tested using real-time quantitative polymerase chain reaction (rtqPCR):
homogeneous_population_of_molecules_confirmed

Integrity (uniformity) of mRNA tested using RNAi:
not_tested

Integrity (uniformity) of mRNA tested using S1 nuclease mapping:
not_tested

Cryptic promoter presence was confirmed by expression from a promoter-less plasmid:
no_promoter_confirmed

Cryptic promoter presence was confirmed in an experimental setup involving inducible promoter:
not_tested

Integrity (uniformity) of mRNA molecules or possible promoter presence expressed in vivo was tested using another method, please specify in Remarks:
not_tested

The organism used:

Homo sapiens HEK293 (ATCC CRL-1573)

The abbreviated name of the donor gene or virus from which this IRES was excised and inserted into the plasmid:
XIAP

The origin of IRES in the plasmid:

nuclear

The donor organism of the IRES segment:

Homo sapiens

The DNA sequence of the plasmid in (+) orientation annotated by its secondary structure:

TTTGAATGTA TTTAGAAAAA TAAACAAATA GGGGTTCCGC GCACATTTCC CCGAAAAGTG CCACCTGACG TCGACGGATC GGGAGATCTC CCGATCCCCT 
^1         ^11        ^21        ^31        ^41        ^51        ^61        ^71        ^81        ^91        

ATGGTCGACT CTCAGTACAA TCTGCTCTGA TGCCGCATAG TTAAGCCAGT ATCTGCTCCC TGCTTGTGTG TTGGAGGTCG CTGAGTAGTG CGCGAGCAAA 
^101       ^111       ^121       ^131       ^141       ^151       ^161       ^171       ^181       ^191       

atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata 
^201       ^211       ^221       ^231       ^241       ^251       ^261       ^271       ^281       ^291       

cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt 
^301       ^311       ^321       ^331       ^341       ^351       ^361       ^371       ^381       ^391       

aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt 
^401       ^411       ^421       ^431       ^441       ^451       ^461       ^471       ^481       ^491       

caatgggtgg actatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc 
^501       ^511       ^521       ^531       ^541       ^551       ^561       ^571       ^581       ^591       

ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg 
^601       ^611       ^621       ^631       ^641       ^651       ^661       ^671       ^681       ^691       

gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa 
^701       ^711       ^721       ^731       ^741       ^751       ^761       ^771       ^781       ^791       

cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa 
^801       ^811       ^821       ^831       ^841       ^851       ^861       ^871       ^881       ^891       

ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagcttggta ccgagctcgg atccactaGT AACGGCCGCC 
^901       ^911       ^921       ^931       ^941       ^951       ^961       ^971       ^981       ^991       

AGTGTGCTGG AATTCTGCAG ATATCCATCA CACTGGCGGC CGCAATTCCC GGGGATCGAA AGAGCCTGCT AAAGCAAAAA AGAAGTCACC atgtcgttta 
^1001      ^1011      ^1021      ^1031      ^1041      ^1051      ^1061      ^1071      ^1081      ^1091      

ctttgaccaa caagaacgtg attttcgttg ccggtctggg aggcattggt ctggacacca gcaaggagct gctcaagcgc gatcccgtcg ttttacaacg 
^1101      ^1111      ^1121      ^1131      ^1141      ^1151      ^1161      ^1171      ^1181      ^1191      

tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc 
^1201      ^1211      ^1221      ^1231      ^1241      ^1251      ^1261      ^1271      ^1281      ^1291      

ccttcccaac agttgcgcag cctgaatggc gaatggcgct ttgcctggtt tccggcacca gaagcggtgc cggaaagctg gctggagtgc gatcttcctg 
^1301      ^1311      ^1321      ^1331      ^1341      ^1351      ^1361      ^1371      ^1381      ^1391      

aggccgatac tgtcgtcgtc ccctcaaact ggcagatgca cggttacgat gcgcccatct acaccaacgt aacctatccc attacggtca atccgccgtt 
^1401      ^1411      ^1421      ^1431      ^1441      ^1451      ^1461      ^1471      ^1481      ^1491      

tgttcccacg gagaatccga cgggttgtta ctcgctcaca tttaatgttg atgaaagctg gctacaggaa ggccagacgc gaattatttt tgatggcgtt 
^1501      ^1511      ^1521      ^1531      ^1541      ^1551      ^1561      ^1571      ^1581      ^1591      

aactcggcgt ttcatctgtg gtgcaacggg cgctgggtcg gttacggcca ggacagtcgt ttgccgtctg aatttgacct gagcgcattt ttacgcgccg 
^1601      ^1611      ^1621      ^1631      ^1641      ^1651      ^1661      ^1671      ^1681      ^1691      

gagaaaaccg cctcgcggtg atggtgctgc gttggagtga cggcagttat ctggaagatc aggatatgtg gcggatgagc ggcattttcc gtgacgtctc 
^1701      ^1711      ^1721      ^1731      ^1741      ^1751      ^1761      ^1771      ^1781      ^1791      

gttgctgcat aaaccgacta cacaaatcag cgatttccat gttgccactc gctttaatga tgatttcagc cgcgctgtac tggaggctga agttcagatg 
^1801      ^1811      ^1821      ^1831      ^1841      ^1851      ^1861      ^1871      ^1881      ^1891      

tgcggcgagt tgcgtgacta cctacgggta acagtttctt tatggcaggg tgaaacgcag gtcgccagcg gcaccgcgcc tttcggcggt gaaattatcg 
^1901      ^1911      ^1921      ^1931      ^1941      ^1951      ^1961      ^1971      ^1981      ^1991      

atgagcgtgg tggttatgcc gatcgcgtca cactacgtct gaacgtcgaa aacccgaaac tgtggagcgc cgaaatcccg aatctctatc gtgcggtggt 
^2001      ^2011      ^2021      ^2031      ^2041      ^2051      ^2061      ^2071      ^2081      ^2091      

tgaactgcac accgccgacg gcacgctgat tgaagcagaa gcctgcgatg tcggtttccg cgaggtgcgg attgaaaatg gtctgctgct gctgaacggc 
^2101      ^2111      ^2121      ^2131      ^2141      ^2151      ^2161      ^2171      ^2181      ^2191      

aagccgttgc tgattcgagg cgttaaccgt cacgagcatc atcctctgca tggtcaggtc atggatgagc agacgatggt gcaggatatc ctgctgatga 
^2201      ^2211      ^2221      ^2231      ^2241      ^2251      ^2261      ^2271      ^2281      ^2291      

agcagaacaa ctttaacgcc gtgcgctgtt cgcattatcc gaaccatccg ctgtggtaca cgctgtgcga ccgctacggc ctgtatgtgg tggatgaagc 
^2301      ^2311      ^2321      ^2331      ^2341      ^2351      ^2361      ^2371      ^2381      ^2391      

caatattgaa acccacggca tggtgccaat gaatcgtctg accgatgatc cgcgctggct accggcgatg agcgaacgcg taacgcgaat ggtgcagcgc 
^2401      ^2411      ^2421      ^2431      ^2441      ^2451      ^2461      ^2471      ^2481      ^2491      

gatcgtaatc acccgagtgt gatcatctgg tcgctgggga atgaatcagg ccacggcgct aatcacgacg cgctgtatcg ctggatcaaa tctgtcgatc 
^2501      ^2511      ^2521      ^2531      ^2541      ^2551      ^2561      ^2571      ^2581      ^2591      

cttcccgccc ggtgcagtat gaaggcggcg gagccgacac cacggccacc gatattattt gcccgatgta cgcgcgcgtg gatgaagacc agcccttccc 
^2601      ^2611      ^2621      ^2631      ^2641      ^2651      ^2661      ^2671      ^2681      ^2691      

ggctgtgccg aaatggtcca tcaaaaaatg gctttcgcta cctggagaga cgcgcccgct gatcctttgc gaatacgccc acgcgatggg taacagtctt 
^2701      ^2711      ^2721      ^2731      ^2741      ^2751      ^2761      ^2771      ^2781      ^2791      

ggcggtttcg ctaaatactg gcaggcgttt cgtcagtatc cccgtttaca gggcggcttc gtctgggact gggtggatca gtcgctgatt aaatatgatg 
^2801      ^2811      ^2821      ^2831      ^2841      ^2851      ^2861      ^2871      ^2881      ^2891      

aaaacggcaa cccgtggtcg gcttacggcg gtgattttgg cgatacgccg aacgatcgcc agttctgtat gaacggtctg gtctttgccg accgcacgcc 
^2901      ^2911      ^2921      ^2931      ^2941      ^2951      ^2961      ^2971      ^2981      ^2991      

gcatccagcg ctgacggaag caaaacacca gcagcagttt ttccagttcc gtttatccgg gcaaaccatc gaagtgacca gcgaatacct gttccgtcat 
^3001      ^3011      ^3021      ^3031      ^3041      ^3051      ^3061      ^3071      ^3081      ^3091      

agcgataacg agctcctgca ctggatggtg gcgctggatg gtaagccgct ggcaagcggt gaagtgcctc tggatgtcgc tccacaaggt aaacagttga 
^3101      ^3111      ^3121      ^3131      ^3141      ^3151      ^3161      ^3171      ^3181      ^3191      

ttgaactgcc tgaactaccg cagccggaga gcgccgggca actctggctc acagtacgcg tagtgcaacc gaacgcgacc gcatggtcag aagccgggca 
^3201      ^3211      ^3221      ^3231      ^3241      ^3251      ^3261      ^3271      ^3281      ^3291      

catcagcgcc tggcagcagt ggcgtctggc ggaaaacctc agtgtgacgc tccccgccgc gtcccacgcc atcccgcatc tgaccaccag cgaaatggat 
^3301      ^3311      ^3321      ^3331      ^3341      ^3351      ^3361      ^3371      ^3381      ^3391      

ttttgcatcg agctgggtaa taagcgttgg caatttaacc gccagtcagg ctttctttca cagatgtgga ttggcgataa aaaacaactg ctgacgccgc 
^3401      ^3411      ^3421      ^3431      ^3441      ^3451      ^3461      ^3471      ^3481      ^3491      

tgcgcgatca gttcacccgt gcaccgctgg ataacgacat tggcgtaagt gaagcgaccc gcattgaccc taacgcctgg gtcgaacgct ggaaggcggc 
^3501      ^3511      ^3521      ^3531      ^3541      ^3551      ^3561      ^3571      ^3581      ^3591      

gggccattac caggccgaag cagcgttgtt gcagtgcacg gcagatacac ttgctgatgc ggtgctgatt acgaccgctc acgcgtggca gcatcagggg 
^3601      ^3611      ^3621      ^3631      ^3641      ^3651      ^3661      ^3671      ^3681      ^3691      

aaaaccttat ttatcagccg gaaaacctac cggattgatg gtagtggtca aatggcgatt accgttgatg ttgaagtggc gagcgataca ccgcatccgg 
^3701      ^3711      ^3721      ^3731      ^3741      ^3751      ^3761      ^3771      ^3781      ^3791      

cgcggattgg cctgaactgc cagctggcgc aggtagcaga gcgggtaaac tggctcggat tagggccgca agaaaactat cccgaccgcc ttactgccgc 
^3801      ^3811      ^3821      ^3831      ^3841      ^3851      ^3861      ^3871      ^3881      ^3891      

ctgttttgac cgctgggatc tgccattgtc agacatgtat accccgtacg tcttcccgag cgaaaacggt ctgcgctgcg ggacgcgcga attgaattat 
^3901      ^3911      ^3921      ^3931      ^3941      ^3951      ^3961      ^3971      ^3981      ^3991      

ggcccacacc agtggcgcgg cgacttccag ttcaacatca gccgctacag tcaacagcaa ctgatggaaa ccagccatcg ccatctgctg cacgcggaag 
^4001      ^4011      ^4021      ^4031      ^4041      ^4051      ^4061      ^4071      ^4081      ^4091      

aaggcacatg gctgaatatc gacggtttcc atatggggat tggtggcgac gactcctgga gcccgtcagt atcggcggaa ttacagctga gcgccggtcg 
^4101      ^4111      ^4121      ^4131      ^4141      ^4151      ^4161      ^4171      ^4181      ^4191      

ctaccattac cagttggtct ggtgtcaaaa ataaTAATAA CCGGGCAGGC CATGTCTGCC CGTATTTCGC GTAAGGAAAT CCATTATGTA CTATTTAAAA 
^4201      ^4211      ^4221      ^4231      ^4241      ^4251      ^4261      ^4271      ^4281      ^4291      

AACACAAACT TTTGGATGTT CGGTTTATTC TTTTTCTTTT ACTTTTTTAT CATGGGAGCC TACTTCCCGT TTTTCCCGAT TTGGCTACAT GACATCAACC 
^4301      ^4311      ^4321      ^4331      ^4341      ^4351      ^4361      ^4371      ^4381      ^4391      

ATATCAGCAA AAGTGATACG GGTATTATTT TTGCCGCTAT TTCTCTGTTC TCGCTATTAT TCCAACCGCT GTTTGGTCTG CTTTCTGACA AACTCGGCCT 
^4401      ^4411      ^4421      ^4431      ^4441      ^4451      ^4461      ^4471      ^4481      ^4491      

CGACTCTAGG CGGCCGCTCG AGATTAGAAT GTTTCTTAGC GGTCGTGTAG TTATTTTTAT GTCATAAGTG GATAATTTGT TAGCTCCTAT AACAAAAGTC 
^4501      ^4511      ^4521      ^4531      ^4541      ^4551      ^4561      ^4571      ^4581      ^4591      

TGTTGCTTGT GTTTCACATT TTGGATTTCC TAATATAATG TTCTCTTTTT AGAAAAGGTG GACAAGTCCT ATTTTCAAGA GAAGCTCGAG CTAGAGGATC 
^4601      ^4611      ^4621      ^4631      ^4641      ^4651      ^4661      ^4671      ^4681      ^4691      

TGAGCTTGGC GAGATTTTCA GGAGCTAAGG AAGCTAAAat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca 
^4701      ^4711      ^4721      ^4731      ^4741      ^4751      ^4761      ^4771      ^4781      ^4791      

ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag 
^4801      ^4811      ^4821      ^4831      ^4841      ^4851      ^4861      ^4871      ^4881      ^4891      

ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattccgt atggcaatga aagacggtga gctggtgata tgggatagtg 
^4901      ^4911      ^4921      ^4931      ^4941      ^4951      ^4961      ^4971      ^4981      ^4991      

ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc 
^5001      ^5011      ^5021      ^5031      ^5041      ^5051      ^5061      ^5071      ^5081      ^5091      

gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc 
^5101      ^5111      ^5121      ^5131      ^5141      ^5151      ^5161      ^5171      ^5181      ^5191      

agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg 
^5201      ^5211      ^5221      ^5231      ^5241      ^5251      ^5261      ^5271      ^5281      ^5291      

cgattcaggt tcatcatgcc gtctgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaatt 
^5301      ^5311      ^5321      ^5331      ^5341      ^5351      ^5361      ^5371      ^5381      ^5391      

tttttaaggc agttattggt gcccttaaac gcctggtgct acgcctgaat aagtgataat aagcggatga atggcagaaa ttcgccggat ctttgtgaag 
^5401      ^5411      ^5421      ^5431      ^5441      ^5451      ^5461      ^5471      ^5481      ^5491      

gaaccttact tctgtggtgt gacataattg gacaaactac ctacagagat ttaaagctct aaggtaaata taaaattttt aagtgtataa tgtgttaaac 
^5501      ^5511      ^5521      ^5531      ^5541      ^5551      ^5561      ^5571      ^5581      ^5591      

tactgattct aattgtttgt gtattttaga ttccaaccta tggaactgat gaatgggagc agtggtggaa tgcctttaat gaggaaaacc tgttttgctc 
^5601      ^5611      ^5621      ^5631      ^5641      ^5651      ^5661      ^5671      ^5681      ^5691      

agaagaaatg ccatctagtg atgatgaggc tactgctgac tctcaacatt ctactcctcc aaaaaagaag agaaaggtag aagaccccaa ggactttcct 
^5701      ^5711      ^5721      ^5731      ^5741      ^5751      ^5761      ^5771      ^5781      ^5791      

tcagaattgc taagtttttt gagtcatgct gtgtttagta atagaactct tgcttgcttt gctatttaca ccacaaagga aaaagctgca ctgctataca 
^5801      ^5811      ^5821      ^5831      ^5841      ^5851      ^5861      ^5871      ^5881      ^5891      

agaaaattat ggaaaaatat tctgtaacct ttataagtag gcataacagt tataatcata acatactgtt ttttcttact ccacacaggc atagagtgtc 
^5901      ^5911      ^5921      ^5931      ^5941      ^5951      ^5961      ^5971      ^5981      ^5991      

tgctattaat aactatgctc aaaaattgtg tacctttagc tttttaattt gtaaaggggt taataaggaa tatttgatgt atagtgcctt gactagagat 
^6001      ^6011      ^6021      ^6031      ^6041      ^6051      ^6061      ^6071      ^6081      ^6091      

cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt 
^6101      ^6111      ^6121      ^6131      ^6141      ^6151      ^6161      ^6171      ^6181      ^6191      

gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt 
^6201      ^6211      ^6221      ^6231      ^6241      ^6251      ^6261      ^6271      ^6281      ^6291      

tgtccaaact catcaatgta tcttatcatg tctgcctaga gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc 
^6301      ^6311      ^6321      ^6331      ^6341      ^6351      ^6361      ^6371      ^6381      ^6391      

ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt 
^6401      ^6411      ^6421      ^6431      ^6441      ^6451      ^6461      ^6471      ^6481      ^6491      

gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg 
^6501      ^6511      ^6521      ^6531      ^6541      ^6551      ^6561      ^6571      ^6581      ^6591      

atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt 
^6601      ^6611      ^6621      ^6631      ^6641      ^6651      ^6661      ^6671      ^6681      ^6691      

ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 
^6701      ^6711      ^6721      ^6731      ^6741      ^6751      ^6761      ^6771      ^6781      ^6791      

cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg 
^6801      ^6811      ^6821      ^6831      ^6841      ^6851      ^6861      ^6871      ^6881      ^6891      

ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat 
^6901      ^6911      ^6921      ^6931      ^6941      ^6951      ^6961      ^6971      ^6981      ^6991      

ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt 
^7001      ^7011      ^7021      ^7031      ^7041      ^7051      ^7061      ^7071      ^7081      ^7091      

ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccaggcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa 
^7101      ^7111      ^7121      ^7131      ^7141      ^7151      ^7161      ^7171      ^7181      ^7191      

agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac 
^7201      ^7211      ^7221      ^7231      ^7241      ^7251      ^7261      ^7271      ^7281      ^7291      

tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag 
^7301      ^7311      ^7321      ^7331      ^7341      ^7351      ^7361      ^7371      ^7381      ^7391      

tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc 
^7401      ^7411      ^7421      ^7431      ^7441      ^7451      ^7461      ^7471      ^7481      ^7491      

gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga 
^7501      ^7511      ^7521      ^7531      ^7541      ^7551      ^7561      ^7571      ^7581      ^7591      

tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg 
^7601      ^7611      ^7621      ^7631      ^7641      ^7651      ^7661      ^7671      ^7681      ^7691      

ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc 
^7701      ^7711      ^7721      ^7731      ^7741      ^7751      ^7761      ^7771      ^7781      ^7791      

aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt 
^7801      ^7811      ^7821      ^7831      ^7841      ^7851      ^7861      ^7871      ^7881      ^7891      

cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg 
^7901      ^7911      ^7921      ^7931      ^7941      ^7951      ^7961      ^7971      ^7981      ^7991      

ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa 
^8001      ^8011      ^8021      ^8031      ^8041      ^8051      ^8061      ^8071      ^8081      ^8091      

atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg 
^8101      ^8111      ^8121      ^8131      ^8141      ^8151      ^8161      ^8171      ^8181      ^8191      

cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg 
^8201      ^8211      ^8221      ^8231      ^8241      ^8251      ^8261      ^8271      ^8281      ^8291      

ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat 
^8301      ^8311      ^8321      ^8331      ^8341      ^8351      ^8361      ^8371      ^8381      ^8391      

cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac 
^8401      ^8411      ^8421      ^8431      ^8441      ^8451      ^8461      ^8471      ^8481      ^8491      

aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 
^8501      ^8511      ^8521      ^8531      ^8541      ^8551      ^8561      ^8571      ^8581      ^8591      

gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 
^8601      ^8611      ^8621      ^8631      ^8641      ^8651      ^8661      ^8671      ^8681      ^8691      

cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 
^8701      ^8711      ^8721      ^8731      ^8741      ^8751      ^8761      ^8771      ^8781      ^8791      

tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 
^8801      ^8811      ^8821      ^8831      ^8841      ^8851      ^8861      ^8871      ^8881      ^8891      

ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 
^8901      ^8911      ^8921      ^8931      ^8941      ^8951      ^8961      ^8971      ^8981      ^8991      

gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 
^9001      ^9011      ^9021      ^9031      ^9041      ^9051      ^9061      ^9071      ^9081      ^9091      

tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 
^9101      ^9111      ^9121      ^9131      ^9141      ^9151      ^9161      ^9171      ^9181      ^9191      

ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 
^9201      ^9211      ^9221      ^9231      ^9241      ^9251      ^9261      ^9271      ^9281      ^9291      

cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 
^9301      ^9311      ^9321      ^9331      ^9341      ^9351      ^9361      ^9371      ^9381      ^9391      

ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct 
^9401      ^9411      ^9421      ^9431      ^9441      ^9451      ^9461      ^9471      ^9481      ^9491      

gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 
^9501      ^9511      ^9521      ^9531      ^9541      ^9551      ^9561      ^9571      ^9581      ^9591      

gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 
^9601      ^9611      ^9621      ^9631      ^9641      ^9651      ^9661      ^9671      ^9681      ^9691      

gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 
^9701      ^9711      ^9721      ^9731      ^9741      ^9751      ^9761      ^9771      ^9781      ^9791      

gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 
^9801      ^9811      ^9821      ^9831      ^9841      ^9851      ^9861      ^9871      ^9881      ^9891      

ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 
^9901      ^9911      ^9921      ^9931      ^9941      ^9951      ^9961      ^9971      ^9981      ^9991      

gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 
^10001     ^10011     ^10021     ^10031     ^10041     ^10051     ^10061     ^10071     ^10081     ^10091     

acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 
^10101     ^10111     ^10121     ^10131     ^10141     ^10151     ^10161     ^10171     ^10181     ^10191     

ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 
^10201     ^10211     ^10221     ^10231     ^10241     ^10251     ^10261     ^10271     ^10281     ^10291     

cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 
^10301     ^10311     ^10321     ^10331     ^10341     ^10351     ^10361     ^10371     ^10381     ^10391     

gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 
^10401     ^10411     ^10421     ^10431     ^10441     ^10451     ^10461     ^10471     ^10481     ^10491     

gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 
^10501     ^10511     ^10521     ^10531     ^10541     ^10551     ^10561     ^10571     ^10581     ^10591     

acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg 
^10601     ^10611     ^10621     ^10631     ^10641     ^10651     ^10661     ^10671     ^10681     ^10691     

gttattgtct catgagcgga tacata
^10701     ^10711     ^10721

GenBank formatted file with annotated plasmid sequence hyperlinked from vector image map:

The total number of notable open-reading frames (ORFs):

2

Notable Open-Reading Frames (ORFs; protein coding regions) in the mRNA/+RNA sequence:

ORF

ORF position:

1

Version: `0`
Originaly submitted by: Václav Vopálenský	Reviewed by: Václav Vopálenský

The abbreviated name of this ORF/gene:
LacZ

The description of the protein encoded in this ORF:
beta galactosidase

The translational frameshift (ribosome slippage) involved:
0 The ribosome read-through involved:
no

The alternative forms of this protein occur by the alternative initiation of translation:

not tested

The ORF absolute position (the base range includes START and STOP codons or their equivalents):

189-3332

ORF

ORF position:

2

Version: `0`
Originaly submitted by: Václav Vopálenský	Reviewed by: Václav Vopálenský

The abbreviated name of this ORF/gene:
CAT

The description of the protein encoded in this ORF:
chloramphenicol acetyl transferase

The translational frameshift (ribosome slippage) involved:
0 The ribosome read-through involved:
no

The alternative forms of this protein occur by the alternative initiation of translation:

not tested

The ORF absolute position (the base range includes START and STOP codons or their equivalents):

4739-5398

Citations:

Holcik M., Lefebvre C., Yeh C., Chow T., Korneluk R. G. (1999) A new internal-ribosome-entry-site motif potentiates XIAP-mediated cytoprotection. Nat. Cell. Biol. 1(3):190-192

Baird S. D., Lewis S. M., Turcotte M., Holcik M. (2007) A search for structurally similar cellular internal ribosome entry sites. Nucleic. Acids. Res. 35(14):4664-4677

IRESs:

IRES:

Version: `2`	Last change: `2008-07-14 13:30:34`
Originaly submitted by: Václav Vopálenský	Reviewed by: Václav Vopálenský
The IRES name: `XIAP`	The functional status of IRES: `functional`

The IRES absolute position (the range includes START and STOP codons or their equivalents):

3621-3782

How IRES boundaries were determined:
experimentally_determined

5'-end of IRES relative to last base of the STOP codon of the upstream ORF: `289`	3'-end of IRES relative to last base of the STOP codon of the upstream ORF: `450`
5'-end of IRES relative to first base of the START codon of the downstream ORF: `-1118`	3'-end of IRES relative to first base of the START codon of the downstream ORF: `-957`

The sequence of IRES region aligned to its secondary structure (if available):

ATTAGAATGT TTCTTAGCGG TCGTGTAGTT ATTTTTATGT CATAAGTGGA TAATTTGTTA GCTCCTATAA CAAAAGTCTG TTGCTTGTGT TTCACATTTT 
((..(((... .......... ....((((.. ....)))).. .((((((((( (..((((((. ........)) )))).)))). ..)))))).. )))..))))) 
^3621      ^3631      ^3641      ^3651      ^3661      ^3671      ^3681      ^3691      ^3701      ^3711      

GGATTTCCTA ATATAATGTT CTCTTTTTAG AAAAGGTGGA CAAGTCCTAT TTTCAAGAGA AG
..))...... ........(( ((((...... ...(((.... .....))).. .....))))) ).
^3721      ^3731      ^3741      ^3751      ^3761      ^3771      ^3781

There is no Vienna RNA package installed on the server or some error/warning messages were output. Due to that maybe we cannot prepare 2D structures for display. The error/warning message was:

((..(((.................((((......))))...((((((((((..((((((.........)))))).))))...))))))..)))..)))))..))..............((((((.........(((.........))).......)))))).
ERROR: unbalanced brackets in make_pair_table

STDOUT was:

Remarks:

XIAP IRES represents part of XIAP 5' UTR (nt from -162 to nt -1 of the original sequence). This region
retained full IRES activity comparable with the whole 5' UTR (from nt -1007 to nt -1) of XIAP mRNA.

Citations:

Holcik M., Lefebvre C., Yeh C., Chow T., Korneluk R. G. (1999) A new internal-ribosome-entry-site motif potentiates XIAP-mediated cytoprotection. Nat. Cell. Biol. 1(3):190-192

The translation experiments:

Translation results:

IRESite Translation Id: 537

Version: `2`	Last change: `2008-07-13 10:56:51`
Originaly submitted by: Václav Vopálenský	Reviewed by: Václav Vopálenský

The translation method used to study IRES function:
in vivo

The organism used for translation:

Homo sapiens HeLa (ATCC CCL-2)

The temperature (in degrees of Celsia):
37

The relative translation efficiency in % of this IRES:

100.000

	Name of the plasmid used as the negative control. `pbetaGAL/CAT`
	`IRESite Id` of the plasmid used as negative control. `237`
	The relative translation efficiency in % of the negative control: `1.000`
	The size (length) of intercistronic region in the negative control: `336`

The effect of 5'-cap analogs on translation:
not tested

Rapamycin affects translation:
not tested

Type of RNA subject to translation:

endogenous_nuclear_RNA_Pol_II_transcript

Remarks:

Translational data are derived from Fig. 1B.

Citations:

Holcik M., Lefebvre C., Yeh C., Chow T., Korneluk R. G. (1999) A new internal-ribosome-entry-site motif potentiates XIAP-mediated cytoprotection. Nat. Cell. Biol. 1(3):190-192

Translation results:

IRESite Translation Id: 538

Version: `2`	Last change: `2009-09-03 19:21:43`
Originaly submitted by: Václav Vopálenský	Reviewed by: Václav Vopálenský

The translation method used to study IRES function:
in vivo

The organism used for translation:

Homo sapiens HEK293 (ATCC CRL-1573)

The temperature (in degrees of Celsia):
37

The relative translation efficiency in % of this IRES:

100.000

	Name of the plasmid used as the negative control. `pBiCmod`
	`IRESite Id` of the plasmid used as negative control. `636`
	The relative translation efficiency in % of the negative control: `0.300`
	The size (length) of intercistronic region in the negative control: `294`

The effect of 5'-cap analogs on translation:
not tested

Rapamycin affects translation:
not tested

Type of RNA subject to translation:

endogenous_nuclear_RNA_Pol_II_transcript

Remarks:

Translational data are derived from Fig. 4A.

Citations:

Baird S. D., Lewis S. M., Turcotte M., Holcik M. (2007) A search for structurally similar cellular internal ribosome entry sites. Nucleic. Acids. Res. 35(14):4664-4677

IRES trans-acting factor (ITAFS):

IRES trans-acting factor (ITAF):

Version: `0`
Originaly submitted by: Václav Vopálenský	Reviewed by: Václav Vopálenský

Type of the interaction between ITAF and the RNA subject to translation:
direct_interaction_with_rna

ITAF protein characteristics:

Version: `2`	Last change: `2009-08-29 12:19:15`
Originaly submitted by: Martin Mokrejš	Reviewed by: Martin Mokrejš

ITAF abbreviated name:
La

ITAF fullname:
La autoantigen

ITAF description (long):
La autoantigen (p52), 52 kDa RNA binding protein, predominantly localized to nucleus, unwinds the dsRNA in ATP-dependent manner, forms a dimer

3.1.2. Organisms or in vitro systems where this ITAF was functionally studied:

Organism or in vitro system where ITAF was shown:

Necessity of ITAF for translation in this particular organism or system:
stimulatory

Method used to demonstrate ITAF effect:
in_vitro In vitro system used to demonstrate ITAF effect:
other

Remarks:

HEK 293T cell extracts were used to demonstrate in vitro binding to XIAP RNA. Data from Figure 1A.

Citations:

Lewis S. M., Veyrier A., Hosszu Ungureanu N., Bonnal S., Vagner S., Holcik M. (2007) Subcellular relocalization of a trans-acting factor regulates XIAP IRES-dependent translation. Mol. Biol. Cell. 18(4):1302-1311

IRES trans-acting factor (ITAF):

Version: `0`
Originaly submitted by: Václav Vopálenský	Reviewed by: Václav Vopálenský

Type of the interaction between ITAF and the RNA subject to translation:
direct_interaction_with_rna

OPTIONAL: The interacting RNA base range (if any):
405-432

ITAF protein characteristics:

Version: `1`	Last change: `2008-06-27 20:14:51`
Originaly submitted by: Martin Mokrejš	Reviewed by: Martin Mokrejš

ITAF abbreviated name:
hnRNP_A1

ITAF fullname:
heterogeneous nuclear ribonucleoprotein A1

ITAF description (long):
heterogeneous nuclear ribonucleoprotein A1 (p37), 37 kDa

3.2.2. Organisms or in vitro systems where this ITAF was functionally studied:

Organism or in vitro system where ITAF was shown:

Necessity of ITAF for translation in this particular organism or system:
inhibitory

Method used to demonstrate ITAF effect:
both In vitro system used to demonstrate ITAF effect:
other

The organism where action of this ITAF was studied:

Homo sapiens HEK 293T/17 (ATCC CRL-11268)

Remarks:

HEK 293T cell extracts were used to demonstrate in vitro binding to XIAP RNA core RNP-binding sequence. Data
from Figures 1A and 2D.

Citations:

IRES trans-acting factor (ITAF):

Version: `0`
Originaly submitted by: Václav Vopálenský	Reviewed by: Václav Vopálenský

Type of the interaction between ITAF and the RNA subject to translation:
direct_interaction_with_rna

OPTIONAL: The interacting RNA base range (if any):
332-353

ITAF protein characteristics:

Version: `0`
Originaly submitted by: Martin Mokrejš	Reviewed by: Martin Mokrejš

ITAF abbreviated name:
hnRNP_C1/C2

ITAF fullname:
heterogeneous nuclear ribonucleoprotein C1/C2

ITAF description (long):
heterogeneous nuclear ribonucleoprotein C1/C2 (p44), 44 kDa

3.3.2. Organisms or in vitro systems where this ITAF was functionally studied:

Organism or in vitro system where ITAF was shown:

Necessity of ITAF for translation in this particular organism or system:
required_but_available_internally

Method used to demonstrate ITAF effect:
in_vitro In vitro system used to demonstrate ITAF effect:
other

Remarks:

HEK 293T cell extracts were used to demonstrate in vitro binding to XIAP RNA. Data from Lewis et al. (2007)
Fig. 1A and Lewis et al. (2005), Fig. 2.

Citations:

Lewis S. M., Holcik M. (2005) IRES in distress: translational regulation of the inhibitor of apoptosis proteins XIAP and HIAP2 during cell stress. Cell. Death. Differ. 12(6):547-553

IRES trans-acting factor (ITAF):

Version: `0`
Originaly submitted by: Václav Vopálenský	Reviewed by: Václav Vopálenský

Type of the interaction between ITAF and the RNA subject to translation:
direct_interaction_with_rna

ITAF protein characteristics:

Version: `0`
Originaly submitted by: Martin Mokrejš	Reviewed by: Václav Vopálenský

ITAF abbreviated name:
PTB-1

ITAF fullname:
polypyrimidine tract-binding protein isoform 1

ITAF description (long):
polypyrimidine tract-binding protein isoform 1 binds dsRNA with (CCU)n motif where n is at least 3. By SELEX approach it was found it binds to 5'-CAGCCUGGUGCCUCUCUUUCGG-3' (Singh et al. (1995) Science 268:1173-1176) but also UCUU or UCUUC within pyrimidine-rich sequence (Perez et al. (1997) RNA3:1334-1347).

3.4.2. Organisms or in vitro systems where this ITAF was functionally studied:

Organism or in vitro system where ITAF was shown:

Necessity of ITAF for translation in this particular organism or system:
required_but_available_internally

Method used to demonstrate ITAF effect:
both In vitro system used to demonstrate ITAF effect:
other

The organism where action of this ITAF was studied:

Homo sapiens HEK 293T/17 (ATCC CRL-11268)

Remarks:

Deletion of the XIAP 5' UTR polypyrimidine tract causes a loss of PTB binding and a complete loss of IRES
activity, but does not disrupt the binding of other ITAFs.

Overexpression of PTB represses XIAP IRES activity.

HEK 293T cell extracts were used to demonstrate in vitro binding to XIAP RNA. Data from Figure 8A/B/C.

Citations:

Baird S. D., Lewis S. M., Turcotte M., Holcik M. (2007) A search for structurally similar cellular internal ribosome entry sites. Nucleic. Acids. Res. 35(14):4664-4677

Regions with experimentally determined secondary structures:

A region with the experimentally determined secondary structure:

IRESite 2D Struct Id: 23

Version: `1`	Last change: `2008-07-14 15:28:11`
Originaly submitted by: Václav Vopálenský	Reviewed by: Václav Vopálenský
The function of the 2D structure: `IRES`	The 2D structure causes frameshift: `no`

The absolute position of the experimentally mapped region (the range includes START and STOP codons or their equivalents):
3621-3782

The underlying nucleic acid sequence and structure of the mapped region:

ATTAGAATGT TTCTTAGCGG TCGTGTAGTT ATTTTTATGT CATAAGTGGA TAATTTGTTA GCTCCTATAA CAAAAGTCTG TTGCTTGTGT TTCACATTTT 
((..(((... .......... ....((((.. ....)))).. .((((((((( (..((((((. ........)) )))).)))). ..)))))).. )))..))))) 
^3621      ^3631      ^3641      ^3651      ^3661      ^3671      ^3681      ^3691      ^3701      ^3711      

GGATTTCCTA ATATAATGTT CTCTTTTTAG AAAAGGTGGA CAAGTCCTAT TTTCAAGAGA AG
..))...... ........(( ((((...... ...(((.... .....))).. .....))))) ).
^3721      ^3731      ^3741      ^3751      ^3761      ^3771      ^3781

There is no Vienna RNA package installed on the server or some error/warning messages were output. Due to that maybe we cannot prepare 2D structures for display. The error/warning message was:

((..(((.................((((......))))...((((((((((..((((((.........)))))).))))...))))))..)))..)))))..))..............((((((.........(((.........))).......)))))).
ERROR: unbalanced brackets in make_pair_table

STDOUT was:

Remarks:

2D structure of XIAP from Fig 1.

4.1.1. Enzymes used to characterize at least partially the 2D structure.