LOCUS Exported 14015 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION piggyBac expression vector with TET-ON system and puromycin selection. ACCESSION . VERSION . KEYWORDS PB-TET-PH-lox66FRT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 14015) TITLE Minoru Ko Lab plasmids REFERENCE 2 (bases 1 to 14015) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..14015 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(1..456) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(138..159) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 350..369 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 598..614 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 621..640 /label=T7 /note="T7 promoter, forward primer" promoter 621..639 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" repeat_region 706..768 /label=piggyBac right (3') inverted repeat /note="piggyBac transposon-specific inverted terminal repeat sequence (ITR)" primer_bind 1177..1193 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 1204..1703 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(1239..1262) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 1636..1655 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 1727..2323 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(1727..1746) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 2223..2243 /label=Puro-F /note="Puromycin resistance gene, forward primer" CDS 2330..3325 /codon_start=1 /product="truncated herpes simplex virus thymidine kinase (Salomon et al., 1995)" /label=-Delta-TK /note="confers sensitivity to nucleoside analogs such as acyclovir or ganciclovir" /translation="MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRV LGASETIANIYTTQHRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGS SHAPPPALTLIFDRHPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGAL PEDRHIDRLAKRQRPGERLDLAMLAAIRRVYGLLANTVRYLQCGGSWREDWGQLSGTAV PPQGAEPQSNAGPRPHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRSMHVFI LDYDQSPAGCRDALLQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN" primer_bind complement(3959..3975) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 3983..3999 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(4007..4037) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4052..4073 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." protein_bind 4156..4174 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 4193..4211 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 4230..4248 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 4267..4285 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 4304..4322 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 4341..4359 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 4378..4396 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 4415..4433 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 4455..4479 /label=LNCX /note="Human CMV promoter, forward primer" protein_bind 4559..4683 /gene="mutant version of attR" /label=attR1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" promoter 4708..4738 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" CDS 4792..5451 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(4858..4877) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" CDS 5793..6098 /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind 5977..5996 /label=ccdB-fwd /note="ccdB gene, forward primer" protein_bind complement(6139..6263) /gene="mutant version of attR" /label=attR2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" misc_feature 6315..6888 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" misc_feature 6338..6890 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(6482..6499) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 6709..6728 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 6929..9970 /codon_start=1 /gene="lacZ" /product="beta-galactosidase" /label=lacZ /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD SWSPSVSAEFQLSAGRYHYQLVWCQ" CDS 9995..10798 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(10058..10077) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 10668..10687 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(10830..10847) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 10836..11060 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 11499..11533 /label=piggyBac left (5') inverted repeat /note="piggyBac transposon-specific inverted terminal repeat sequence (ITR)" protein_bind 11560..11593 /label=lox66 /bound_moiety="Cre recombinase" /note="Right element (RE) mutant of loxP (Araki et al., 2010). Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind 11626..11673 /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." promoter complement(11825..11843) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(11864..11880) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 11888..11904 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(11912..11942) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 11957..11978 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(12095..12112) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(12266..12854) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(12346..12365) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(13025..13885) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 13648..13667 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(13886..13990) /gene="bla" /label=AmpR promoter ORIGIN 1 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat 61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga 121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca 181 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct 241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc 301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag 361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca 421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca 481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg 541 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta 601 aaacgacggc cagtgaattg taatacgact cactataggg cgaattggag ctcggtattc 661 acgacagcag gctgaataat aaaaaaatta gaaactatta tttaacccta gaaagataat 721 catattgtga cgtacgttaa agataatcat gcgtaaaatt gacgcatgtg ttttatcggt 781 ctgtatatcg aggtttattt attaatttga atagatatta agttttatta tatttacact 841 tacatactaa taataaattc aacaaacaat ttatttatgt ttatttattt attaaaaaaa 901 aacaaaaact caaaatttct tctataaagt aacaaaactt ttaaacattc tctcttttac 961 aaaaataaac ttattttgta ctttaaaaac agtcatgttg tattataaaa taagtaatta 1021 gcttaaccta tacataatag aaacaaatta tacttattag tcagtcagaa acaactttgg 1081 cacatatcaa tattatgctc tcgttactgg cgaaaggggg atgtgctgca aggcgattaa 1141 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattct 1201 accgggtagg ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc 1261 tgggcacttg gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta 1321 ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag 1381 tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag 1441 cacgtctcac tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc 1501 tttggggcag cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa 1561 ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt 1621 cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct 1681 cttcctcatc tccgggcctt tcgaccgatc atcaagcttg atcctcatga ccgagtacaa 1741 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc 1801 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg 1861 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg 1921 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg 1981 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc 2041 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct 2101 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct 2161 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc 2221 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc 2281 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gccggatcca tgcccacgct 2341 actgcgggtt tatatagacg gtccccacgg gatggggaaa accaccacca cgcaactgct 2401 ggtggccctg ggttcgcgcg acgatatcgt ctacgtaccc gagccgatga cttactggcg 2461 ggtgctgggg gcttccgaga caatcgcgaa catctacacc acacaacacc gcctcgacca 2521 gggtgagata tcggccgggg acgcggcggt ggtaatgaca agcgcccaga taacaatggg 2581 catgccttat gccgtgaccg acgccgttct ggctcctcat atcggggggg aggctgggag 2641 ctcacatgcc ccgcccccgg ccctcaccct catcttcgac cgccatccca tcgccgccct 2701 cctgtgctac ccggccgcgc ggtaccttat gggcagcatg accccccagg ccgtgctggc 2761 gttcgtggcc ctcatcccgc cgaccttgcc cggcaccaac atcgtgcttg gggcccttcc 2821 ggaggacaga cacatcgacc gcctggccaa acgccagcgc cccggcgagc ggctggacct 2881 ggctatgctg gctgcgattc gccgcgttta cgggctactt gccaatacgg tgcggtatct 2941 gcagtgcggc gggtcgtggc gggaggactg gggacagctt tcggggacgg ccgtgccgcc 3001 ccagggtgcc gagccccaga gcaacgcggg cccacgaccc catatcgggg acacgttatt 3061 taccctgttt cgggcccccg agttgctggc ccccaacggc gacctgtata acgtgtttgc 3121 ctgggccttg gacgtcttgg ccaaacgcct ccgttccatg cacgtcttta tcctggatta 3181 cgaccaatcg cccgccggct gccgggacgc cctgctgcaa cttacctccg ggatggtcca 3241 gacccacgtc accacccccg gctccatacc gacgatatgc gacctggcgc gcacgtttgc 3301 ccgggagatg ggggaggcta actgagctct agcttatcga ctcgccgata gtggaaaccg 3361 acgccccagc actcgtccga gggcaaagga atagtcgaga aattgatgat ctattaaaca 3421 ataaagatgt ccacatggaa gtttttcctg tcatactttg ttaagaaggg tgagaacaga 3481 gtacctacat tttgaatgga aggattggag ctacgggggt gggggtgggg tgggattaga 3541 taaatgcctg ctctttactg aaggctcttt actattgctt tatgataatg tttcatagtt 3601 ggatatcata atttaaacaa gcaaaaccaa attaagggcc agctcattcc tcccactcat 3661 gatctataga tcctctagag agctcacggg gacagccccc ccccaaagcc cccagggatg 3721 taattacgtc cctcccccgc tagggggcag cagcgagccg cccggggctc cgctccggtc 3781 cggcgctccc cccgcatccc cgagccggca gcgtgcgggg acagcccggg cacggggaag 3841 gtggcacggg atcgctttcc tctgaacgct tctcgctgct ctttgagcct gcagacacct 3901 gggggatacg gggaaaaagc ttgtcgacct gcaggcatgc aagcttggcg taatcatggt 3961 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 4021 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 4081 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagtaactc gagggatcca 4141 agctggccgc acgtctccct atcagtgata gagaagtcga cacgtctcga gctccctatc 4201 agtgatagag aaggtacgtc tagaacgtct ccctatcagt gatagagaag tcgacacgtc 4261 tcgagctccc tatcagtgat agagaaggta cgtctagaac gtctccctat cagtgataga 4321 gaagtcgaca cgtctcgagc tccctatcag tgatagagaa ggtacgtcta gaacgtctcc 4381 ctatcagtga tagagaagtc gacacgtctc gagctcccta tcagtgatag agaaggtacc 4441 ccctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc 4501 tgttttgacc tccatagaag acaccgggac cgatccagcc tccgcggtgg cggccatcac 4561 aagtttgtac aaaaaagctg aacgagaaac gtaaaatgat ataaatatca atatattaaa 4621 ttagattttg cataaaaaac agactacata atactgtaaa acacaacata tccagtcact 4681 atggcggccg cattaggcac cccaggcttt acactttatg cttccggctc gtataatgtg 4741 tggattttga gttaggatcc gtcgagattt tcaggagcta aggaagctaa aatggagaaa 4801 aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag 4861 gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc 4921 tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt 4981 gcccgcctga tgaatgctca tccggaattc cgtatggcaa tgaaagacgg tgagctggtg 5041 atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca 5101 tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat 5161 gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt 5221 ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg 5281 gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg 5341 ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga 5401 atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta aacgcgtgga 5461 tccggcttac taaaagccag ataacagtat gcgtatttgc gcgctgattt ttgcggtata 5521 agaatatata ctgatatgta tacccgaagt atgtcaaaaa gaggtatgct atgaagcagc 5581 gtattacagt gacagttgac agcgacagct atcagttgct caaggcatat atgatgtcaa 5641 tatctccggt ctggtaagca caaccatgca gaatgaagcc cgtcgtctgc gtgccgaacg 5701 ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc cggtttattg aaatgaacgg 5761 ctcttttgct gacgagaaca ggggctggtg aaatgcagtt taaggtttac acctataaaa 5821 gagagagccg ttatcgtctg tttgtggatg tacagagtga tattattgac acgcccgggc 5881 gacggatggt gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac 5941 tttacccggt ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca 6001 gtgtgccggt ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca 6061 tcaaaaacgc cattaacctg atgttctggg gaatataaat gtcaggctcc cttatacaca 6121 gccagtctgc aggtcgacca tagtgactgg atatgttgtg ttttacagta ttatgtagtc 6181 tgttttttat gcaaaatcta atttaatata ttgatattta tatcatttta cgtttctcgt 6241 tcagctttct tgtacaaagt ggtgatggcc gctctagcta gtcgataagc taattccgcc 6301 cccccccccc cccccccctc tccctccccc ccccctaacg ttactggccg aagccgcttg 6361 gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc 6421 aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc 6481 cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa 6541 gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct 6601 ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca 6661 caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca 6721 agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat 6781 ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc 6841 cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata tggccatgag 6901 cttgatgatc tgtgacatgg cggatcccgt cgttttacaa cgtcgtgact gggaaaaccc 6961 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag 7021 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg 7081 ctttgcctgg tttccggcac cagaagcggt gccggaaagc tggctggagt gcgatcttcc 7141 tgaggccgat actgtcgtcg tcccctcaaa ctggcagatg cacggttacg atgcgcccat 7201 ctacaccaac gtgacctatc ccattacggt caatccgccg tttgttccca cggagaatcc 7261 gacgggttgt tactcgctca catttaatgt tgatgaaagc tggctacagg aaggccagac 7321 gcgaattatt tttgatggcg ttaactcggc gtttcatctg tggtgcaacg ggcgctgggt 7381 cggttacggc caggacagtc gtttgccgtc tgaatttgac ctgagcgcat ttttacgcgc 7441 cggagaaaac cgcctcgcgg tgatggtgct gcgctggagt gacggcagtt atctggaaga 7501 tcaggatatg tggcggatga gcggcatttt ccgtgacgtc tcgttgctgc ataaaccgac 7561 tacacaaatc agcgatttcc atgttgccac tcgctttaat gatgatttca gccgcgctgt 7621 actggaggct gaagttcaga tgtgcggcga gttgcgtgac tacctacggg taacagtttc 7681 tttatggcag ggtgaaacgc aggtcgccag cggcaccgcg cctttcggcg gtgaaattat 7741 cgatgagcgt ggtggttatg ccgatcgcgt cacactacgt ctgaacgtcg aaaacccgaa 7801 actgtggagc gccgaaatcc cgaatctcta tcgtgcggtg gttgaactgc acaccgccga 7861 cggcacgctg attgaagcag aagcctgcga tgtcggtttc cgcgaggtgc ggattgaaaa 7921 tggtctgctg ctgctgaacg gcaagccgtt gctgattcga ggcgttaacc gtcacgagca 7981 tcatcctctg catggtcagg tcatggatga gcagacgatg gtgcaggata tcctgctgat 8041 gaagcagaac aactttaacg ccgtgcgctg ttcgcattat ccgaaccatc cgctgtggta 8101 cacgctgtgc gaccgctacg gcctgtatgt ggtggatgaa gccaatattg aaacccacgg 8161 catggtgcca atgaatcgtc tgaccgatga tccgcgctgg ctaccggcga tgagcgaacg 8221 cgtaacgcga atggtgcagc gcgatcgtaa tcacccgagt gtgatcatct ggtcgctggg 8281 gaatgaatca ggccacggcg ctaatcacga cgcgctgtat cgctggatca aatctgtcga 8341 tccttcccgc ccggtgcagt atgaaggcgg cggagccgac accacggcca ccgatattat 8401 ttgcccgatg tacgcgcgcg tggatgaaga ccagcccttc ccggctgtgc cgaaatggtc 8461 catcaaaaaa tggctttcgc tacctggaga gacgcgcccg ctgatccttt gcgaatacgc 8521 ccacgcgatg ggtaacagtc ttggcggttt cgctaaatac tggcaggcgt ttcgtcagta 8581 tccccgttta cagggcggct tcgtctggga ctgggtggat cagtcgctga ttaaatatga 8641 tgaaaacggc aacccgtggt cggcttacgg cggtgatttt ggcgatacgc cgaacgatcg 8701 ccagttctgt atgaacggtc tggtctttgc cgaccgcacg ccgcatccag cgctgacgga 8761 agcaaaacac cagcagcagt ttttccagtt ccgtttatcc gggcaaacca tcgaagtgac 8821 cagcgaatac ctgttccgtc atagcgataa cgagctcctg cactggatgg tggcgctgga 8881 tggtaagccg ctggcaagcg gtgaagtgcc tctggatgtc gctccacaag gtaaacagtt 8941 gattgaactg cctgaactac cgcagccgga gagcgccggg caactctggc tcacagtacg 9001 cgtagtgcaa ccgaacgcga ccgcatggtc agaagccggg cacatcagcg cctggcagca 9061 gtggcgtctg gcggaaaacc tcagtgtgac gctccccgcc gcgtcccacg ccatcccgca 9121 tctgaccacc agcgaaatgg atttttgcat cgagctgggt aataagcgtt ggcaatttaa 9181 ccgccagtca ggctttcttt cacagatgtg gattggcgat aaaaaacaac tgctgacgcc 9241 gctgcgcgat cagttcaccc gtgcaccgct ggataacgac attggcgtaa gtgaagcgac 9301 ccgcattgac cctaacgcct gggtcgaacg ctggaaggcg gcgggccatt accaggccga 9361 agcagcgttg ttgcagtgca cggcagatac acttgctgat gcggtgctga ttacgaccgc 9421 tcacgcgtgg cagcatcagg ggaaaacctt atttatcagc cggaaaacct accggattga 9481 tggtagtggt caaatggcga ttaccgttga tgttgaagtg gcgagcgata caccgcatcc 9541 ggcgcggatt ggcctgaact gccagctggc gcaggtagca gagcgggtaa actggctcgg 9601 attagggccg caagaaaact atcccgaccg ccttactgcc gcctgttttg accgctggga 9661 tctgccattg tcagacatgt ataccccgta cgtcttcccg agcgaaaacg gtctgcgctg 9721 cgggacgcgc gaattgaatt atggcccaca ccagtggcgc ggcgacttcc agttcaacat 9781 cagccgctac agtcaacagc aactgatgga aaccagccat cgccatctgc tgcacgcgga 9841 agaaggcaca tggctgaata tcgacggttt ccatatgggg attggtggcg acgactcctg 9901 gagcccgtca gtatcggcgg aattccagct gagcgccggt cgctaccatt accagttggt 9961 ctggtgtcag gggatccccc gggctgcagc caatatggga tcggccattg aacaagatgg 10021 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca 10081 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt 10141 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg 10201 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga 10261 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca 10321 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct 10381 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac 10441 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc 10501 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt 10561 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt 10621 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg 10681 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat 10741 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagg 10801 ggatcaattc tctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc 10861 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct 10921 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg 10981 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg 11041 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct cgatcctcta 11101 gagtcgaggg ctgcaggaat tcgatatcaa gctctagcca acaagctcgt catcgctttg 11161 cagaagagca gagaggatat gctcatcgtc taaagaacta cccattttat tatatattag 11221 tcacgatatc tataacaaga aaatatatat ataataagtt atcacgtaag tagaacatga 11281 aataacaata taattatcgt atgagttaaa tcttaaaagt cacgtaaaag ataatcatgc 11341 gtcattttga ctcacgcggt cgttatagtt caaaatcagt gacacttacc gcattgacaa 11401 gcacgcctca cgggagctcc aagcggcgac tgagatgtcc taaatgcaca gcgacggatt 11461 cgcgctattt agaaagagag agcaatattt caagaatgca tgcgtcaatt ttacgcagac 11521 tatctttcta gggttaaatt tgggcggtcg agggacctaa taacttcgta tagcatacat 11581 tatacgaacg gtaattaagg gttccggatc cccatcaagc taattgaagt tcctattccg 11641 aagttcctat tctctagaaa gtataggaac ttccagctgg aaatgatttg cgctttactc 11701 gacctaaact ttaaacacgt catagaatct tcgtttgaca aaaaccacat tgtggccaag 11761 ctgtgtgacg cgacgcgcgc taaagaatgg caaaccaagt cgcgcgaggt accagctttt 11821 gttcccttta gtgagggtta atttcgagct tggcgtaatc atggtcatag ctgtttcctg 11881 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 11941 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 12001 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 12061 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 12121 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 12181 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 12241 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 12301 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 12361 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 12421 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 12481 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 12541 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 12601 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 12661 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 12721 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 12781 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 12841 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 12901 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 12961 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 13021 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 13081 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 13141 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 13201 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 13261 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 13321 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 13381 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 13441 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 13501 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 13561 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 13621 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 13681 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 13741 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 13801 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 13861 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 13921 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 13981 gggttccgcg cacatttccc cgaaaagtgc cacct //