LOCUS Exported 14707 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gene targeting vector for the mouse Rosa26 locus, including a loxP flanked stop cassette and GFP, for gateway cloning. ACCESSION . VERSION . KEYWORDS pR26 GFP Dest SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 14707) AUTHORS Chu VT, Weber T, Graf R, Sommermann T, Petsch K, Sack U, Volchkov P, Rajewsky K, Kuhn R TITLE Efficient generation of Rosa26 knock-in mice using CRISPR/Cas9 in C57BL/6 zygotes. JOURNAL BMC Biotechnol. 2016 Jan 16;16(1):4. doi: 10.1186/s12896-016-0234-4. PUBMED 26772810 REFERENCE 2 (bases 1 to 14707) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..14707 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 5..1087 /label=Rosa26 left arm /note="left homology arm from intron 1 of the mouse Rosa26 gene (Soriano, 1999)" misc_feature 1094..1197 /label=Ad SA /note="adenovirus major late transcript splice acceptor (Friedrich and Soriano, 1991)" protein_bind 1208..1241 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." polyA_signal 1248..1296 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" CDS complement(1303..2106) /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(1414..1433) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 2024..2043 /label=Neo-R /note="Neomycin resistance gene, reverse primer" intron 2920..2985 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 3115..3135 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 3560..3694 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3597..3616) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3651..3670 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" protein_bind 3714..3747 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind 3760..3884 /gene="mutant version of attR" /label=attR1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" promoter 3909..3939 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" CDS 3993..4652 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(4059..4078) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" CDS 4994..5299 /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 5178..5197 /label=ccdB-fwd /note="ccdB gene, forward primer" protein_bind complement(5340..5464) /gene="mutant version of attR" /label=attR2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind 5499..5546 /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)." misc_feature 5585..6136 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(5729..5746) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 5956..5975 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 6148..6867 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 6148..6864 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(6193..6214) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(6454..6473) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 6801..6822 /label=EGFP-C /note="EGFP, forward primer" protein_bind 6892..6939 /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)." primer_bind complement(6964..6980) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6964..6980) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(6993..7010) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 6999..7223 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" misc_feature 7311..11651 /label=Rosa26 right arm /note="right homology arm from intron 1 of the mouse Rosa26 gene (Soriano, 1999)" primer_bind complement(11736..11759) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind complement(11862..11882) /label=T3 /note="T3 promoter, forward primer" promoter complement(11862..11880) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(11901..11917) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(11901..11917) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(11914..11936) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 11925..11941 /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(11949..11979) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 11994..12015 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(12132..12149) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(12303..12891) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(12383..12402) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(13062..13922) /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 13685..13704 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(13923..14027) /gene="bla" /label=AmpR promoter rep_origin complement(14053..14508) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(14190..14211) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 14402..14421 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 14635..14657 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 14649..14666 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 14650..14666 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 14676..14695 /label=T7 /note="T7 promoter, forward primer" promoter 14676..14694 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" ORIGIN 1 ccccgcggca ggccctccga gcgtggtgga gccgttctgt gagacagccg ggtacgagtc 61 gtgacgctgg aaggggcaag cgggtggtgg gcaggaatgc ggtccgccct gcagcaaccg 121 gagggggagg gagaagggag cggaaaagtc tccaccggac gcggccatgg ctcggggggg 181 ggggggcagc ggaggagcgc ttccggccga cgtctcgtcg ctgattggct tcttttcctc 241 ccgccgtgtg tgaaaacaca aatggcgtgt tttggttggc gtaaggcgcc tgtcagttaa 301 cggcagccgg agtgcgcagc cgccggcagc ctcgctctgc ccactgggtg gggcgggagg 361 taggtggggt gaggcgagct ggacgtgcgg gcgcggtcgg cctctggcgg ggcgggggag 421 gggagggagg gtcagcgaaa gtagctcgcg cgcgagcggc cgcccaccct ccccttcctc 481 tgggggagtc gttttacccg ccgccggccg ggcctcgtcg tctgattggc tctcggggcc 541 cagaaaactg gcccttgcca ttggctcgtg ttcgtgcaag ttgagtccat ccgccggcca 601 gcgggggcgg cgaggaggcg ctcccaggtt ccggccctcc cctcggcccc gcgccgcaga 661 gtctggccgc gcgcccctgc gcaacgtggc aggaagcgcg cgctgggggc ggggacgggc 721 agtagggctg agcggctgcg gggcgggtgc aagcacgttt ccgacttgag ttgcctcaag 781 aggggcgtgc tgagccagac ctccatcgcg cactccgggg agtggaggga aggagcgagg 841 gctcagttgg gctgttttgg aggcaggaag cacttgctct cccaaagtcg ctctgagttg 901 ttatcagtaa gggagctgca gtggagtagg cggggagaag gccgcaccct tctccggagg 961 ggggagggga gtgttgcaat acctttctgg gagttctctg ctgcctcctg gcttctgagg 1021 accgccctgg gcctgggaga atcccttccc cctcttccct cgtgatctgc aactccagtc 1081 tttctagcat ctgtagggcg cagtagtcca gggtttcctt gatgatgtca tacttatcct 1141 gtcccttttt tttccacagc tcgcggttga ggacaaactc ttcgcggtct ttccagtggt 1201 taattaaata acttcgtata gcatacatta tacgaagtta tggatccgaa caaacgaccc 1261 aacacccgtg cgttttattc tgtcttttta ttgccgatcc cctcagaaga actcgtcaag 1321 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa 1381 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc 1441 ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt 1501 ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc 1561 gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc 1621 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg 1681 atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat 1741 tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg 1801 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac 1861 agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag 1921 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga 1981 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa 2041 tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatggccga 2101 tcccatattg gctgcagggt cgctcggtgt tcgaggccac acgcgtcacc ttaatatgcg 2161 aagtggacct gggaccgcgc cgccccgact gcatctgcgt gttcgaattc gccaatgaca 2221 agacgctggg cggggtttgc tcgacattgg gtggaaacat tccaggcctg ggtggagagg 2281 ctttttgctt cctcttgcaa aaccacactg ctcgacattg ggtggaaaca ttccaggcct 2341 gggtggagag gctttttgct tcctcttgca aaaccacact gctcgactag tgattaaagt 2401 cgactcgggg acaccaaata tggcgatctc ggccttttcg tttcttggag ctgggacatg 2461 tttgccatcg atccatctac caccagaacg gccgttagat ctgctgccac cgttgtttcc 2521 accgaagaaa ccaccgttgc cgtaaccacc acgacggttg ttgctaaaga agctgccacc 2581 gccacggcca ccgttgtagc cgccgttgtt gttattgtag ttgctcatgt tatttctggc 2641 acttcttggt tttcctctta agtgaggagg aacataacca ttctcgttgt tgtcgttgat 2701 gcttaaattt tgcacttgtt cgctcagttc agccataata tgaaatgctt ttcttgttgt 2761 tcttacggaa taccacttgc cacctatcac cacaactaac tttttcccgt tcctccatct 2821 cttttatatt ttttttctcg agggatcttt gtgaaggaac cttacttctg tggtgtgaca 2881 taattggaca aactacctac agagatttaa agctctaagg taaatataaa atttttaagt 2941 gtataatgtg ttaaactact gattctaatt gtttgtgtat tttagattcc aacctatgga 3001 actgatgaat gggagcagtg gtggaatgcc tttaatgagg aaaacctgtt ttgctcagaa 3061 gaaatgccat ctagtgatga tgaggctact gctgactctc aacattctac tcctccaaaa 3121 aagaagagaa aggtagaaga ccccaaggac tttccttcag aattgctaag ttttttgagt 3181 catgctgtgt ttagtaatag aactcttgct tgctttgcta tttacaccac aaaggaaaaa 3241 gctgcactgc tatacaagaa aattatggaa aaatattctg taacctttat aagtaggcat 3301 aacagttata atcataacat actgtttttt cttactccac acaggcatag agtgtctgct 3361 attaataact atgctcaaaa attgtgtacc tttagctttt taatttgtaa aggggttaat 3421 aaggaatatt tgatgtatag tgccttgact agagatcata atcagccata ccacatttgt 3481 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat 3541 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa 3601 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 3661 caaactcatc aatgtatctt atcatgtctg gatctgacat ggtaagtaag cttataactt 3721 cgtatagcat acattatacg aagttatcgg cgcgccatca caagtttgta caaaaaagct 3781 gaacgagaaa cgtaaaatga tataaatatc aatatattaa attagatttt gcataaaaaa 3841 cagactacat aatactgtaa aacacaacat atccagtcat attggcggcc gcattaggca 3901 ccccaggctt tacactttat gcttccggct cgtataatgt gtggattttg agttaggatc 3961 cgtcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca 4021 ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc 4081 aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga 4141 aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc 4201 atccggaatt ccgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc 4261 cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc 4321 acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa 4381 acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct 4441 gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg 4501 ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc 4561 aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac 4621 agtactgcga tgagtggcag ggcggggcgt aaacgcgtgg atccggctta ctaaaagcca 4681 gataacagta tgcgtatttg cgcgctgatt tttgcggtat aagaatatat actgatatgt 4741 atacccgaag tatgtcaaaa agaggtatgc tatgaagcag cgtattacag tgacagttga 4801 cagcgacagc tatcagttgc tcaaggcata tatgatgtca atatctccgg tctggtaagc 4861 acaaccatgc agaatgaagc ccgtcgtctg cgtgccgaac gctggaaagc ggaaaatcag 4921 gaagggatgg ctgaggtcgc ccggtttatt gaaatgaacg gctcttttgc tgacgagaac 4981 aggggctggt gaaatgcagt ttaaggttta cacctataaa agagagagcc gttatcgtct 5041 gtttgtggat gtacagagtg atattattga cacgcccggg cgacggatgg tgatccccct 5101 ggccagtgca cgtctgctgt cagataaagt ctcccgtgaa ctttacccgg tggtgcatat 5161 cggggatgaa agctggcgca tgatgaccac cgatatggcc agtgtgccgg tctccgttat 5221 cggggaagaa gtggctgatc tcagccaccg cgaaaatgac atcaaaaacg ccattaacct 5281 gatgttctgg ggaatataaa tgtcaggctc ccttatacac agccagtctg caggtcgacc 5341 atagtgactg gatatgttgt gttttacagc attatgtagt ctgtttttta tgcaaaatct 5401 aatttaatat attgatattt atatcatttt acgtttctcg ttcagctttc ttgtacaaag 5461 tggtgatggc gcgccgcggc cgcgatcaat tcggtaccga agttcctatt ccgaagttcc 5521 tattctctag aaagtatagg aacttcctcg agggtacccc aattccgccc cccccccccc 5581 cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta 5641 ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc 5701 ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat 5761 gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc 5821 ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt 5881 gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt 5941 gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag 6001 aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt 6061 tagtcgaggt taaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa 6121 aaacacgatg ataatatggc cacaaccatg gtgagcaagg gcgaggagct gttcaccggg 6181 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc 6241 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc 6301 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc 6361 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa 6421 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc 6481 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc 6541 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc 6601 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac 6661 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac 6721 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac 6781 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact 6841 ctcggcatgg acgagctgta caagtaatcc gggatccgga gagctcccaa cgaagttcct 6901 attccgaagt tcctattctc tagaaagtat aggaacttcc tcgaggttgg atgcagcccg 6961 ggggatccac tagttctaga gcggccgatc agcctcgact gtgccttcta gttgccagcc 7021 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt 7081 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct 7141 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc 7201 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctcgagatc 7261 cactagttct agcctcgagg ctagagcggc cgccgcggat atcgaattcg ctagaagatg 7321 ggcgggagtc ttctgggcag gcttaaaggc taacctggtg tgtgggcgtt gtcctgcagg 7381 ggaattgaac aggtgtaaaa ttggagggac aagacttccc acagattttc ggttttgtcg 7441 ggaagttttt taataggggc aaataaggaa aatgggagga taggtagtca tctggggttt 7501 tatgcagcaa aactacaggt tattattgct tgtgatccgc ctcggagtat tttccatcga 7561 ggtagattaa agacatgctc acccgagttt tatactctcc tgcttgagat ccttactaca 7621 gtatgaaatt acagtgtcgc gagttagact atgtaagcag aattttaatc atttttaaag 7681 agcccagtac ttcatatcca tttctcccgc tccttctgca gccttatcaa aaggtatttt 7741 agaacactca ttttagcccc attttcattt attatactgg cttatccaac ccctagacag 7801 agcattggca ttttcccttt cctgatctta gaagtctgat gactcatgaa accagacaga 7861 ttagttacat acaccacaaa tcgaggctgt agctggggcc tcaacactgc agttctttta 7921 taactcctta gtacactttt tgttgatcct ttgccttgat ccttaatttt cagtgtctat 7981 cacctctccc gtcaggtggt gttccacatt tgggcctatt ctcagtccag ggagttttac 8041 aacaatagat gtattgagaa tccaacctaa agcttaactt tccactccca tgaatgcctc 8101 tctccttttt ctccatttat aaactgagct attaaccatt aatggtttcc aggtggatgt 8161 ctcctccccc aatattacct gatgtatctt acatattgcc aggctgatat tttaagacat 8221 taaaaggtat atttcattat tgagccacat ggtattgatt actgcttact aaaattttgt 8281 cattgtacac atctgtaaaa ggtggttcct tttggaatgc aaagttcagg tgtttgttgt 8341 ctttcctgac ctaaggtctt gtgagcttgt attttttcta tttaagcagt gctttctctt 8401 ggactggctt gactcatggc attctacacg ttattgctgg tctaaatgtg attttgccaa 8461 gcttcttcag gacctataat tttgcttgac ttgtagccaa acacaagtaa aatgattaag 8521 caacaaatgt atttgtgaag cttggttttt aggttgttgt gttgtgtgtg cttgtgctct 8581 ataataatac tatccagggg ctggagaggt ggctcggagt tcaagagcac agactgctct 8641 tccagaagtc ctgagttcaa ttcccagcaa ccacatggtg gctcacaacc atctgtaatg 8701 ggatctgatg ccctcttctg gtgtgtctga agaccacaag tgtattcaca ttaaataaat 8761 aaatcctcct tcttcttctt tttttttttt ttaaagagaa tactgtctcc agtagaattt 8821 actgaagtaa tgaaatactt tgtgtttgtt ccaatatggt agccaataat caaattactc 8881 tttaagcact ggaaatgtta ccaaggaact aatttttatt tgaagtgtaa ctgtggacag 8941 aggagccata actgcagact tgtgggatac agaagaccaa tgcagacttt aatgtctttt 9001 ctcttacact aagcaataaa gaaataaaaa ttgaacttct agtatcctat ttgtttaaac 9061 tgctagcttt acttaacttt tgtgcttcat ctatacaaag ctgaaagcta agtctgcagc 9121 cattactaaa catgaaagca agtaatgata attttggatt tcaaaaatgt agggccagag 9181 tttagccagc cagtggtggt gcttgccttt atgcctttaa tcccagcact ctggaggcag 9241 agacaggcag atctctgagt ttgagcccag cctggtctac acatcaagtt ctatctagga 9301 tagccaggaa tacacacaga aaccctgttg gggagggggg ctctgagatt tcataaaatt 9361 ataattgaag cattccctaa tgagccacta tggatgtggc taaatccgtc tacctttctg 9421 atgagatttg ggtattattt tttctgtctc tgctgttggt tgggtctttt gacactgtgg 9481 gctttcttta aagcctcctt cctgccatgt ggtctcttgt ttgctactaa cttcccatgg 9541 cttaaatggc atggcttttt gccttctaag ggcagctgct gagatttgca gcctgatttc 9601 cagggtgggg ttgggaaatc tttcaaacac taaaattgtc ctttaatttt ttttttaaaa 9661 aatgggttat ataataaacc tcataaaata gttatgagga gtgaggtgga ctaatattaa 9721 atgagtccct cccctataaa agagctatta aggctttttg tcttatactt aacttttttt 9781 ttaaatgtgg tatctttaga accaagggtc ttagagtttt agtatacaga aactgttgca 9841 tcgcttaatc agattttcta gtttcaaatc cagagaatcc aaattcttca cagccaaagt 9901 caaattaaga atttctgact tttaatgtta atttgcttac tgtgaatata aaaatgatag 9961 cttttcctga ggcagggtct cactatgtat ctctgcctga tctgcaacaa gatatgtaga 10021 ctaaagttct gcctgctttt gtctcctgaa tactaaggtt aaaatgtagt aatacttttg 10081 gaacttgcag gtcagattct tttatagggg acacactaag ggagcttggg tgatagttgg 10141 taaaatgtgt ttcaagtgat gaaaacttga attattatca ccgcaaccta ctttttaaaa 10201 aaaaaagcca ggcctgttag agcatgctta agggatccct aggacttgct gagcacacaa 10261 gagtagttac ttggcaggct cctggtgaga gcatatttca aaaaacaagg cagacaacca 10321 agaaactaca gttaaggtta cctgtcttta aaccatctgc atatacacag ggatattaaa 10381 atattccaaa taatatttca ttcaagtttt cccccatcaa attgggacat ggatttctcc 10441 ggtgaatagg cagagttgga aactaaacaa atgttggttt tgtgatttgt gaaattgttt 10501 tcaagtgata gttaaagccc atgagataca gaacaaagct gctatttcga ggtctcttgg 10561 tttatactca gaagcacttc tttgggtttc cctgcactat cctgatcatg tgctaggcct 10621 accttaggct gattgttgtt caaataaact taagtttcct gtcaggtgat gtcatatgat 10681 ttcatatatc aaggcaaaac atgttatata tgttaaacat ttgtacttaa tgtgaaagtt 10741 aggtctttgt gggtttgatt tttaattttc aaaacctgag ctaaataagt catttttaca 10801 tgtcttacat ttggtggaat tgtataattg tggtttgcag gcaagactct ctgacctagt 10861 aaccctacct atagagcact ttgctgggtc acaagtctag gagtcaagca tttcaccttg 10921 aagttgagac gttttgttag tgtatactag tttatatgtt ggaggacatg tttatccaga 10981 agatattcag gactattttt gactgggcta aggaattgat tctgattagc actgttagtg 11041 agcattgagt ggcctttagg cttgaattgg agtcacttgt atatctcaaa taatgctggc 11101 cttttttaaa aagcccttgt tctttatcac cctgttttct acataatttt tgttcaaaga 11161 aatacttgtt tggatctcct tttgacaaca atagcatgtt ttcaagccat attttttttc 11221 cttttttttt ttttttttgg tttttcgaga cagggtttct ctgtatagcc ctggctgtcc 11281 tggaactcac tttgtagacc aggctggcct cgaactcaga aatccgcctg cctctgcctc 11341 ctgagtgccg ggattaaagg cgtgcaccac cacgcctggc taagttggat attttgttat 11401 ataactataa ccaatactaa ctccactggg tggattttta attcagtcag tagtcttaag 11461 tggtctttat tggcccttca ttaaaatcta ctgttcactc taacagaggc tgttggtact 11521 agtggcactt aagcaacttc ctacggatat actagcagat taagggtcag ggatagaaac 11581 tagtctagcg ttttgtatac ctaccagctt tatactacct tgttctgata gaaatatttc 11641 aggacatcta gcttatcgat accgtcgacg gtatcgataa gcttgatatc gaattctacc 11701 gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg 11761 gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc 11821 gccgccgccc tgcaggcgat cgctacgtac agcttttgtt ccctttagtg agggttaatt 11881 gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca 11941 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg 12001 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 12061 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 12121 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 12181 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 12241 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 12301 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 12361 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 12421 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 12481 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 12541 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 12601 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 12661 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 12721 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt 12781 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 12841 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 12901 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 12961 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 13021 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 13081 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 13141 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 13201 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 13261 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 13321 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 13381 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 13441 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 13501 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 13561 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 13621 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 13681 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 13741 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 13801 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 13861 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 13921 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 13981 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 14041 aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt 14101 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag 14161 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga 14221 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg 14281 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc 14341 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg 14401 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc 14461 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat 14521 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc 14581 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt 14641 cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat 14701 tggagct //