LOCUS Exported 16890 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gene targeting vector for the mouse Rosa26 locus, including a CAG promoter, loxP flanked stop cassette, GFP, for gateway cloning. ACCESSION . VERSION . KEYWORDS pR26 CAG/GFP Dest SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 16890) 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 16890) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..16890 /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)" polyA_signal 1438..1645 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" enhancer 1654..2033 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" intron 2309..3325 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 3249..3271 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 3333..3352 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" protein_bind 3388..3421 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." polyA_signal 3428..3476 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" CDS complement(3483..4286) /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(3594..3613) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 4204..4223 /label=Neo-R /note="Neomycin resistance gene, reverse primer" intron 5100..5165 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 5295..5315 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 5740..5874 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5777..5796) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5831..5850 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" protein_bind 5894..5927 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind 5940..6064 /gene="mutant version of attR" /label=attR1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" promoter 6089..6119 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" CDS 6173..6832 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(6239..6258) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" CDS 7174..7479 /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 7358..7377 /label=ccdB-fwd /note="ccdB gene, forward primer" protein_bind complement(7520..7644) /gene="mutant version of attR" /label=attR2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind 7679..7726 /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 7765..8316 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(7909..7926) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 8136..8155 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 8328..9047 /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 8328..9044 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(8373..8394) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(8634..8653) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 8981..9002 /label=EGFP-C /note="EGFP, forward primer" protein_bind 9072..9119 /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(9144..9160) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(9144..9160) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(9173..9190) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 9179..9403 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" misc_feature 9493..13833 /label=Rosa26 right arm /note="right homology arm from intron 1 of the mouse Rosa26 gene (Soriano, 1999)" primer_bind complement(13918..13941) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind complement(14045..14065) /label=T3 /note="T3 promoter, forward primer" promoter complement(14045..14063) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(14084..14100) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(14084..14100) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(14097..14119) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 14108..14124 /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(14132..14162) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 14177..14198 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(14315..14332) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(14486..15074) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(14566..14585) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(15245..16105) /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 15868..15887 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(16106..16210) /gene="bla" /label=AmpR promoter rep_origin complement(16236..16691) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(16373..16394) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 16585..16604 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 16818..16840 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 16832..16849 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 16833..16849 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 16859..16878 /label=T7 /note="T7 promoter, forward primer" promoter 16859..16877 /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 taattaactc ctttgagtgt gccttctagt tgccagccat ctgttgtttg cccctccccc 1261 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 1321 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 1381 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt ctagcgactg 1441 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg 1501 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga 1561 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg 1621 aagacaatag caggcatgct ggggatgcgg gtcgacattg attattgact agttattaat 1681 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac 1741 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa 1801 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt 1861 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc 1921 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat 1981 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg 2041 tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt 2101 atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg ggcgcgcgcc 2161 aggcggggcg gggcggggcg aggggcgggg cggggcgagg cggagaggtg cggcggcagc 2221 caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc 2281 ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc 2341 ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca 2401 ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg 2461 gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg 2521 cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg 2581 ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg 2641 cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg ggggctgcga 2701 ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc 2761 gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga gttgctgagc acggcccggc 2821 ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg 2881 gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg gctcggggga 2941 ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg 3001 ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga 3061 gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg 3121 cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct 3181 tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg 3241 cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt 3301 tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg tgctgtctca 3361 tcattttggc aaagaattct taattaaata acttcgtata gcatacatta tacgaagtta 3421 tggatccgaa caaacgaccc aacacccgtg cgttttattc tgtcttttta ttgccgatcc 3481 cctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg 3541 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca 3601 cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg 3661 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc 3721 acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc 3781 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga 3841 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca 3901 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg 3961 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct 4021 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc 4081 cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga 4141 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt 4201 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat 4261 ccatcttgtt caatggccga tcccatattg gctgcagggt cgctcggtgt tcgaggccac 4321 acgcgtcacc ttaatatgcg aagtggacct gggaccgcgc cgccccgact gcatctgcgt 4381 gttcgaattc gccaatgaca agacgctggg cggggtttgc tcgacattgg gtggaaacat 4441 tccaggcctg ggtggagagg ctttttgctt cctcttgcaa aaccacactg ctcgacattg 4501 ggtggaaaca ttccaggcct gggtggagag gctttttgct tcctcttgca aaaccacact 4561 gctcgactag tgattaaagt cgactcgggg acaccaaata tggcgatctc ggccttttcg 4621 tttcttggag ctgggacatg tttgccatcg atccatctac caccagaacg gccgttagat 4681 ctgctgccac cgttgtttcc accgaagaaa ccaccgttgc cgtaaccacc acgacggttg 4741 ttgctaaaga agctgccacc gccacggcca ccgttgtagc cgccgttgtt gttattgtag 4801 ttgctcatgt tatttctggc acttcttggt tttcctctta agtgaggagg aacataacca 4861 ttctcgttgt tgtcgttgat gcttaaattt tgcacttgtt cgctcagttc agccataata 4921 tgaaatgctt ttcttgttgt tcttacggaa taccacttgc cacctatcac cacaactaac 4981 tttttcccgt tcctccatct cttttatatt ttttttctcg agggatcttt gtgaaggaac 5041 cttacttctg tggtgtgaca taattggaca aactacctac agagatttaa agctctaagg 5101 taaatataaa atttttaagt gtataatgtg ttaaactact gattctaatt gtttgtgtat 5161 tttagattcc aacctatgga actgatgaat gggagcagtg gtggaatgcc tttaatgagg 5221 aaaacctgtt ttgctcagaa gaaatgccat ctagtgatga tgaggctact gctgactctc 5281 aacattctac tcctccaaaa aagaagagaa aggtagaaga ccccaaggac tttccttcag 5341 aattgctaag ttttttgagt catgctgtgt ttagtaatag aactcttgct tgctttgcta 5401 tttacaccac aaaggaaaaa gctgcactgc tatacaagaa aattatggaa aaatattctg 5461 taacctttat aagtaggcat aacagttata atcataacat actgtttttt cttactccac 5521 acaggcatag agtgtctgct attaataact atgctcaaaa attgtgtacc tttagctttt 5581 taatttgtaa aggggttaat aaggaatatt tgatgtatag tgccttgact agagatcata 5641 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc 5701 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat 5761 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 5821 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatctgacat 5881 ggtaagtaag cttataactt cgtatagcat acattatacg aagttatcgg cgcgccatca 5941 caagtttgta caaaaaagct gaacgagaaa cgtaaaatga tataaatatc aatatattaa 6001 attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat atccagtcac 6061 tatggcggcc gcattaggca ccccaggctt tacactttat gcttccggct cgtataatgt 6121 gtggattttg agttaggatc cgtcgagatt ttcaggagct aaggaagcta aaatggagaa 6181 aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga 6241 ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc 6301 ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct 6361 tgcccgcctg atgaatgctc atccggaatt ccgtatggca atgaaagacg gtgagctggt 6421 gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc 6481 atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga 6541 tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt 6601 tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat 6661 ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt 6721 gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag 6781 aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt aaacgcgtgg 6841 atccggctta ctaaaagcca gataacagta tgcgtatttg cgcgctgatt tttgcggtat 6901 aagaatatat actgatatgt atacccgaag tatgtcaaaa agaggtatgc tatgaagcag 6961 cgtattacag tgacagttga cagcgacagc tatcagttgc tcaaggcata tatgatgtca 7021 atatctccgg tctggtaagc acaaccatgc agaatgaagc ccgtcgtctg cgtgccgaac 7081 gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc ccggtttatt gaaatgaacg 7141 gctcttttgc tgacgagaac aggggctggt gaaatgcagt ttaaggttta cacctataaa 7201 agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga cacgcccggg 7261 cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt ctcccgtgaa 7321 ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac cgatatggcc 7381 agtgtgccgg tctccgttat cggggaagaa gtggctgatc tcagccaccg cgaaaatgac 7441 atcaaaaacg ccattaacct gatgttctgg ggaatataaa tgtcaggctc ccttatacac 7501 agccagtctg caggtcgacc atagtgactg gatatgttgt gttttacagc attatgtagt 7561 ctgtttttta tgcaaaatct aatttaatat attgatattt atatcatttt acgtttctcg 7621 ttcagctttc ttgtacaaag tggtgatggc gcgccgcggc cgcgatcaat tcggtaccga 7681 agttcctatt ccgaagttcc tattctctag aaagtatagg aacttcctcg agggtacccc 7741 aattccgccc cccccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt 7801 gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc 7861 ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag 7921 gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac 7981 aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc 8041 tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc 8101 acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca 8161 aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt 8221 gcacatgctt tacatgtgtt tagtcgaggt taaaaaacgt ctaggccccc cgaaccacgg 8281 ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg gtgagcaagg 8341 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg 8401 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc 8461 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc 8521 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct 8581 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg 8641 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg 8701 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca 8761 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga 8821 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc 8881 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc 8941 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg 9001 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaatcc gggatccgga 9061 gagctcccaa cgaagttcct attccgaagt tcctattctc tagaaagtat aggaacttcc 9121 tcgaggttgg atgcagcccg ggggatccac tagttctaga gcggccgatc agcctcgact 9181 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg 9241 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg 9301 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg 9361 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga 9421 accagctggg gctcgagatc cactagttct agcctcgagg ctagagcggc cgccgcgccg 9481 atatcgaatt cgctagaaga tgggcgggag tcttctgggc aggcttaaag gctaacctgg 9541 tgtgtgggcg ttgtcctgca ggggaattga acaggtgtaa aattggaggg acaagacttc 9601 ccacagattt tcggttttgt cgggaagttt tttaataggg gcaaataagg aaaatgggag 9661 gataggtagt catctggggt tttatgcagc aaaactacag gttattattg cttgtgatcc 9721 gcctcggagt attttccatc gaggtagatt aaagacatgc tcacccgagt tttatactct 9781 cctgcttgag atccttacta cagtatgaaa ttacagtgtc gcgagttaga ctatgtaagc 9841 agaattttaa tcatttttaa agagcccagt acttcatatc catttctccc gctccttctg 9901 cagccttatc aaaaggtatt ttagaacact cattttagcc ccattttcat ttattatact 9961 ggcttatcca acccctagac agagcattgg cattttccct ttcctgatct tagaagtctg 10021 atgactcatg aaaccagaca gattagttac atacaccaca aatcgaggct gtagctgggg 10081 cctcaacact gcagttcttt tataactcct tagtacactt tttgttgatc ctttgccttg 10141 atccttaatt ttcagtgtct atcacctctc ccgtcaggtg gtgttccaca tttgggccta 10201 ttctcagtcc agggagtttt acaacaatag atgtattgag aatccaacct aaagcttaac 10261 tttccactcc catgaatgcc tctctccttt ttctccattt ataaactgag ctattaacca 10321 ttaatggttt ccaggtggat gtctcctccc ccaatattac ctgatgtatc ttacatattg 10381 ccaggctgat attttaagac attaaaaggt atatttcatt attgagccac atggtattga 10441 ttactgctta ctaaaatttt gtcattgtac acatctgtaa aaggtggttc cttttggaat 10501 gcaaagttca ggtgtttgtt gtctttcctg acctaaggtc ttgtgagctt gtattttttc 10561 tatttaagca gtgctttctc ttggactggc ttgactcatg gcattctaca cgttattgct 10621 ggtctaaatg tgattttgcc aagcttcttc aggacctata attttgcttg acttgtagcc 10681 aaacacaagt aaaatgatta agcaacaaat gtatttgtga agcttggttt ttaggttgtt 10741 gtgttgtgtg tgcttgtgct ctataataat actatccagg ggctggagag gtggctcgga 10801 gttcaagagc acagactgct cttccagaag tcctgagttc aattcccagc aaccacatgg 10861 tggctcacaa ccatctgtaa tgggatctga tgccctcttc tggtgtgtct gaagaccaca 10921 agtgtattca cattaaataa ataaatcctc cttcttcttc tttttttttt ttttaaagag 10981 aatactgtct ccagtagaat ttactgaagt aatgaaatac tttgtgtttg ttccaatatg 11041 gtagccaata atcaaattac tctttaagca ctggaaatgt taccaaggaa ctaattttta 11101 tttgaagtgt aactgtggac agaggagcca taactgcaga cttgtgggat acagaagacc 11161 aatgcagact ttaatgtctt ttctcttaca ctaagcaata aagaaataaa aattgaactt 11221 ctagtatcct atttgtttaa actgctagct ttacttaact tttgtgcttc atctatacaa 11281 agctgaaagc taagtctgca gccattacta aacatgaaag caagtaatga taattttgga 11341 tttcaaaaat gtagggccag agtttagcca gccagtggtg gtgcttgcct ttatgccttt 11401 aatcccagca ctctggaggc agagacaggc agatctctga gtttgagccc agcctggtct 11461 acacatcaag ttctatctag gatagccagg aatacacaca gaaaccctgt tggggagggg 11521 ggctctgaga tttcataaaa ttataattga agcattccct aatgagccac tatggatgtg 11581 gctaaatccg tctacctttc tgatgagatt tgggtattat tttttctgtc tctgctgttg 11641 gttgggtctt ttgacactgt gggctttctt taaagcctcc ttcctgccat gtggtctctt 11701 gtttgctact aacttcccat ggcttaaatg gcatggcttt ttgccttcta agggcagctg 11761 ctgagatttg cagcctgatt tccagggtgg ggttgggaaa tctttcaaac actaaaattg 11821 tcctttaatt ttttttttaa aaaatgggtt atataataaa cctcataaaa tagttatgag 11881 gagtgaggtg gactaatatt aaatgagtcc ctcccctata aaagagctat taaggctttt 11941 tgtcttatac ttaacttttt ttttaaatgt ggtatcttta gaaccaaggg tcttagagtt 12001 ttagtataca gaaactgttg catcgcttaa tcagattttc tagtttcaaa tccagagaat 12061 ccaaattctt cacagccaaa gtcaaattaa gaatttctga cttttaatgt taatttgctt 12121 actgtgaata taaaaatgat agcttttcct gaggcagggt ctcactatgt atctctgcct 12181 gatctgcaac aagatatgta gactaaagtt ctgcctgctt ttgtctcctg aatactaagg 12241 ttaaaatgta gtaatacttt tggaacttgc aggtcagatt cttttatagg ggacacacta 12301 agggagcttg ggtgatagtt ggtaaaatgt gtttcaagtg atgaaaactt gaattattat 12361 caccgcaacc tactttttaa aaaaaaaagc caggcctgtt agagcatgct taagggatcc 12421 ctaggacttg ctgagcacac aagagtagtt acttggcagg ctcctggtga gagcatattt 12481 caaaaaacaa ggcagacaac caagaaacta cagttaaggt tacctgtctt taaaccatct 12541 gcatatacac agggatatta aaatattcca aataatattt cattcaagtt ttcccccatc 12601 aaattgggac atggatttct ccggtgaata ggcagagttg gaaactaaac aaatgttggt 12661 tttgtgattt gtgaaattgt tttcaagtga tagttaaagc ccatgagata cagaacaaag 12721 ctgctatttc gaggtctctt ggtttatact cagaagcact tctttgggtt tccctgcact 12781 atcctgatca tgtgctaggc ctaccttagg ctgattgttg ttcaaataaa cttaagtttc 12841 ctgtcaggtg atgtcatatg atttcatata tcaaggcaaa acatgttata tatgttaaac 12901 atttgtactt aatgtgaaag ttaggtcttt gtgggtttga tttttaattt tcaaaacctg 12961 agctaaataa gtcattttta catgtcttac atttggtgga attgtataat tgtggtttgc 13021 aggcaagact ctctgaccta gtaaccctac ctatagagca ctttgctggg tcacaagtct 13081 aggagtcaag catttcacct tgaagttgag acgttttgtt agtgtatact agtttatatg 13141 ttggaggaca tgtttatcca gaagatattc aggactattt ttgactgggc taaggaattg 13201 attctgatta gcactgttag tgagcattga gtggccttta ggcttgaatt ggagtcactt 13261 gtatatctca aataatgctg gcctttttta aaaagccctt gttctttatc accctgtttt 13321 ctacataatt tttgttcaaa gaaatacttg tttggatctc cttttgacaa caatagcatg 13381 ttttcaagcc atattttttt tccttttttt tttttttttt ggtttttcga gacagggttt 13441 ctctgtatag ccctggctgt cctggaactc actttgtaga ccaggctggc ctcgaactca 13501 gaaatccgcc tgcctctgcc tcctgagtgc cgggattaaa ggcgtgcacc accacgcctg 13561 gctaagttgg atattttgtt atataactat aaccaatact aactccactg ggtggatttt 13621 taattcagtc agtagtctta agtggtcttt attggccctt cattaaaatc tactgttcac 13681 tctaacagag gctgttggta ctagtggcac ttaagcaact tcctacggat atactagcag 13741 attaagggtc agggatagaa actagtctag cgttttgtat acctaccagc tttatactac 13801 cttgttctga tagaaatatt tcaggacatc tagcttatcg ataccgtcga cggtatcgat 13861 aagcttgata tcgaattcta ccgggtaggg gaggcgcttt tcccaaggca gtctggagca 13921 tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca 13981 cattccacat ccaccggtag gcgccgccgc cctgcagggc gatcgctacg tacagctttt 14041 gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg 14101 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 14161 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 14221 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 14281 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 14341 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 14401 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 14461 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 14521 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 14581 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 14641 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 14701 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 14761 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 14821 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 14881 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta 14941 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 15001 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 15061 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 15121 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 15181 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 15241 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 15301 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 15361 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 15421 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 15481 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 15541 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 15601 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 15661 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 15721 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 15781 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 15841 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 15901 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 15961 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 16021 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 16081 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 16141 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 16201 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt 16261 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg 16321 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg 16381 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta 16441 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg 16501 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa 16561 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct 16621 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct 16681 acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 16741 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 16801 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta 16861 atacgactca ctatagggcg aattggagct //