LOCUS Exported 11900 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAVS1 donor vector for inducible gene expression. ACCESSION . VERSION . KEYWORDS AAVS1-TRE3G-EGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11900) AUTHORS Qian K, Huang CL, Chen H, Blackbourn LW 4th, Chen Y, Cao J, Yao L, Sauvey C, Du Z, Zhang SC TITLE A simple and efficient system for regulating gene expression in human pluripotent stem cells and derivatives. JOURNAL Stem Cells. 2014 Feb 4. doi: 10.1002/stem.1653. PUBMED 24497442 REFERENCE 2 (bases 1 to 11900) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11900 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 37..840 /label=HA-L /note="left homology arm from the adeno-associated virus integration site (AAVS1) within intron 1 of the human PPP1R12C gene" misc_feature 847..872 /label=SA /note="splice acceptor site" CDS 896..949 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 959..1558 /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(959..978) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 1455..1475 /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind complement(1590..1607) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1596..1820 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" polyA_signal 1858..1979 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1869..1888) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1923..1942 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS complement(2404..2424) /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" intron 2554..2619 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS complement(2780..3499) /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 complement(2783..3499) /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2825..2846) /label=EGFP-C /note="EGFP, forward primer" primer_bind 3174..3193 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3433..3454 /label=EGFP-N /note="EGFP, reverse primer" promoter complement(3506..3884) /label=TRE3G promoter /note="3rd-generation Tet-responsive promoter that can be activated by binding of Tet-On(R) 3G" primer_bind complement(3573..3597) /label=LNCX /note="Human CMV promoter, forward primer" protein_bind 3642..3660 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 3678..3696 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 3714..3732 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 3750..3768 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 3786..3804 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 3822..3840 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 3858..3876 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" enhancer 3968..4271 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4273..4548 /label=chicken beta-actin promoter intron 4549..5565 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 5489..5511 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 5573..5592 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 5630..6376 /codon_start=1 /product="modified rtTA protein that binds tightly to promoters containing the tet operator in the presence of doxycycline" /label=Tet-On(R) 3G /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA DALDDFDLDMLPADALDDFDLDMLPG" primer_bind complement(6467..6486) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 6532..6587 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(6586..6605) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(6948..6964) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 6972..6988 /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(6996..7026) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7041..7062 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." misc_feature 7136..7972 /label=HA-R /note="right homology arm from the adeno-associated virus integration site (AAVS1) within intron 1 of the human PPP1R12C gene" primer_bind complement(8006..8025) /label=T7 /note="T7 promoter, forward primer" promoter complement(8007..8025) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(8032..8049) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(8032..8048) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8041..8063) /label=M13/pUC Forward /note="In lacZ gene" CDS 8186..8488 /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="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind 8367..8386 /label=ccdB-fwd /note="ccdB gene, forward primer" primer_bind complement(8891..8910) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 9505..9524 /label=Neo-F /note="Neomycin resistance gene, forward primer" CDS complement(9885..10745) /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 10508..10527 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 10869..11457 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 11358..11377 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 11611..11628 /label=L4440 /note="L4440 vector, forward primer" protein_bind 11745..11766 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 11781..11811 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 11819..11835 /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)." primer_bind 11843..11859 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" promoter 11880..11898 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" ORIGIN 1 ctagctagtc ctgcaggttt aaacgaattc gccctttgct ttctctgacc agcattctct 61 cccctgggcc tgtgccgctt tctgtctgca gcttgtggcc tgggtcacct ctacggctgg 121 cccagatcct tccctgccgc ctccttcagg ttccgtcttc ctccactccc tcttcccctt 181 gctctctgct gtgttgctgc ccaaggatgc tctttccgga gcacttcctt ctcggcgctg 241 caccacgtga tgtcctctga gcggatcctc cccgtgtctg ggtcctctcc gggcatctct 301 cctccctcac ccaaccccat gccgtcttca ctcgctgggt tcccttttcc ttctccttct 361 ggggcctgtg ccatctctcg tttcttagga tggccttctc cgacggatgt ctcccttgcg 421 tcccgcctcc ccttcttgta ggcctgcatc atcaccgttt ttctggacaa ccccaaagta 481 ccccgtctcc ctggctttag ccacctctcc atcctcttgc tttctttgcc tggacacccc 541 gttctcctgt ggattcgggt cacctctcac tcctttcatt tgggcagctc ccctaccccc 601 cttacctctc tagtctgtgc tagctcttcc agccccctgt catggcatct tccaggggtc 661 cgagagctca gctagtcttc ttcctccaac ccgggcccct atgtccactt caggacagca 721 tgtttgctgc ctccagggat cctgtgtccc cgagctggga ccaccttata ttcccagggc 781 cggttaatgt ggctctggtt ctgggtactt ttatctgtcc cctccacccc acagtggggc 841 aagcttctga cctcttctct tcctcccaca gggcctcgag agatctggca gcggagaggg 901 cagaggaagt cttctaacat gcggtgacgt ggaggagaat cccggcccta ggctcgagat 961 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg 1021 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg 1081 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat 1141 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag 1201 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc 1261 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc 1321 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag 1381 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga 1441 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga 1501 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgatc 1561 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc 1621 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct 1681 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg 1741 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg 1801 ggatgcggtg ggctctatgg gtctcgacat tgattattga ctaggccgca gacatgataa 1861 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt 1921 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta 1981 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt 2041 aaagcaagta aaacctctac aaatgtggta tggctgatta tgatctctag tcaaggcact 2101 atacatcaaa tattccttat taaccccttt acaaattaaa aagctaaagg tacacaattt 2161 ttgagcatag ttattaatag cagacactct atgcctgtgt ggagtaagaa aaaacagtat 2221 gttatgatta taactgttat gcctacttat aaaggttaca gaatattttt ccataatttt 2281 cttgtatagc agtgcagctt tttcctttgt ggtgtaaata gcaaagcaag caagagttct 2341 attactaaac acagcatgac tcaaaaaact tagcaattct gaaggaaagt ccttggggtc 2401 ttctaccttt ctcttctttt ttggaggagt agaatgttga gagtcagcag tagcctcatc 2461 atcactagat ggcatttctt ctgagcaaaa caggttttcc tcattaaagg cattccacca 2521 ctgctcccat tcatcagttc cataggttgg aatctaaaat acacaaacaa ttagaatcag 2581 tagtttaaca cattatacac ttaaaaattt tatatttacc ttagagcttt aaatctctgt 2641 aggtagtttg tccaattatg tcacaccaca gaagtaaggt tccttcacaa agatcctcta 2701 gagtattaca atagctaaga atttcgtcat cgctgaatac agttacattg gatccctgca 2761 ggctagccat atgacgcgtt tacttgtaca gctcgtccat gccgagagtg atcccggcgg 2821 cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcaggg 2881 cggactgggt gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg 2941 tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct 3001 tgaagttcac cttgatgccg ttcttctgct tgtcggccat gatatagacg ttgtggctgt 3061 tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct 3121 tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt 3181 agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg 3241 acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt 3301 aggtcagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga 3361 acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact 3421 tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct 3481 cctcgccctt gctcaccatg tcgactttac gagggtagga agtggtacgg aaagttggta 3541 taagacaaaa gtgttgtgga attgctccag gcgatctgac ggttcactaa acgagctctg 3601 cttttatagg cgcccaccgt acacgcctaa agcttatacg ttctctatca ctgataggga 3661 gtaaactgga tatacgttct ctatcactga tagggagtaa actgtagata cgttctctat 3721 cactgatagg gagtaaactg gtcatacgtt ctctatcact gatagggagt aaactcctta 3781 tacgttctct atcactgata gggagtaaag tctgcatacg ttctctatca ctgataggga 3841 gtaaactctt catacgttct ctatcactga tagggagtaa actcgaggaa ttcttgaaga 3901 cgaaaactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg 3961 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc 4021 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt 4081 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc 4141 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg 4201 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg 4261 ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct 4321 ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg 4381 gggggggggg ggsgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg 4441 cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg 4501 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg 4561 cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga 4621 ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat 4681 tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg 4741 ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc 4801 gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc 4861 gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg 4921 cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg 4981 tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga 5041 gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc 5101 gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg 5161 gccggggagg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc 5221 gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct 5281 ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg 5341 cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg 5401 tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc 5461 tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct 5521 agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg 5581 ctggttattg tgctgtctca tcattttggc aaagaattcg tcgagcacca tgtctagact 5641 ggacaagagc aaagtcataa actctgctct ggaattactc aatggagtcg gtatcgaagg 5701 cctgacgaca aggaaactcg ctcaaaagct gggagttgag cagcctaccc tgtactggca 5761 cgtgaagaac aagcgggccc tgctcgatgc cctgccaatc gagatgctgg acaggcatca 5821 tacccactcc tgccccctgg aaggcgagtc atggcaagac tttctgcgga acaacgccaa 5881 gtcataccgc tgtgctctcc tctcacatcg cgacggggct aaagtgcatc tcggcacccg 5941 cccaacagag aaacagtacg aaaccctgga aaatcagctc gcgttcctgt gtcagcaagg 6001 cttctccctg gagaacgcac tgtacgctct gtccgccgtg ggccacttta cactgggctg 6061 cgtattggag gaacaggagc atcaagtagc aaaagaggaa agagagacac ctaccaccga 6121 ttctatgccc ccacttctga aacaagcaat tgagctgttc gaccggcagg gagccgaacc 6181 tgccttcctt ttcggcctgg aactaatcat atgtggcctg gagaaacagc taaagtgcga 6241 aagcggcggg ccgaccgacg cccttgacga ttttgactta gacatgctcc cagccgatgc 6301 ccttgacgac tttgaccttg atatgctgcc tgctgacgct cttgacgatt ttgaccttga 6361 catgctcccc gggtaaggcg cgtgaattca ctcctcaggt gcaggctgcc tatcagaagg 6421 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg 6481 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga 6541 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat 6601 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat 6661 atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga 6721 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt 6781 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg 6841 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc 6901 tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt 6961 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag 7021 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg 7081 cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcg acagtactaa gctttactag 7141 ggacaggatt ggtgacagaa aagccccatc cttaggcctc ctccttccta gtctcctgat 7201 attgggtcta acccccacct cctgttaggc agattcctta tctggtgaca cacccccatt 7261 tcctggagcc atctctctcc ttgccagaac ctctaaggtt tgcttacgat ggagccagag 7321 aggatcctgg gagggagagc ttggcagggg gtgggaggga agggggggat gcgtgacctg 7381 cccggttctc agtggccacc ctgcgctacc ctctcccaga acctgagctg ctctgacgcg 7441 gctgtctggt gcgtttcact gatcctggtg ctgcagcttc cttacacttc ccaagaggag 7501 aagcagtttg gaaaaacaaa atcagaataa gttggtcctg agttctaact ttggctcttc 7561 acctttctag tccccaattt atattgttcc tccgtgcgtc agttttacct gtgagataag 7621 gccagtagcc agccccgtcc tggcagggct gtggtgagga ggggggtgtc cgtgtggaaa 7681 actccctttg tgagaatggt gcgtcctagg tgttcaccag gtcgtggccg cctctactcc 7741 ctttctcttt ctccatcctt ctttccttaa agagtcccca gtgctatctg ggacatattc 7801 ctccgcccag agcagggtcc cgcttcccta aggccctgct ctgggcttct gggtttgagt 7861 ccttggcaag cccaggagag gcgctcaggc ttccctgtcc cccttcctcg tccaccatct 7921 catgcccctg gctctcctgc cccttcccta caggggttcc tggctctgct ctaagggcga 7981 attcgcggcc gctaaattca attcgcccta tagtgagtcg tattacaatt cactggccgt 8041 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc 8101 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca 8161 acagttgcgc agcctatacg tacggcagtt taaggtttac acctataaaa gagagagccg 8221 ttatcgtctg tttgtggatg tacagagtga tattattgac acgccggggc gacggatggt 8281 gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac tttacccggt 8341 ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca gtgtgccggt 8401 ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc 8461 cattaacctg atgttctggg gaatataaat gtcaggcatg agattatcaa aaaggatctt 8521 cacctagatc cttttcacgt agaaagccag tccgcagaaa cggtgctgac cccggatgaa 8581 tgtcagctac tgggctatct ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc 8641 ttgcagtggg cttacatggc gatagctaga ctgggcggtt ttatggacag caagcgaacc 8701 ggaattgcca gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat 8761 ggctttcttg ccgccaagga tctgatggcg caggggatca agctctgatc aagagacagg 8821 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg 8881 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc 8941 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg 9001 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt 9061 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg 9121 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat 9181 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca 9241 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca 9301 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa 9361 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat gcatggcgat gcctgcttgc 9421 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg 9481 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg 9541 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca 9601 tcgccttcta tcgccttctt gacgagttct tctgaattat taacgcttac aatttcctga 9661 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatcagg tggcactttt 9721 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat 9781 ccgctcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 9841 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 9901 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 9961 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 10021 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 10081 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 10141 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 10201 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 10261 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 10321 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 10381 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgaatac 10441 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 10501 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 10561 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 10621 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 10681 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 10741 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgacc 10801 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 10861 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 10921 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 10981 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 11041 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 11101 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 11161 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 11221 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 11281 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 11341 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 11401 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 11461 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc 11521 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat 11581 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag 11641 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac 11701 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc 11761 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt 11821 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctcaga 11881 attaaccctc actaaaggga //