LOCUS Exported 12975 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Lentiviral expression of human MEFV M694V after doxycycline administration. ACCESSION . VERSION . KEYWORDS p21-M694V-MEFV SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12975) AUTHORS Magnotti F, Lefeuvre L, Benezech S, Malsot T, Waeckel L, Martin A, Kerever S, Chirita D, Desjonqueres M, Duquesne A, Gerfaud-Valentin M, Laurent A, Seve P, Popoff MR, Walzer T, Belot A, Jamilloux Y, Henry T TITLE Pyrin dephosphorylation is sufficient to trigger inflammasome activation in familial Mediterranean fever patients. JOURNAL EMBO Mol Med. 2019 Oct 7:e10547. doi: 10.15252/emmm.201910547. PUBMED 31589380 REFERENCE 2 (bases 1 to 12975) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..12975 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 1..634 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 681..806 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1303..1536 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1721..1765 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 2058..2175 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" protein_bind 2230..2500 /label=tetracycline response element /bound_moiety="tetracycline repressor TetR" /note="contains seven copies of the tetracycline operator tetO" promoter 2533..2571 /label=minimal CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2568..2592 /label=LNCX /note="Human CMV promoter, forward primer" protein_bind 2701..2725 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2768..2833 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" protein_bind complement(5203..5227) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 5280..5306 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" promoter 5526..6704 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 5757..6695 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 6652..6672 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" protein_bind complement(6774..6807) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS 6814..7560 /codon_start=1 /product="improved tetracycline-controlled transactivator" /label=rtTA-Advanced /note="In the Tet-On(R) system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline." /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA DALDDFDLDMLPADALDDFDLDMLPG" misc_feature 7620..8187 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(7781..7798) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 8008..8027 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 8203..8922 /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 8203..8919 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(8248..8269) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(8509..8528) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 8856..8877 /label=EGFP-C /note="EGFP, forward primer" primer_bind 9058..9077 /label=Neo-F /note="Neomycin resistance gene, forward primer" misc_feature 9365..9953 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(9418..9438) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(9836..9847) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 10028..10261 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" primer_bind complement(10380..10398) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 10466..10570 /gene="bla" /label=AmpR promoter CDS 10571..11431 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(10789..10808) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 11602..12190 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 12091..12110 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 12344..12361 /label=L4440 /note="L4440 vector, forward primer" primer_bind 12453..12472 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" protein_bind 12601..12622 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 12637..12667 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 12675..12691 /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 12680..12702 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 12699..12715 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 12699..12715 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 12765..12784 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg 2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa 2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca 2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactaggatc 2221 ctttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca 2281 gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa 2341 agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc 2401 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa 2461 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagctcgg 2521 tacccgggtc gaggtaggcg tgtacggtgg gaggcctata taagcagagc tcgtttagtg 2581 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg 2641 gaccgatcca gcctccgcgg ccccgaacta gtccagtgtg gtggaattct gcagatatca 2701 acaagtttgt acaaaaaagc aggctttaaa ggaaccaatt cagtcgactg gatccggtac 2761 caccatggac tacaaagacc atgacggtga ttataaagat catgacatcg attacaagga 2821 tgacgatgac aagcttgcgg ccgcggctaa gacccctagt gaccatctgc tgtccaccct 2881 ggaggagctg gtgccctatg acttcgagaa gttcaagttc aagctgcaga acaccagtgt 2941 gcagaaggag cactccagga tcccccggag ccagatccag agagccaggc cggtgaagat 3001 ggccactctg ctggtcacct actatgggga agagtacgcc gtgcagctca ccctgcaggt 3061 cctgcgggcc atcaaccagc gcctgctggc cgaggagctc cacagggcag ccattcagga 3121 atattccaca caagaaaacg gcacagatga ttccgcagcg tccagctccc tgggggagaa 3181 caagcccagg agcctgaaga ctccagacca ccccgagggg aacgagggga acggccctcg 3241 gccgtacggg ggcggagctg ccagcctgcg gtgcagccag cccgaggccg ggagggggct 3301 gtcgaggaag cccctgagca aacgcagaga gaaggcctcg gagggcctgg acgcgcaggg 3361 caagcctcgg acccggagcc cggccctgcc gggcgggaga agccccggcc cctgcagggc 3421 gctagagggg ggccaggccg aggtccggct gcgcagaaac gccagctccg cggggaggct 3481 gcaggggctg gcggggggcg ccccggggca gaaggagtgc aggcccttcg aagtgtacct 3541 gccctcggga aagatgcgac ctagaagcct tgaggtcacc atttctacag gggagaaggc 3601 gcccgcaaat ccagaaattc tcctgactct agaggaaaag acagctgcga atctggactc 3661 ggcaacagaa ccccgggcaa ggcccactcc ggatggaggg gcatctgcgg acctgaagga 3721 aggccctgga aatccagaac attcggtcac cggaaggcca ccagacacgg ctgcgagtcc 3781 ccgctgccac gcccaggaag gagacccagt tgacggtacc tgtgtgcgtg attcctgcag 3841 cttccccgag gcagtttctg ggcaccccca ggcctcaggc agccgctcac ctggctgccc 3901 ccggtgccag gactcccatg aaaggaagag cccgggaagc ctaagccccc agcccctgcc 3961 acagtgtaag cgccacctga agcaggtcca gctgctcttc tgtgaggatc acgatgagcc 4021 catctgcctc atctgcagtc tgagtcagga gcaccaaggc caccgggtgc gccccattga 4081 ggaggtcgcc ctggaacaca agaagaaaat tcagaagcag ctggagcatc tgaagaagct 4141 gagaaaatca ggggaggagc agcgatccta tggggaggag aaggcagtga gctttctgaa 4201 acaaactgaa gcgctgaagc agcgggtgca gaggaagctg gagcaggtgt actacttcct 4261 ggagcagcaa gagcatttct ttgtggcctc actggaggac gtgggccaga tggttgggca 4321 gatcaggaag gcatatgaca cccgcgtatc ccaggacatc gccctgctcg atgcgctgat 4381 tggggaactg gaggccaagg agtgccagtc agaatgggaa cttctgcagg acattggaga 4441 catcttgcac agggctaaga cagtgcctgt ccctgaaaag tggaccactc ctcaagagat 4501 aaaacaaaag atccaactcc tccaccagaa gtcagagttt gtggagaaga gcacaaagta 4561 cttctcagaa accctgcgtt cagaaatgga aatgttcaat gttccagagc tgattggcgc 4621 tcaggcacat gctgttaatg tgattctgga tgcagaaacc gcttacccca acctcatctt 4681 ctctgatgat ctgaagagtg ttagacttgg aaacaagtgg gagaggctgc ctgatggccc 4741 gcaaagattt gacagctgta tcattgttct gggctctccg agtttcctct ctggccgccg 4801 ttactgggag gtggaggttg gagacaagac agcatggatc ctgggagcct gcaagacatc 4861 cataagcagg aaagggaaca tgactctgtc gccagagaat ggctactggg tggtgataat 4921 ggtgaaggaa aatgagtacc aggcgtccag cgttcccccg acccgcctgc taataaagga 4981 gcctcccaag cgtgtgggca tcttcgtgga ctacagagtt ggaagcatct ccttttacaa 5041 tgtgacagcc agatcccaca tctatacatt cgccagctgc tctttctctg ggccccttca 5101 acctatcttc agccctggga cacgtgatgg agggaagaac acagctcctc tgactatctg 5161 tccagtgggt ggtcaggggc ctgactgact cgagatatct agacccagct ttcttgtaca 5221 aagtggttga tatccagcac agtggcggcc gctcgagtct agagggcccg cggttcgaat 5281 acccatacga cgtcccagac tacgcttagt aatgattaat taaactagaa attctaccgg 5341 gtaggggagg cgcttttccc aaggcagtct ggagtaaccc acccaagatc tggcctccgc 5401 gccgggtttt ggcgcctccc gcgggcgccc ccctcctcac ggcgagccgc gtaagatctg 5461 gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcctcacgg cgagcctagt 5521 cgtgaggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt 5581 tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg 5641 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa 5701 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtaa 5761 gtgccgtgtg tggttcccgc gggcctggcc tctttacggg ttatggccct tgcgtgcctt 5821 gaattacttc cacctggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg 5881 ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg 5941 cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg 6001 ctgctttcga taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt 6061 tctggcaaga tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg 6121 gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc 6181 tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg cctgctctgg 6241 tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg 6301 caccagttgc gtgagcggaa agatggccgc ttcccggccc tgctgcaggg agctcaaaat 6361 ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct 6421 ttccgtcctc agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc 6481 tcgattagtt ctcgagcttt tggagtacgt cgtctttagg ttggggggag gggttttatg 6541 cgatggagtt tccccacact gagtgggtgg agactgaagt taggccagct tggcacttga 6601 tgtaattctc cttggaattt gccctttttg agtttggatc ttggttcatt ctcaagcctc 6661 agacagtggt tcaaagtttt tttcttccat ttcaggtgtc gtgaagcggc cgccatgcta 6721 gtaaattgtc cgctaaattc tggccgtttt tggctttttt gttagacgga tccataactt 6781 cgtataatgt atgctatacg aagttatgcc accatgtcta gactggacaa gagcaaagtc 6841 ataaacggag ctctggaatt actcaatggt gtcggtatcg aaggcctgac gacaaggaaa 6901 ctcgctcaaa agctgggagt tgagcagcct accctgtact ggcacgtgaa gaacaagcgg 6961 gccctgctcg atgccctgcc aatcgagatg ctggacaggc atcataccca cttctgcccc 7021 ctggaaggcg agtcatggca agactttctg cggaacaacg ccaagtcata ccgctgtgct 7081 ctcctctcac atcgcgacgg ggctaaagtg catctcggca cccgcccaac agagaaacag 7141 tacgaaaccc tggaaaatca gctcgcgttc ctgtgtcagc aaggcttctc cctggagaac 7201 gcactgtacg ctctgtccgc cgtgggccac tttacactgg gctgcgtatt ggaggaacag 7261 gagcatcaag tagcaaaaga ggaaagagag acacctacca ccgattctat gcccccactt 7321 ctgagacaag caattgagct gttcgaccgg cagggagccg aacctgcctt ccttttcggc 7381 ctggaactaa tcatatgtgg cctggagaaa cagctaaagt gcgaaagcgg cgggccgacc 7441 gacgcccttg acgattttga cttagacatg ctcccagccg atgcccttga cgactttgac 7501 cttgatatgc tgcctgctga cgctcttgac gattttgacc ttgacatgct ccccgggtaa 7561 ctaagtaagg atccgcggcc gcactagagg aattccgccc ctctcccccc cccccnntct 7621 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt 7681 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct 7741 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa 7801 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg 7861 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc 7921 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg 7981 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg 8041 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc 8101 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg 8161 gttttccttt gaaaaacacg atgataagct taccggtcca ccatggtgag caagggcgag 8221 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac 8281 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag 8341 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc 8401 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag 8461 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac 8521 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg 8581 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac 8641 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc 8701 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac 8761 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc 8821 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc 8881 gccgccggga tcactctcgg catggacgag ctgtacaagt aagcatgccc gacggcgagg 8941 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 9001 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 9061 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 9121 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 9181 tcttctgatg tacaagtaaa gcggccgcga ctctagatca taatcagcca taccacattt 9241 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa 9301 atgaatgcaa ttgttgttgt ttagtccctc ccaattcgat atcaagctta tcgcgataga 9361 tcctaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 9421 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 9481 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 9541 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 9601 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 9661 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 9721 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct 9781 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 9841 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 9901 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctgagat 9961 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag 10021 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact 10081 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca 10141 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg 10201 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc 10261 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca 10321 gagagtgaga ggcccgggtt aattaaggaa agggctagat cattcttgaa gacgaaaggg 10381 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc 10441 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca 10501 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa 10561 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt 10621 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca 10681 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag 10741 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc 10801 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca 10861 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt 10921 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct 10981 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt 11041 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga 11101 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact 11161 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc 11221 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga 11281 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt 11341 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga 11401 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact 11461 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga 11521 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt 11581 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca 11641 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct 11701 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta 11761 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct 11821 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc 11881 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca 11941 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga 12001 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg 12061 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt 12121 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag 12181 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt 12241 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt 12301 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga 12361 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta 12421 atgcagcaag ctcatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 12481 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa 12541 aagctccccg tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat 12601 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg 12661 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat 12721 tacgaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact 12781 catcaatgta tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc 12841 caaacttccc accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta 12901 aacctgtgat tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac 12961 tacaaactta gtagt //