LOCUS Exported 12122 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Lentiviral expression of EGFR V148M. ACCESSION . VERSION . KEYWORDS pHAGE-EGFR-V148M SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12122) AUTHORS Ng PK, Li J, Jeong KJ, Shao S, Chen H, Tsang YH, Sengupta S, Wang Z, Bhavana VH, Tran R, Soewito S, Minussi DC, Moreno D, Kong K, Dogruluk T, Lu H, Gao J, Tokheim C, Zhou DC, Johnson AM, Zeng J, Ip CKM, Ju Z, Wester M, Yu S, Li Y, Vellano CP, Schultz N, Karchin R, Ding L, Lu Y, Cheung LWT, Chen K, Shaw KR, Meric-Bernstam F, Scott KL, Yi S, Sahni N, Liang H, Mills GB TITLE Systematic Functional Annotation of Somatic Mutations in Cancer. JOURNAL Cancer Cell. 2018 Mar 12;33(3):450-462.e10. doi: 10.1016/j.ccell.2018.01.021. PUBMED 29533785 REFERENCE 2 (bases 1 to 12122) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..12122 /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" promoter 2223..3401 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 2454..3392 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3349..3369 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" protein_bind 3421..3445 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 3448..7080 /codon_start=1 /product="precursor of human epidermal growth factor receptor" /label=EGFR /translation="MRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFE DHFLSLQRMFNNCEVVLGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENL QIIRGNMYYENSYALAVLSNYDANKTGLKELPMRNLQEILHGAMRFSNNPALCNVESIQ WRDIVSSDFLSNMSMDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGR CRGKSPSDCCHNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNP EGKYSFGATCVKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGI GIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVK EITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISD GDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWG PEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGP DNCIQCAHYIDGPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEG CPTNGPKIPSIATGMVGALLLLLVVALGIGLFMRRRHIVRKRTLRRLLQERELVEPLTP SGEAPNQALLRILKETEFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSP KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA" protein_bind complement(7082..7106) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" promoter 7114..7132 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(7157..7176) /label=T7 /note="T7 promoter, forward primer" promoter complement(7158..7176) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind complement(7182..7202) /gene="mutant version of attB" /label=attB4 /bound_moiety="BP Clonase(TM)" /note="core recombination site for the Gateway(R) BP reaction" CDS 7256..7297 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" misc_feature 7318..7885 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" misc_feature 7335..7887 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(7479..7496) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 7706..7725 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 7899..8498 /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(7899..7918) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 8395..8415 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 8512..9100 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(8565..8585) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(8983..8994) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 9175..9408 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" primer_bind complement(9527..9545) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 9613..9717 /gene="bla" /label=AmpR promoter CDS 9718..10578 /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(9936..9955) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 10749..11337 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 11238..11257 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 11491..11508 /label=L4440 /note="L4440 vector, forward primer" primer_bind 11600..11619 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" protein_bind 11748..11769 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 11784..11814 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 11822..11838 /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 11827..11849 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 11846..11862 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 11846..11862 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 11912..11931 /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 gactagtcgt 2221 gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg 2281 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa 2341 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg 2401 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg 2461 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa 2521 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt 2581 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct 2641 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg 2701 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct 2761 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg 2821 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc 2881 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc 2941 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac 3001 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga 3061 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc 3121 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg 3181 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga 3241 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt 3301 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga 3361 cagtggttca aagttttttt cttccatttc aggtgtcgtg aagcggccct gcagatatca 3421 acaagtttgt acaaaaaagc aggcaccatg cgaccctccg ggacggccgg ggcagcgctc 3481 ctggcgctgc tggctgcgct ctgcccggcg agtcgggctc tggaggaaaa gaaagtttgc 3541 caaggcacga gtaacaagct cacgcagttg ggcacttttg aagatcattt tctcagcctc 3601 cagaggatgt tcaataactg tgaggtggtc cttgggaatt tggaaattac ctatgtgcag 3661 aggaattatg atctttcctt cttaaagacc atccaggagg tggctggtta tgtcctcatt 3721 gccctcaaca cagtggagcg aattcctttg gaaaacctgc agatcatcag aggaaatatg 3781 tactacgaaa attcctatgc cttagcagtc ttatctaact atgatgcaaa taaaaccgga 3841 ctgaaggagc tgcccatgag aaatttacag gaaatcctgc atggcgccat gcggttcagc 3901 aacaaccctg ccctgtgcaa cgtggagagc atccagtggc gggacatagt cagcagtgac 3961 tttctcagca acatgtcgat ggacttccag aaccacctgg gcagctgcca aaagtgtgat 4021 ccaagctgtc ccaatgggag ctgctggggt gcaggagagg agaactgcca gaaactgacc 4081 aaaatcatct gtgcccagca gtgctccggg cgctgccgtg gcaagtcccc cagtgactgc 4141 tgccacaacc agtgtgctgc aggctgcaca ggcccccggg agagcgactg cctggtctgc 4201 cgcaaattcc gagacgaagc cacgtgcaag gacacctgcc ccccactcat gctctacaac 4261 cccaccacgt accagatgga tgtgaacccc gagggcaaat acagctttgg tgccacctgc 4321 gtgaagaagt gtccccgtaa ttatgtggtg acagatcacg gctcgtgcgt ccgagcctgt 4381 ggggccgaca gctatgagat ggaggaagac ggcgtccgca agtgtaagaa gtgcgaaggg 4441 ccttgccgca aagtgtgtaa cggaataggt attggtgaat ttaaagactc actctccata 4501 aatgctacga atattaaaca cttcaaaaac tgcacctcca tcagtggcga tctccacatc 4561 ctgccggtgg catttagggg tgactccttc acacatactc ctcctctgga tccacaggaa 4621 ctggatattc tgaaaaccgt aaaggaaatc acagggtttt tgctgattca ggcttggcct 4681 gaaaacagga cggacctcca tgcctttgag aacctagaaa tcatacgcgg caggaccaag 4741 caacatggtc agttttctct tgcagtcgtc agcctgaaca taacatcctt gggattacgc 4801 tccctcaagg agataagtga tggagatgtg ataatttcag gaaacaaaaa tttgtgctat 4861 gcaaatacaa taaactggaa aaaactgttt gggacctccg gtcagaaaac caaaattata 4921 agcaacagag gtgaaaacag ctgcaaggcc acaggccagg tctgccatgc cttgtgctcc 4981 cccgagggct gctggggccc ggagcccagg gactgcgtct cttgccggaa tgtcagccga 5041 ggcagggaat gcgtggacaa gtgcaacctt ctggagggtg agccaaggga gtttgtggag 5101 aactctgagt gcatacagtg ccacccagag tgcctgcctc aggccatgaa catcacctgc 5161 acaggacggg gaccagacaa ctgtatccag tgtgcccact acattgacgg cccccactgc 5221 gtcaagacct gcccggcagg agtcatggga gaaaacaaca ccctggtctg gaagtacgca 5281 gacgccggcc atgtgtgcca cctgtgccat ccaaactgca cctacggatg cactgggcca 5341 ggtcttgaag gctgtccaac gaatgggcct aagatcccgt ccatcgccac tgggatggtg 5401 ggggccctcc tcttgctgct ggtggtggcc ctggggatcg gcctcttcat gcgaaggcgc 5461 cacatcgttc ggaagcgcac gctgcggagg ctgctgcagg agagggagct tgtggagcct 5521 cttacaccca gtggagaagc tcccaaccaa gctctcttga ggatcttgaa ggaaactgaa 5581 ttcaaaaaga tcaaagtgct gggctccggt gcgttcggca cggtgtataa gggactctgg 5641 atcccagaag gtgagaaagt taaaattccc gtcgctatca aggaattaag agaagcaaca 5701 tctccgaaag ccaacaagga aatcctcgat gaagcctacg tgatggccag cgtggacaac 5761 ccccacgtgt gccgcctgct gggcatctgc ctcacctcca ccgtgcagct catcacgcag 5821 ctcatgccct tcggctgcct cctggactat gtccgggaac acaaagacaa tattggctcc 5881 cagtacctgc tcaactggtg tgtgcagatc gcaaagggca tgaactactt ggaggaccgt 5941 cgcttggtgc accgcgacct ggcagccagg aacgtactgg tgaaaacacc gcagcatgtc 6001 aagatcacag attttgggct ggccaaactg ctgggtgcgg aagagaaaga ataccatgca 6061 gaaggaggca aagtgcctat caagtggatg gcattggaat caattttaca cagaatctat 6121 acccaccaga gtgatgtctg gagctacggg gtgaccgttt gggagttgat gacctttgga 6181 tccaagccat atgacggaat ccctgccagc gagatctcct ccatcctgga gaaaggagaa 6241 cgcctccctc agccacccat atgtaccatc gatgtctaca tgatcatggt caagtgctgg 6301 atgatagacg cagatagtcg cccaaagttc cgtgagttga tcatcgaatt ctccaaaatg 6361 gcccgagacc cccagcgcta ccttgtcatt cagggggatg aaagaatgca tttgccaagt 6421 cctacagact ccaacttcta ccgtgccctg atggatgaag aagacatgga cgacgtggtg 6481 gatgccgacg agtacctcat cccacagcag ggcttcttca gcagcccctc cacgtcacgg 6541 actcccctcc tgagctctct gagtgcaacc agcaacaatt ccaccgtggc ttgcattgat 6601 agaaatgggc tgcaaagctg tcccatcaag gaagacagct tcttgcagcg atacagctca 6661 gaccccacag gcgccttgac tgaggacagc atagacgaca ccttcctccc agtgcctgaa 6721 tacataaacc agtccgttcc caaaaggccc gctggctctg tgcagaatcc tgtctatcac 6781 aatcagcctc tgaaccccgc gcccagcaga gacccacact accaggaccc ccacagcact 6841 gcagtgggca accccgagta tctcaacact gtccagccca cctgtgtcaa cagcacattc 6901 gacagccctg cccactgggc ccagaaaggc agccaccaaa ttagcctgga caaccctgac 6961 taccagcagg acttctttcc caaggaagcc aagccaaatg gcatctttaa gggctccaca 7021 gctgaaaatg cagaatacct aagggtcgcg ccacaaagca gtgaatttat tggagcatag 7081 aacccagctt tcttgtacaa agtggtgata tccaattaac cctcactaaa gggaatgttt 7141 gtaaatgtat agatagccct atagtgagtc gtattaccac ccaacttttc tatacaaagt 7201 ggttgatatc cagcacagtg gcggccgctc gagtctagag ggcccgcggt tcgaaggtaa 7261 gcctatccct aaccctctcc tcggtctcga ttctacgcgt accggttagt aatgagatcc 7321 ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt 7381 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg 7441 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg 7501 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc 7561 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca 7621 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag 7681 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa 7741 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt 7801 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt 7861 tttcctttga aaaacacgat gataatatgg ccacacatat gaccgagtac aagcccacgg 7921 tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg 7981 ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg 8041 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg 8101 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt 8161 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac 8221 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg 8281 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag 8341 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc 8401 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac 8461 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctaaat cgatagatcc taatcaacct 8521 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 8581 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 8641 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 8701 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 8761 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 8821 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 8881 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt 8941 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 9001 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 9061 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctgagatcct ttaagaccaa 9121 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag 9181 ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt 9241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc 9301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta 9361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat 9421 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagaggc 9481 ccgggttaat taaggaaagg gctagatcat tcttgaagac gaaagggcct cgtgatacgc 9541 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt 9601 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat 9661 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg 9721 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt 9781 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga 9841 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa 9901 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt 9961 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt 10021 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc 10081 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga 10141 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat 10201 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct 10261 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc 10321 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg 10381 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc 10441 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg 10501 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca 10561 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta 10621 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 10681 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 10741 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 10801 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 10861 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 10921 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 10981 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 11041 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 11101 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 11161 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 11221 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 11281 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 11341 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc 11401 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat 11461 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag 11521 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagcaagctc 11581 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat 11641 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctccccgtgg 11701 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag 11761 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga 11821 attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac gaatttcaca 11881 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 11941 tatcatgtct ggatcaactg gataactcaa gctaaccaaa atcatcccaa acttcccacc 12001 ccatacccta ttaccactgc caattacctg tggtttcatt tactctaaac ctgtgattcc 12061 tctgaattat tttcatttta aagaaattgt atttgttaaa tatgtactac aaacttagta 12121 gt //