LOCUS Exported 15002 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentiviral vector for doxycycline-inducible expression of membrane targeted-CSK in mammalian cells. ACCESSION . VERSION . KEYWORDS Tet-myr-CSK-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 15002) AUTHORS Kim NG, Gumbiner BM TITLE Adhesion to fibronectin regulates Hippo signaling via the FAK-Src-PI3K pathway. JOURNAL J Cell Biol. 2015 Aug 3;210(3):503-15. doi: 10.1083/jcb.201501025. Epub 2015 Jul 27. PUBMED 26216901 REFERENCE 2 (bases 1 to 15002) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..15002 /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 2063..2180 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" CDS complement(2245..2619) /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" regulatory 2616..2625 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" promoter complement(2629..2647) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter complement(2639..2686) /label=EM7 promoter /note="synthetic bacterial promoter " protein_bind 2722..2740 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2757..2775 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2793..2811 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2829..2847 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2864..2882 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2900..2918 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" promoter 2930..2968 /label=minimal CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2965..2989 /label=LNCX /note="Human CMV promoter, forward primer" protein_bind 3009..3033 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 3033..3074 /codon_start=1 /product="N-myristoylation signal from Src kinase (Pellman et al., 1985; Kaplan et al., 1988)" /label=myr /translation="MGSSKSKPKDPSQR" regulatory 4437..4446 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4443..5162 /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 4443..5159 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(4488..4509) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4749..4768) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5096..5117 /label=EGFP-C /note="EGFP, forward primer" protein_bind complement(5162..5186) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 5194..5217 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 5221..5244 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" ncRNA 5309..5430 /label=5' miR-30a /note="sequence upstream of the 71-nt precursor of the human miR-30a microRNA (Zeng et al., 2002)" /ncRNA_class="miRNA" ncRNA 5458..5472 /label=miR-30a loop /note="loop from the the 71-nt precursor of the human miR-30a microRNA (Zeng et al., 2002)" /ncRNA_class="miRNA" ncRNA 5500..5626 /label=3' miR-30a /note="sequence downstream of the 71-nt precursor of the human miR-30a microRNA (Zeng et al., 2002)" /ncRNA_class="miRNA" primer_bind complement(5720..5739) /label=T7 /note="T7 promoter, forward primer" promoter complement(5721..5739) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter 5752..6963 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 6800..6819 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" protein_bind 6976..7009 /label=loxP511 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GTATACAT). loxP511 sites are compatible with each other, but incompatible with wild-type loxP sites." CDS 7016..7723 /codon_start=1 /product="reverse tetracycline transactivator 3" /label=rtTA3 /note="activates expression from Tet-responsive promoters in the presence of doxycycline" /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA DALDDFDLDMLPG" protein_bind 7724..7757 /label=loxP511 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GTATACAT). loxP511 sites are compatible with each other, but incompatible with wild-type loxP sites." misc_feature 7780..8352 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(7947..7964) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 8174..8193 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 8355..8957 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MATEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGSRMAELSGSRL AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind 8854..8874 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 8973..9561 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(9026..9046) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(9444..9455) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 9768..10001 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" primer_bind complement(10102..10119) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 10108..10332 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 10378..10806 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(10465..10484) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 10675..10696 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(10815..10835) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 10820..11149 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 11000..11135 /label=SV40 ori /note="SV40 origin of replication" primer_bind 11062..11081 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 12357..12478 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(12527..12543) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(12527..12543) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(12540..12562) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 12551..12567 /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(12575..12605) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 12620..12641 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(12758..12775) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(12929..13517) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(13009..13028) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(13688..14548) /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 14311..14330 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(14549..14653) /gene="bla" /label=AmpR promoter polyA_signal 14701..14835 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" 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 atcggcactg cgtgcgccaa ttctgcagac 2041 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag 2101 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa 2161 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttag 2221 taccgggccc gctctagtcc ggaatcagtc ctgctcctcg gccacgaagt gcacgcagtt 2281 gccggccggg tcgcgcaggg cgaactcccg cccccacggc tgctcgccga tctcggtcat 2341 ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc tccgaccact cggcgtacag 2401 ctcgtccagg ccgcgcaccc acacccaggc cagggtgttg tccggcacca cctggtcctg 2461 gaccgcgctg atgaacaggg tcacgtcgtc ccggaccaca ccggcgaagt cgtcctccac 2521 gaagtcccgg gagaacccga gccggtcggt ccagaactcg accgctccgg cgacgtcgcg 2581 cgcggtgagc accggaacgg cactggtcaa cttggccatg gtggccctcc tatagtgagt 2641 cgtattatac tatgccgata tactatgccg atgattaatt gtcaacacgt gctgcaggtc 2701 cgaggttcta gacgagttta ctccctatca gtgatagaga acgatgtcga gtttactccc 2761 tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc 2821 gagtttactc cctatcagtg atagagaacg tatgtcgagt ttatccctat cagtgataga 2881 gaacgtatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag gtaggcgtgt 2941 acggtgggag gcctatataa gcagagctcg tttagtgaac cgtcagatcg caccggttgc 3001 agatatcaac aagtttgtac aaaaaagttg gcatggggag cagcaagagc aagcccaagg 3061 accccagcca gcgccggcgc aggatgtcag caatacaggc cgcctggcca tccggtacag 3121 aatgtattgc caagtacaac ttccacggca ctgccgagca ggacctgccc ttctgcaaag 3181 gagacgtgct caccattgtg gccgtcacca aggaccccaa ctggtacaaa gccaaaaaca 3241 aggtgggccg tgagggcatc atcccagcca actacgtcca gaagcgggag ggcgtgaagg 3301 cgggtaccaa actcagcctc atgccttggt tccacggcaa gatcacacgg gagcaggctg 3361 agcggcttct gtacccgccg gagacaggcc tgttcctggt gcgggagagc accaactacc 3421 ccggagacta cacgctgtgc gtgagctgcg acggcaaggt ggagcactac cgcatcatgt 3481 accatgccag caagctcagc atcgacgagg aggtgtactt tgagaacctc atgcagctgg 3541 tggagcacta cacctcagac gcagatggac tctgtacgcg cctcattaaa ccaaaggtca 3601 tggagggcac agtggcggcc caggatgagt tctaccgcag cggctgggcc ctgaacatga 3661 aggagctgaa gctgctgcag accatcggga agggggagtt cggagacgtg atgctgggcg 3721 attaccgagg gaacaaagtc gccgtcaagt gcattaagaa cgacgccact gcccaggcct 3781 tcctggctga agcctcagtc atgacgcaac tgcggcatag caacctggtg cagctcctgg 3841 gcgtgatcgt ggaggagaag ggcgggctct acatcgtcac tgagtacatg gccaagggga 3901 gccttgtgga ctacctgcgg tctaggggtc ggtcagtgct gggcggagac tgtctcctca 3961 agttctcgct agatgtctgc gaggccatgg aatacctgga gggcaacaat ttcgtgcatc 4021 gagacctggc tgcccgcaat gtgctggtgt ctgaggacaa cgtggccaag gtcagcgact 4081 ttggtctcac caaggaggcg tccagcaccc aggacacggg caagctgcca gtcaagtgga 4141 cagcccctga ggccctgaga gagaagaaat tctccactaa gtctgacgtg tggagtttcg 4201 gaatccttct ctgggaaatc tactcctttg ggcgagtgcc ttatccaaga attcccctga 4261 aggacgtcgt ccctcgggtg gagaagggct acaagatgga tgcccccgac ggctgcccgc 4321 ccgcagtcta tgaagtcatg aagaactgct ggcacctgga cgccgccatg cggccctcct 4381 tcctacagct ccgagagcag cttgagcaca tcaaaaccca cgagctgcac ctggcggccg 4441 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg 4501 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct 4561 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca 4621 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga 4681 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct 4741 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc 4801 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc 4861 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga 4921 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg 4981 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc 5041 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg 5101 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt 5161 aacccaactt tcttgtacaa agtggttgat atcgactaca aggacgacga tgacaagatc 5221 gactacaagg acgacgatga caagatcgac tacaaggacg acgatgacaa tccatcacac 5281 tggcggccgc tctaaatcga tagtttgttt gaatgaggct tcagtacttt acagaatcgt 5341 tgcctgcaca tcttggaaac acttgctggg attacttctt caggttaacc caacagaagg 5401 ctcgagaagg tatattgctg ttgacagtga gcgatctcgc ttgggcgaga gtaagtagtg 5461 aagccacaga tgtacttact ctcgcccaag cgagagtgcc tactgcctcg gaattcaagg 5521 ggctacttta ggagcaatta tcttgtttac taaaactgaa taccttgcta tctctttgat 5581 acatttttac aaagctgaat taaaatggta taaattaaat cacttttttc aattggaaga 5641 ctaatgcggc cggccattac tccgtctcgt gtcttgttgc atatgtctgc tggtttgttt 5701 gatgttgttt gcgggcgggc cctatagtga gtcgtattac ctaggacgcg tggcctccgc 5761 gccgggtttt ggcgcctccc gcgggcgccc ccctcctcac ggcgagcgct gccacgtcag 5821 acgaagggcg cagcgagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct 5881 gctcataaga ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact 5941 tgggtgactc tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt 6001 cccttctcgg cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata 6061 aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct 6121 tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc 6181 tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg 6241 tagtctgggt ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat 6301 ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga 6361 aacaaggtgg ggggcatggt gggcggcaag aacccaaggt cttgaggcct tcgctaatgc 6421 gggaaagctc ttattcgggt gagatgggct ggggcaccat ctggggaccc tgacgtgaag 6481 tttgtcactg actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt 6541 gccgttgggc agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca 6601 cccgttctgt tggcttataa tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct 6661 tttctccgtc gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg 6721 ccggacctct ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt 6781 acctatcttc ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt 6841 gtgttttgtg aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat 6901 tttcagtgtt agactagtaa attgtccgct aaattctggc cgtttttggc ttttttgtta 6961 gacgctagcg gatccataac ttcgtatagt atacattata cgaagttatg ccaccatgtc 7021 taggctggac aagagcaaag tcataaacgg agctctggaa ttactcaatg gtgtcggtat 7081 cgaaggcctg acgacaagga aactcgctca aaagctggga gttgagcagc ctaccctgta 7141 ctggcacgtg aagaacaagc gggccctgct cgatgccctg ccaatcgaga tgctggacag 7201 gcatcatacc cacttctgcc ccctggaagg cgagtcatgg caagactttc tgcggaacaa 7261 cgccaagtca taccgctgtg ctctcctctc acatcgcgac ggggctaaag tgcatctcgg 7321 cacccgccca acagagaaac agtacgaaac cctggaaaat cagctcgcgt tcctgtgtca 7381 gcaaggcttc tccctggaga acgcactgta cgctctgtcc gccgtgggcc actttacact 7441 gggctgcgta ttggaggaac aggagcatca agtagcaaaa gaggaaagag agacacctac 7501 caccgattct atgcccccac ttctgagaca agcaattgag ctgttcgacc ggcagggagc 7561 cgaacctgcc ttccttttcg gcctggaact aatcatatgt ggcctggaga aacagctaaa 7621 gtgcgaaagc ggcgggccga ccgacgccct tgacgatttt gacttagaca tgctcccagc 7681 cgatgccctt gacgattttg accttgacat gctccccggg taaataactt cgtatagtat 7741 acattatacg aagttatgga tccgcggccg caaattccgc ccctctccct cccccccccc 7801 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt 7861 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt 7921 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt 7981 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct 8041 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt 8101 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt 8161 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa 8221 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta 8281 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa 8341 aacacgataa taccatggcc accgagtaca agcccacggt gcgcctcgcc acccgcgacg 8401 acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc 8461 acaccgtcga cccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca 8521 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg 8581 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggctcgcgca 8641 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc 8701 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc 8761 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgctg 8821 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg 8881 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc 8941 gcaagcccgg tgcctgagtt cgcgtctgga acaatcaacc tctggattac aaaatttgtg 9001 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt 9061 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata 9121 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg 9181 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc 9241 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct 9301 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt 9361 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg 9421 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc 9481 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct 9541 ccctttgggc cgcctccccg cctggaatta attctgcagt cgagacctag aaaaacatgg 9601 agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca 9661 agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta 9721 caaggcagct gtagatctta gccacttttt aaaagaaaag aggggactgg aagggctaat 9781 tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca 9841 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 9901 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 9961 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct 10021 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggccttgaca 10081 ttgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt 10141 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat 10201 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg 10261 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg 10321 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg 10381 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta 10441 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt 10501 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg 10561 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat 10621 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac 10681 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag 10741 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg 10801 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc 10861 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc 10921 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt 10981 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc 11041 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct 11101 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg 11161 agcttgtata tccattttcg gatctgatca gcacgtgatg aaaaagcctg aactcaccgc 11221 gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct 11281 ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct 11341 gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc 11401 atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattaattc agcgagagcc 11461 tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg 11521 aactgcccgc tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc 11581 ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat 11641 ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg 11701 acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg 11761 actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg 11821 acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat 11881 acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc 11941 gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg gcgtatatgc 12001 tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag 12061 cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta 12121 cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg 12181 atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaatagcac gtgctacgag 12241 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg 12301 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact 12361 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata 12421 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc 12481 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc 12541 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt 12601 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc 12661 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg 12721 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct 12781 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca 12841 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga 12901 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc 12961 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg 13021 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat 13081 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt 13141 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc 13201 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg 13261 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg 13321 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg 13381 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg 13441 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca 13501 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 13561 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 13621 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 13681 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 13741 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 13801 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 13861 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 13921 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 13981 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 14041 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 14101 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 14161 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 14221 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 14281 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 14341 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 14401 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 14461 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 14521 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 14581 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 14641 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc 14701 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 14761 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 14821 tatcatgtct ggatcaactg gataactcaa gctaaccaaa atcatcccaa acttcccacc 14881 ccatacccta ttaccactgc caattacctg tggtttcatt tactctaaac ctgtgattcc 14941 tctgaattat tttcatttta aagaaattgt atttgttaaa tatgtactac aaacttagta 15001 gt //