LOCUS Exported 14389 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION expresses mouse Piezo1 with GFP. ACCESSION . VERSION . KEYWORDS mPiezo1-IRES-eGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 14389) AUTHORS Coste B, Mathur J, Schmidt M, Earley TJ, Ranade S, Petrus MJ, Dubin AE, Patapoutian A TITLE Piezo1 and Piezo2 are essential components of distinct mechanically activated cation channels. JOURNAL Science. 2010 Oct 1;330(6000):55-60. doi: 10.1126/science.1193270. Epub 2010 Sep 2. PUBMED 20813920 REFERENCE 2 (bases 1 to 14389) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..14389 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 124..503 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 504..707 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 658..678 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 752..771 /label=T7 /note="T7 promoter, forward primer" promoter 752..770 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 823..832 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" misc_feature 8528..9114 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(8695..8712) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 8922..8941 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 9115..9834 /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 9115..9831 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(9160..9181) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(9421..9440) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 9768..9789 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(9869..9886) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 9875..10099 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 10145..10573 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(10232..10251) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 10442..10463 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(10582..10602) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 10587..10916 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 10767..10902 /label=SV40 ori /note="SV40 origin of replication" primer_bind 10829..10848 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 10983..11777 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(11037..11056) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 11647..11666 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 11951..12072 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(11988..12007) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 12042..12061 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(12121..12137) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(12121..12137) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(12134..12156) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 12145..12161 /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(12169..12199) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 12214..12235 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(12352..12369) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(12523..13111) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(12603..12622) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(13282..14142) /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 13905..13924 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(14143..14247) /gene="bla" /label=AmpR promoter primer_bind complement(14322..14341) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga 61 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc 121 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 181 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 241 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 301 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 361 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 421 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 481 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat 541 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt 601 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc 661 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta 721 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag 781 ctggctagcg tttaaacggg ccctctagac tcgagcggcc gcgccaccat ggagccgcac 841 gtgctgggcg ccgggctcta ctggctgttg ctgccctgca cgctcctggc ggcctccctg 901 ttacgcttca atgctctctc gctggtctat ttgctgtttc tactgctgct gccctggctt 961 ccaggcccct caagacacag cataccaggt cacacaggtc gcctgctccg tgcactactc 1021 tgcctcagcc tcctcttcct ggtggcccac cttgcctttc agatatgcct acacaccgtg 1081 cctcacctgg accagtttct gggacaaaac ggtagccttt gggtgaaggt gtctcaacac 1141 ataggggtta cgaggctgga cctgaaggac atctttaaca ccaccaggct ggtagcacct 1201 gacctgggag tgctgctggc gtcctccctt tgccttggcc tctgtggacg cctcacgagg 1261 aaagccgggc agagtcggcg cacccaggag ctgcaggatg atgacgatga cgatgatgac 1321 gacgatgaag acatagatgc tgccccagcc gtggggctga agggagcccc tgccctggca 1381 accaaacgca ggctgtggct ggcctcccgc ttccgggtca cggcccactg gctgctgatg 1441 acctctggac ggacgctggt cattgtgttg ctggccctgg caggcatagc ccacccttcg 1501 gccttctcca gcatctacct ggtggtgttc ctggccatct gcacctggtg gtcctgtcac 1561 tttcctctca gccccctggg ctttaacacc ctctgtgtca tggtgagctg ttttggtgcc 1621 ggccatctca tttgcctata ctgctatcag acaccattta tccaggacat gttaccccct 1681 gggaacatct gggccaggct atttggtctc aagaacttcg tagacctccc taactactcc 1741 agccccaacg ccctggtact caacactaag cacgcctggc ccatctatgt gagtcccgga 1801 atcctgctgc tgctatatta cacagccacc tctctcctga agctccacaa gagctgtccc 1861 tcagagctga ggaaggagac acccagggag gatgaggagc atgagctgga actggaccac 1921 ctggagccag agccccaggc tagggacgcc acccagggtg agatgcccat gaccacggaa 1981 cctgaccttg acaactgcac cgtgcatgta ctaaccagcc agagccctgt ccgccagcgt 2041 ccagttcgcc ccaggctggc tgagctgaaa gagatgtcac cgctacatgg cctgggccac 2101 ctcatcatgg accagagcta tgtgtgtgcc ctgattgcca tgatggtgtg gagcatcatg 2161 taccacagtt ggctgacctt cgtcctgctg ctctgggcct gcctcatctg gacggtgcgt 2221 agccgtcacc agctggctat gctctgctcg ccctgcatcc tgctgtatgg gctgacgctc 2281 tgctgcctga ggtatgtgtg ggccatggaa cttcctgagc tgcccaccac cctgggccct 2341 gtcagcctgc accagttggg actggaacac acacgctacc cttgcctgga cctcggtgcc 2401 atgctcctct atctgctcac attctggctc cttctgcgtc agttcgtgaa ggagaagctg 2461 ctgaagaagc aaaaggtgcc tgcggcgctg ctggaggtca cggtggccga cactgagccc 2521 acacagaccc agacgctgct gcggagcctg ggggagctgg tcaccggcat ctacgtcaaa 2581 tactggatct atgtgtgcgc cggcatgttc attgtggtca gcttcgccgg ccgcctggtg 2641 gtctacaaaa tcgtctacat gttcctcttc ctgctgtgcc tcaccttgtt ccaggtctac 2701 tacaccctgt ggaggaagct gctacgtgtc ttctggtggc tcgtggtggc ctatacaatg 2761 ctcgtgctca tcgctgtgta caccttccag ttccaggact tccccaccta ttggcgcaac 2821 ctcacgggct tcacagacga gcagttgggc gacctgggcc tggagcagtt cagtgtgtcg 2881 gagctctttt ccagtatcct catccctggc ttcttcctgc tggcctgcat cctgcagctg 2941 cactacttcc acagaccgtt catgcagctc actgacctgg agcacgtgcc gccaccaggc 3001 acccgccacc ctcgatgggc tcacaggcag gatgcagtga gcgaggcccc tctgcttgag 3061 catcaggagg aagaggaagt cttcagggaa gatgggcaga gcatggatgg gccccaccag 3121 gccacacagg tccctgaggg tacggccagc aagtggggcc tggtggctga ccggctgctg 3181 gacctagcgg ccagcttctc ggctgtcctc acccgaatcc aggtgttcgt tcggcgcttg 3241 ctagaacttc acgtcttcaa gctggtggcc ctctacactg tctgggtggc cctgaaagaa 3301 gtgtctgtga tgaacctgct gctggtggtg ctatgggcct tcgccttgcc ctatccgcgc 3361 ttccggccca tggcttcctg cctgtccacc gtgtggacct gtatcatcat tgtatgcaag 3421 atgctctatc agctcaagat tgtcaacccg catgagtact ccagcaactg cactgagccc 3481 ttccccaaca ataccaactt gcagcctttg gagatcaacc agtctttgct gtaccgtggc 3541 cctgttgacc ctgccaactg gtttggggtg cggaagggtt accccaactt gggctatatc 3601 cagaaccacc tgcagatcct tctgttgctg gtgtttgagg ccgtggtgta ccggcgccaa 3661 gagcactacc gccggcagca ccagcaggcc cctctgcccg cccaggctgt gtgcgcagat 3721 ggcacccgcc agaggttgga ccaggaccta cttagctgcc tcaagtattt catcaacttc 3781 ttcttctaca aattcgggct ggagatctgc ttcttgatgg ccgtgaatgt gattgggcag 3841 cgtatgaact tcatggtgat cttacacggt tgctggttgg tggccatcct tacacgccgg 3901 cgccgtgagg ccatcgcccg cctctggcct aactactgtc tgttcctcac gctgttcctg 3961 ctgtaccagt acctgctgtg tttgggcatg ccccctgcac tgtgcattga ctatccatgg 4021 cgctggagca aggccatccc catgaattcc gccctcatca agtggctgta cctacctgac 4081 ttcttcagag cccccaactc caccaacctt atcagtgact tcctcctgct gctttgcgcc 4141 tcccagcagt ggcaggtctt ctcagcggag cgaacggagg agtggcaacg catggcgggc 4201 atcaacactg accacctgga gcccctgcgt ggggagccca accctatacc caacttcatc 4261 cactgcaggt cctatctgga tatgctgaag gtggccgtct tccgctacct gttctggctg 4321 gtgctcgttg tggtgtttgt tgcgggggcc acccgcataa gcatcttcgg gctggggtac 4381 ctgctagcct gcttctacct gctgctgttt ggcactaccc tgctgcagaa ggacacgcga 4441 gcccagctcg tgctgtggga ctgcctcatc ctctataatg tcactgtcat catctctaag 4501 aatatgctgt cgctcctgtc ctgtgtcttc gtggagcaaa tgcagagcaa cttctgctgg 4561 gtcatccagc tcttcagcct cgtgtgcaca gtcaaaggct actatgatcc caaagagatg 4621 atgaccaggg accgggactg cctgctgcct gtggaggagg ccgggatcat ctgggacagt 4681 atctgcttct tcttcctgct cttgcaacgg cgcatctttc tcagccacta cttcctgcat 4741 gtcagcgctg acctgaaagc cacagccctg caggcatcca ggggctttgc cctctacaat 4801 gcagccaacc tgaagagcat caacttccat cgccagattg aggagaagtc cctggcccag 4861 ctgaaaagac agatgaagcg catccgtgcc aaacaggaga agtacaggca gagccaggca 4921 agtcgtggcc aactccagtc caaagaccct caggatccca gccaggagcc agggcctgac 4981 agcccagggg gctcctcccc gccacggaga cagtggtggc gcccctggct ggaccacgcc 5041 acagtcatcc actctggcga ctacttcctg tttgagtcag atagcgagga ggaagaggag 5101 gccctacctg aggaccccag gcctgcagct cagagtgcct tccagatggc ataccaggca 5161 tgggtaacca atgcccagac agtgctgagg cagcgtcggg agcgggcacg gcaggagcgg 5221 gcagagcagc tggcttctgg aggtgacttg aacccagatg tggaaccagt agatgtccca 5281 gaagatgaga tggcaggccg tagccacatg atgcagcgtg tgctaagcac catgcagttc 5341 ctgtgggtgc tgggccaggc cacggtagac gggctgacgc gctggctgcg tgcattcacg 5401 aagcaccacc gcaccatgag cgatgtgctg tgcgcagagc gctacctgct cacccaggag 5461 cttcttcggg ttggagaggt acgccgaggt gtgctggacc agctttatgt gggtgaagat 5521 gaggccacat tgtcaggtcc cgtggagacc cgggatggac ccagcacagc ctcaagtggg 5581 ctgggagccg aagagccttt gagtagcatg acagacgaca ccagcagccc cctgagcaca 5641 ggctataaca cccgcagtgg cagtgaggag attgtcaccg acgctgggga cctccaggct 5701 gggacctccc tgcacggctc ccaagagctt ttagccaatg ctcgtacccg gatgcgcacg 5761 gccagcgagc tgctactgga taggcgcctg catatccctg agctggagga ggccgagcgg 5821 tttgaggcac agcagggccg gactctgcgg ctgctcaggg ctgggtacca gtgcgtggcg 5881 gcacactcgg agctgctctg ttacttcatc atcatcctta accacatggt gacagcctcg 5941 gctgcctccc tggtgctgcc cgtgcttgtg ttcctgtggg ccatgctgac catcccgagg 6001 cctagcaagc gcttttggat gacagctatc gtcttcactg aggtcatggt ggtcaccaaa 6061 tacctgttcc agttcggctt cttcccctgg aacagctacg ttgtgctgcg gcgctatgag 6121 aacaagccct acttccctcc gcgaatcctg ggccttgaga aaacggacag ctacatcaag 6181 tatgacctgg tgcagctcat ggccctcttc ttccaccgct cgcagctact gtgttatggc 6241 ctctgggacc atgaggagga tcgctatccc aaggaccatt gcaggagtag tgtgaaggac 6301 cgggaggcca aggaagagcc agaagctaag ctggaatcgc agtctgagac aggcactggg 6361 catcccaagg agccagtgtt ggccggtact cccagggacc acatccaagg gaaaggaagt 6421 attagatcca aggatgttat ccaagatccc ccagaggacc ttaagccccg gcacacgagg 6481 cacatcagca tacgcttcag gaggcgcaag gagactccag gacccaaagg aacagcagtc 6541 atggagactg agcacgagga gggagaagga aaagaaacta cagagagaaa gaggccgcgt 6601 cacactcaag aaaaatcgaa gtttcgggag agaatgaagg cagctgggcg ccggctgcag 6661 agcttctgtg tgtcactggc ccagagcttc taccaaccct tgcagcgttt cttccatgac 6721 attctgcaca caaagtaccg ggcggccacc gacgtctacg ccctcatgtt cctggccgat 6781 attgtcgaca tcatcatcat catctttggt ttttgggctt ttgggaagca ctctgcagcc 6841 acagacattg catcctcgct gtcagatgac caggtgccac aggccttcct gttcatgctg 6901 ctggtccagt ttggcaccat ggtcatcgac cgtgccctct acctgcgcaa gactgtcctg 6961 ggaaagctgg cctttcaggt ggtcctggtg gtggcgattc acatctggat gttctttatc 7021 ttaccggctg tcactgagag gatgttcagc cagaatgcgg tggcacagct gtggtacttc 7081 gtcaagtgca tttactttgc cctgtccgcc taccagatcc gctgtggcta ccccacccgt 7141 atcttgggca acttcctcac caagaaatac aaccatctaa acctcttcct tttccagggg 7201 ttccgtctag tgccgttcct ggtggagctg cgggccgtca tggactgggt gtggaccgac 7261 accacgctgt ccctgtccaa ctggatgtgt gtggaagaca tctatgccaa catctttatc 7321 atcaagtgca gccgagagac agagaagaaa tacccccagc ccaaggggca gaagaagaag 7381 aaaattgtca agtatggtat gggaggcctc attatcctct tcctcatcgc catcatctgg 7441 ttccctctgc tcttcatgtc actgatccgc tctgtggtcg gggtcgtcaa ccagcccatt 7501 gatgtcaccg tcaccctcaa gctaggcggc tatgagccac tgttcaccat gagcgcccag 7561 cagccatcca ttgtgccatt cacaccccag gcctacgagg agctgtccca gcagtttgac 7621 ccctatccac tagccatgca gttcattagc cagtacagtc ctgaggacat cgtcactgca 7681 cagatcgagg gcagctcggg ggcgctgtgg cgcatcagcc cacccagccg agcccagatg 7741 aagcaggagc tgtacaacgg cacagccgac attacactgc gctttacctg gaatttccaa 7801 agggacctgg ccaagggtgg cactgtggag tatactaatg agaagcacac cttggagctg 7861 gcccccaaca gtacggcacg aaggcagctg gcccaactgc tcgagggcag acctgaccag 7921 tcagtggtca ttccccatct cttccccaag tacattcgtg ctcccaatgg gcctgaagcc 7981 aaccctgtga agcagctgca gccagatgag gaagaggact accttggtgt gcgcatccag 8041 ctgcggaggg agcaagtggg cacaggggcc tctggggagc aagcgggcac caaggcctcc 8101 gacttcctcg agtggtgggt catcgagctg caggactgca aggctgactg caacctgctg 8161 cccatggtca tcttcagtga caaggtcagc ccacctagcc tgggcttcct ggccggctac 8221 gggattgtgg ggctgtacgt ctccatcgtg ctggtggttg gcaagtttgt gcggggcttc 8281 ttcagcgaga tctctcactc catcatgttc gaggaactgc cgtgtgtgga ccgcatcctc 8341 aagctgtgcc aggacatctt cttggtgcgc gagacccggg agctggagct ggaggaggag 8401 ctatacgcca agctcatctt cctgtaccga tctccagaga ccatgattaa gtggacacgt 8461 gagagggagt agaagcttgg cgcgccttaa ttaactgcag tcgacggtac cgcgggcccg 8521 ggatccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag 8581 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga 8641 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg 8701 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt 8761 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca 8821 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc 8881 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat 8941 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc 9001 ctcggtacac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga 9061 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatggtg 9121 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac 9181 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag 9241 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg 9301 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac 9361 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag 9421 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac 9481 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg 9541 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc 9601 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac 9661 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg 9721 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg 9781 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaggccgg 9841 ccaagcttaa gtttaaaccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct 9901 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt 9961 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg 10021 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg 10081 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat 10141 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 10201 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 10261 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga 10321 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt 10381 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 10441 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat 10501 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 10561 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct 10621 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa 10681 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 10741 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 10801 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct 10861 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc 10921 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc 10981 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat 11041 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt 11101 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac 11161 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg 11221 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc 11281 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa 11341 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc 11401 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg 11461 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg 11521 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa 11581 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg 11641 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct 11701 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc 11761 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa 11821 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat 11881 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt 11941 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 12001 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 12061 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg 12121 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 12181 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 12241 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 12301 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 12361 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 12421 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 12481 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 12541 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 12601 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 12661 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 12721 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 12781 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 12841 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 12901 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 12961 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 13021 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 13081 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 13141 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 13201 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 13261 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 13321 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 13381 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 13441 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 13501 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 13561 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 13621 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 13681 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 13741 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 13801 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 13861 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 13921 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 13981 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 14041 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 14101 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 14161 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 14221 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga 14281 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc 14341 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctg //