LOCUS Exported 11936 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION lentiviral expression of pHoenix (Synaptophysin-pHluorin-Arch3-mKate2) driven by human Synapsin I promoter for optogenetic acidification of synaptic vesicles. ACCESSION . VERSION . KEYWORDS pLenti-Synapsin-pHoenix-WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11936) AUTHORS Rost BR, Schneider F, Grauel MK, Wozny C, G Bentz C, Blessing A, Rosenmund T, Jentsch TJ, Schmitz D, Hegemann P, Rosenmund C TITLE Optogenetic acidification of synaptic vesicles and lysosomes. JOURNAL Nat Neurosci. 2015 Nov 9. doi: 10.1038/nn.4161. PUBMED 26551543 REFERENCE 2 (bases 1 to 11936) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11936 /organism="synthetic DNA construct" /mol_type="other DNA" regulatory 201..210 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 762..771 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 768..1484 /codon_start=1 /product="pH-sensitive mutant of green fluorescent protein (Ng et al., 2002)" /label=superecliptic pHluorin /note="excited strongly by blue light at neutral pH but minimally fluorescent at acidic pH" /translation="MGSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLL EFVTAAGITHGMDELYK" primer_bind complement(801..829) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 1424..1444 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" CDS 1500..2273 /codon_start=1 /product="light-driven proton pump archaerhodopsin-3 from Halorubrum sodomense (Chow et al., 2010)" /label=Arch /translation="MDPIALQAGYDLLGDGRPETLWLGIGTLLMLIGTFYFLVRGWGVT DKDAREYYAVTILVPGIASAAYLSMFFGIGLTEVTVGGEMLDIYYARYADWLFTTPLLL LDLALLAKVDRVTIGTLVGVDALMIVTGLIGALSHTAIARYSWWLFSTICMIVVLYFLA TSLRSAAKERGPEVASTFNTLTALVLVLWTAYPILWIIGTEGAGVVGLGIETLLFMVLD VTAKVGFGFILLRSRAILGDTEAPEPSAGADVSAAD" CDS 2280..2975 /codon_start=1 /product="monomeric far-red fluorescent protein (Shcherbo et al., 2009)" /label=mKate2 /note="mammalian codon-optimized" /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY CDLPSKLGHR" misc_feature 3747..4335 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3800..3820) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4218..4229) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(4338..4354) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4339..4355) /label=pBluescriptKS /note="For pBluescript vector" LTR 4860..5040 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(5066..5083) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5072..5296 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 5342..5770 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5429..5448) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5639..5660 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5779..5799) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 5784..6113 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5964..6099 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6026..6045 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 6161..6208 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 6227..6601 /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" polyA_signal 6731..6852 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6768..6787) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6822..6841 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6901..6917) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6901..6917) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6914..6936) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6925..6941 /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(6949..6979) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6994..7015 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7132..7149) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7303..7891) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7383..7402) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8062..8922) /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 8685..8704 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8923..9027) /gene="bla" /label=AmpR promoter primer_bind complement(9102..9121) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 9293..9672 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 9674..9872 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 9827..9847 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 9890..10070 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 10117..10242 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 10735..10968 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 11153..11197 /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 11495..11612 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 11681..192 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" ORIGIN 1 ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc cgccggccca 61 gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc 121 ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc 181 ctgagagcgc agtcgctagc gccaccatgg acgtggtgaa tcagctggtg gctgggggtc 241 agttccgggt ggtcaaggag ccccttggct tcgtgaaggt gctgcagtgg gtctttgcca 301 tcttcgcctt tgctacgtgc ggcagctaca ccggagagct tcggctgagc gtggagtgtg 361 ccaacaagac ggagagtgcc ctcaacatcg aagtcgaatt tgagtaccca ttcaggctgc 421 accaagtgta ctttgatgca ccctcctgcg ttaaaggggg cactaccaag atcttcctag 481 ttggtgacta ctcctcctcg gctgaattct ttgtcaccgt ggctgtgttt gccttcctct 541 actccatggg ggccctggcc acctacatct tcctgcagaa caagtaccga gagaacaaca 601 aagggccaat gatggacttc ctggccacag cagtgttcgc tttcatgtgg ctagttagct 661 catccgcctg ggccaaaggc ctgtccgatg tgaagatggc cactgaccca gagaacatta 721 tcaaggagat gcctatgtgc cgccagacag atatggatct agccaccatg ggaagtaaag 781 gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt gatgttaatg 841 ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc aacatacgga aaacttaccc 901 ttaaatttat ttgcactact ggaaaactac ctgttccttg gccaacactt gtcactactt 961 taacttatgg tgttcaatgc ttttcaagat acccagatca tatgaaacgg catgactttt 1021 tcaagagtgc catgcccgaa ggttatgtac aggaaagaac tatatttttc aaagatgacg 1081 ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg 1141 agttaaaagg tattgatttt aaagaagatg gaaacattct tggacacaaa ttggaataca 1201 actataacga tcaccaggtg tacatcatgg cagacaaaca aaagaatgga atcaaagcta 1261 acttcaaaat tagacacaac attgaagatg gaggcgttca actagcagac cattatcaac 1321 aaaatactcc aattggcgat gggcccgtcc ttttaccaga caaccattac ctgtttacaa 1381 cttctactct ttcgaaagat cccaacgaaa agagagacca catggtcctt cttgagtttg 1441 taacagctgc tgggattaca catggcatgg atgaactata caaaaccggt accggatcca 1501 tggaccccat cgctctgcag gctggttacg acctgctggg tgacggcaga cctgaaactc 1561 tgtggctggg catcggcact ctgctgatgc tgattggaac cttctacttt ctggtccgcg 1621 gatggggagt caccgataag gatgcccggg aatattacgc tgtgactatc ctggtgcccg 1681 gaatcgcatc cgccgcatat ctgtctatgt tctttggtat cgggcttact gaggtgaccg 1741 tcgggggcga aatgttggat atctattatg ccaggtacgc cgactggctg tttaccaccc 1801 cacttctgct gctggatctg gcccttctcg ctaaggtgga tcgggtgacc atcggcaccc 1861 tggtgggtgt ggacgccctg atgatcgtca ctggcctcat cggagccttg agccacacgg 1921 ccatagccag atacagttgg tggttgttct ctacaatttg catgatagtg gtgctctatt 1981 ttctggctac atccctgcga tctgctgcaa aggagcgggg ccccgaggtg gcatctacct 2041 ttaacaccct gacagctctg gtcttggtgc tgtggaccgc ttaccctatc ctgtggatca 2101 taggcactga gggcgctggc gtggtgggcc tgggcatcga aactctgctg tttatggtgt 2161 tggacgtgac tgccaaggtc ggctttggct ttatcctgtt gagatcccgg gctattctgg 2221 gcgacaccga ggcaccagaa cccagtgccg gtgccgatgt cagtgccgcc gaccgtacga 2281 tggtgagcga gctgattaag gagaacatgc acatgaagct gtacatggag ggcaccgtga 2341 acaaccacca cttcaagtgc acatccgagg gcgaaggcaa gccctacgag ggcacccaga 2401 ccatgagaat caaggcggtc gagggcggcc ctctcccctt cgccttcgac atcctggcta 2461 ccagcttcat gtacggcagc aaaaccttca tcaaccacac ccagggcatc cccgacttct 2521 ttaagcagtc cttccccgag ggcttcacat gggagagagt caccacatac gaagacgggg 2581 gcgtgctgac cgctacccag gacaccagcc tccaggacgg ctgcctcatc tacaacgtca 2641 agatcagagg ggtgaacttc ccatccaacg gccctgtgat gcagaagaaa acactcggct 2701 gggaggcctc caccgagacc ctgtaccccg ctgacggcgg cctggaaggc agagccgaca 2761 tggccctgaa gctcgtgggc gggggccacc tgatctgcaa cttgaagacc acatacagat 2821 ccaagaaacc cgctaagaac ctcaagatgc ccggcgtcta ctatgtggac agaagactgg 2881 aaagaatcaa ggaggccgac aaagagacct acgtcgagca gcacgaggtg gctgtggcca 2941 gatactgcga cctccctagc aaactggggc acagaaagct tgcagccctg caggagaaga 3001 agtcatgcag ccagcgcatg gccgaattcc ggcaatactg ttggaacccg gacactgggc 3061 agatgctggg ccgcacccca gcccggtggg tgtggatcag cctgtactat gcagctttct 3121 acgtggtcat gactgggctc tttgccttgt gcatctatgt gctgatgcag accattgatc 3181 cctacacccc cgactaccag gaccagttaa agtcaccggg ggtaaccttg agaccggatg 3241 tgtatgggga aagagggctg cagatttcct acaacatctc tgaaaacagc tctggtttaa 3301 acaccaccgg tggagctcga atcacatgca aggaactgag ggaccctgtg acttcaggac 3361 tcaacacctc ggtggtgttt ggcttcctga acctggtgct ctgggttggc aacctatggt 3421 tcgtgttcaa ggagacaggc tgggccgccc cattcatgcg cgcacctcca ggcgccccag 3481 aaaagcaacc agctcctggc gatgcctacg gcgatgcggg ctatgggcag ggccccggag 3541 gctatgggcc ccaggactcc tacgggcctc agggtggtta tcaacccgat tacgggcagc 3601 cagccagcgg cggtggcggt ggctacgggc ctcagggcga ctatgggcag caaggctacg 3661 gccaacaggg tgcgcccacc tccttctcca atcagatgta attaattaag gccgcgactc 3721 tagaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat 3781 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc 3841 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct 3901 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca 3961 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt 4021 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg 4081 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga 4141 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt 4201 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc 4261 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt 4321 gggccgcctc cccgcatcga taccgtcgac ctcgagacct agaaaaacat ggagcaatca 4381 caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg 4441 aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag 4501 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc 4561 aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt 4621 ggcagaacta cacaccaggg ccagggatca gatatccact gacctttgga tggtgctaca 4681 agctagtacc agttgagcaa gagaaggtag aagaagccaa tgaaggagag aacacccgct 4741 tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagta ttagagtgga 4801 ggtttgacag ccgcctagca tttcatcaca tggcccgaga gctgcatccg gactgtactg 4861 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac 4921 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt 4981 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca 5041 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt 5101 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc 5161 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt 5221 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat 5281 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc 5341 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc 5401 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc 5461 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt 5521 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg 5581 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt 5641 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta 5701 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt 5761 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc 5821 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt 5881 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 5941 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc 6001 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg 6061 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc 6121 cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc 6181 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac 6241 cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga 6301 ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga 6361 cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg 6421 ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa 6481 cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga 6541 gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg 6601 acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat 6661 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt 6721 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 6781 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 6841 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg 6901 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 6961 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 7021 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 7081 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 7141 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 7201 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 7261 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 7321 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 7381 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 7441 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 7501 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 7561 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 7621 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 7681 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 7741 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 7801 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 7861 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 7921 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 7981 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 8041 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 8101 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 8161 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 8221 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 8281 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 8341 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 8401 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 8461 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 8521 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 8581 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 8641 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 8701 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 8761 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 8821 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 8881 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 8941 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 9001 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga 9061 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc 9121 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg 9181 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt 9241 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga 9301 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg 9361 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc 9421 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat 9481 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat 9541 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat 9601 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc 9661 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac 9721 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa 9781 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt 9841 aggcgtgtac ggtgggaggt ctatataagc agcgcgtttt gcctgtactg ggtctctctg 9901 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc 9961 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg 10021 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg 10081 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt 10141 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg 10201 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga 10261 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta 10321 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta 10381 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga 10441 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg 10501 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt 10561 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga 10621 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc 10681 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc 10741 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct 10801 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag 10861 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca 10921 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg 10981 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa 11041 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa 11101 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga 11161 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa 11221 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat 11281 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt 11341 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg 11401 tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac tgcgtgcgcc 11461 aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg 11521 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat 11581 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc 11641 cagtttggtt aatcgcatgc tagagagggc cctgcgtatg agtgcaagtg ggttttagga 11701 ccaggatgag gcggggtggg ggtgcctacc tgacgaccga ccccgaccca ctggacaagc 11761 acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg ggaaacagga 11821 tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc 11881 ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca ctcgcc //