LOCUS Exported 6901 bp ds-DNA circular SYN 14-JUL-2018 DEFINITION . ACCESSION . VERSION . KEYWORDS pAAV-hSyn-LMO4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6901) AUTHORS Hochgeschwender TITLE Direct Submission JOURNAL Exported Jul 21, 2018 from SnapGene 4.1.1 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6901 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 5..145 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" /note="color: #ffe4c4" promoter 190..637 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" /note="color: #ffffff; direction: RIGHT" regulatory 678..687 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /note="color: #a6acb3" CDS 684..1238 /codon_start=1 /product="enhanced variant of secreted Gaussia luciferase" /label=Gaussia luciferase /note="human codon-optimized" /note="This feature has 2 segments: 1: 684 .. 734 / #ff0000 / signal sequence 2: 735 .. 1238 / #00ccff / GLucM23" /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG KLPGEKLPLEVLKELEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA QGGIGEAIDDIPEIPGFKDLEPIEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ RCATFASKIQGQVDKIKGAGDD" CDS 1239..1283 /codon_start=1 /label=15 aa Linker /note="color: #993300" /translation="GDPLVQCGGIAGSAT" CDS 1284..2183 /codon_start=1 /label=VChR1 /note="color: #993366" /translation="MDYPVARSLIVRYPTDLGNGTVCMPRGQCYCEGWLRSRGTSIEKT IAITLQWVVFALSVACLGWYAYQAWRATCGWEEVYVALIEMMKSIIEAFHEFDSPATLW LSSGNGVVWMRYGEWLLTCPVLLIHLSNLTGLKDDYSKRTMGLLVSDVGCIVWGATSAM CTGWTKILFFLISLSYGMYTYFHAAKVYIEAFHTVPKGICRELVRVMAWTFFVAWGMFP VLFLLGTEGFGHISPYGSAIGHSILDLIAKNMWGVLGNYLRVKIHEHILLYGDIRKKQK ITIAGQEMEVETLVAEEED" CDS 2193..2909 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /note="This feature has 2 segments: 1: 2193 .. 2195 / #e1ff2f / 1a 2: 2196 .. 2909 / #e1ff2f" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" CDS 2933..2974 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /note="color: #cc99b2" /translation="GKPIPNPLLGLDST" CDS 2984..3001 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /note="color: #cc99b2" /translation="HHHHHH" misc_feature 3032..3620 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" /note="color: #ffffff" polyA_signal 3652..4128 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" /note="color: #a6acb3" repeat_region 4168..4308 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" /note="color: #ffe4c4" rep_origin 4383..4838 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /note="color: #ffff00" promoter 5120..5224 /gene="bla" /label=AmpR promoter /note="color: #ffffff; direction: RIGHT" CDS 5225..6085 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /note="This feature has 2 segments: 1: 5225 .. 5293 / #ccffcc / signal sequence 2: 5294 .. 6085 / #ccffcc Cleavage site after base 5293" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 6256..6844 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" ORIGIN 1 atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg 61 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg 121 gccaactcca tcactagggg ttcctgcggc cgcacgcgtg tgtctagact gcagagggcc 181 ctgcgtatga gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct 241 gacgaccgac cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc 301 cctatcagag agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc 361 agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac 421 tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg 481 tcgcgtccgc gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg 541 gcacgggcgc gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt 601 gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg agaaggtacc ggatcctcta 661 gagtcgagct caagcttgcc accatgggag tcaaagttct gtttgccctg atctgcatcg 721 ctgtggccga ggccaagccc accgagaaca acgaagactt caacatcgtg gccgtggcca 781 gcaacttcgc gaccacggat ctcgatgctg accgcgggaa gttgcccggc gagaagctgc 841 cgctggaggt gctcaaagag ttggaagcca atgcccggaa agctggctgc accaggggct 901 gtctgatctg cctgtcccac atcaagtgca cgcccaagat gaagaagttc atcccaggac 961 gctgccacac ctacgaaggc gacaaagagt ccgcacaggg cggcataggc gaggcgatcg 1021 acgacattcc tgagattcct gggttcaagg acttggagcc catagagcag ttcatcgcac 1081 aggtcgatct gtgtgtggac tgcacaactg gctgcctcaa agggcttgcc aacgtgcagt 1141 gttctgacct gctcaagaag tggctgccgc aacgctgcgc gacctttgcc agcaagatcc 1201 agggccaggt ggacaagatc aagggggccg gtgatgacgg cgacccacta gtccagtgtg 1261 gtggaattgc cggatccgcc accatggatt accctgttgc tagaagcctc atagttcgct 1321 acccaaccga cctcggaaac ggcaccgtct gcatgccaag aggacagtgt tactgtgaag 1381 gttggcttcg gagtcgcggc acttccattg aaaagacaat agcaattact cttcagtggg 1441 tagtctttgc tttgtcagtg gcttgcctgg ggtggtatgc gtatcaagcg tggcgagcta 1501 cctgcggatg ggaggaggtt tacgtagcct tgatagaaat gatgaaaagc atcatcgagg 1561 ccttccacga gttcgacagc cctgcaacac tgtggctgtc ttcagggaac ggcgtagttt 1621 ggatgcggta tggcgaatgg ctcctcacct gcccggtcct tctgatccat ctgagcaacc 1681 tcacaggcct gaaggacgat tatagcaaaa ggactatggg cctgttggtt tctgatgtgg 1741 gatgcatcgt gtggggcgca accagcgcca tgtgtacggg gtggacgaag atcctgttct 1801 tcctcatctc attgagctat ggtatgtata cctattttca tgctgctaaa gtttatatcg 1861 aagcattcca cacagttcca aaagggattt gtcgagaact ggtccgagtg atggcctgga 1921 cattctttgt ggcttgggga atgtttccag tcctgtttct gctgggcacg gaaggattcg 1981 gtcatatcag cccttatgga tctgccattg ggcactccat cctcgacctg attgcaaaga 2041 acatgtgggg tgtgctgggg aattacctgc gcgtcaaaat ccacgagcac atcctgttgt 2101 atggcgacat cagaaagaag cagaaaatta cgatcgccgg ccaagagatg gaggttgaga 2161 cactggtggc tgaagaggag gacgcggccg ccgtgagcaa gggcgaggag ctgttcaccg 2221 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt 2281 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca 2341 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcctgcagt 2401 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg 2461 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg 2521 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact 2581 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg 2641 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca 2701 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg 2761 acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag 2821 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca 2881 ctctcggcat ggacgagctg tacaagtaac tctagagggc ccgcggttcg aaggtaagcc 2941 tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc accatcacca 3001 ttgagtttaa ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga 3061 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt 3121 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa 3181 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt 3241 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct 3301 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg 3361 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc 3421 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg 3481 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct 3541 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc 3601 cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat 3661 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc 3721 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat 3781 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg 3841 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca 3901 atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc 3961 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc 4021 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc 4081 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag 4141 gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct 4201 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct 4261 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc 4321 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata 4381 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac 4441 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc 4501 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt 4561 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg 4621 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag 4681 tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt 4741 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt 4801 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat 4861 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc 4921 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag 4981 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt 5041 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg 5101 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa 5161 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa 5221 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct 5281 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg 5341 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg 5401 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt 5461 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga 5521 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga 5581 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac 5641 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg 5701 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac 5761 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct 5821 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct 5881 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg 5941 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat 6001 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg 6061 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat 6121 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct 6181 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa 6241 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa 6301 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc 6361 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta 6421 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct 6481 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg 6541 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag 6601 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc 6661 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg 6721 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt 6781 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg 6841 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca 6901 c //