LOCUS Exported 6854 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION An AAV packaging vector that expresses eGFP and Kv1.2 under the control of a neuronal promoter.. ACCESSION . VERSION . KEYWORDS pOTTC533 - pAAV SYN1 eGFP-2A-mKv1.2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6854) AUTHORS Kourrich et al. TITLE AAV Potassium channel JOURNAL Unpublished REFERENCE 2 (bases 1 to 6854) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6854 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..141 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 12..141 /label=AAV2 ITR promoter 186..633 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" regulatory 653..662 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 659..1375 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 659..1375 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(704..725) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(965..984) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1312..1333 /label=EGFP-C /note="EGFP, forward primer" CDS 1397..1450 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" misc_feature 2981..3569 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3034..3054) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3452..3463) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3601..4077 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3800..3819) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4117..4257 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4117..4246 /label=AAV2 ITR rep_origin 4332..4787 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4419..4438) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4629..4650 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4804..4823) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4923..4945 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4983..5001) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5069..5173 /gene="bla" /label=AmpR promoter CDS 5174..6034 /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 complement(5392..5411) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6205..6793 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6694..6713 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 121 actccatcac taggggttcc tgcggccgca cgcgtgtgtc tagactgcag agggccctgc 181 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg 241 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta 301 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca 361 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa 421 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc 481 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac 541 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc 601 agcggaggag tcgtgtcgtg cctgagagcg cagtcgagaa ggtaccggat ccgccaccat 661 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg 721 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg 781 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct 841 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca 901 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt 961 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt 1021 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa 1081 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg 1141 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga 1201 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta 1261 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct 1321 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagatgca 1381 ttctagaggc tccggagagg gcaggggaag tcttctaaca tgcggggacg tggaggaaaa 1441 tcccgggccc atcgatatga cagtggctac cggagaccca gtggacgagg ctgctgccct 1501 ccctgggcac ccacaagaca cctatgaccc agaggcagac catgaatgct gtgagagagt 1561 ggtcatcaac atctcaggcc tgcggttcga aactcagcta aagaccttag cccagttccc 1621 agagaccctc ttaggggacc ccaagaaacg gatgaggtac ttcgatcccc tccgaaatga 1681 gtactttttt gatcgcaacc gccctagctt tgatgccatt ttgtactact atcagtctgg 1741 gggcaggttg aggcgacctg tgaacgtgcc cttagatatc ttctcggaag aaatccggtt 1801 ttatgagcta ggagaagaag caatggagat gtttcgggag gatgaaggct acatcaagga 1861 agaagagcgt cctctgcctg aaaatgagtt tcagagacag gtgtggcttc tctttgaata 1921 ccctgagagc tcagggcctg ccaggattat agccattgta tctgtgatgg tcattctgat 1981 ctccatcgtc agcttctgtc tggaaacctt gcccatcttc cgggatgaga atgaggacat 2041 gcatggtggc ggggtgacct tccacaccta ttccaacagc accatcgggt accagcagtc 2101 cacctccttc accgaccctt tcttcattgt agagactctc tgcatcatct ggttctcctt 2161 tgagtttctg gttagattct ttgcctgtcc cagcaaagct ggcttcttca ccaacatcat 2221 gaacatcatt gacattgtgg ctatcatccc ttactttatc accctgggga cagagttagc 2281 tgagaagcca gaggatgccc agcaaggcca gcaggccatg tcactggcca ttctccgtgt 2341 catccggttg gtaagagtct ttaggatttt caagttgtcc agacactcca aaggtctaca 2401 gattctaggt cagaccctca aagctagcat gagggaattg ggcctcctga tattcttcct 2461 cttcattggg gtcatcctct tctctagtgc tgtctatttt gcagaagctg atgagagaga 2521 ttcccagttc cccagcatcc cggatgcttt ctggtgggca gtcgtctcca tgacaactgt 2581 aggctatgga gacatggttc caactaccat tggggggaag atagtgggtt ctctgtgtgc 2641 aattgcaggt gtgttaacca ttgccttacc agtccctgtc atagtgtcta atttcaacta 2701 cttctaccac cgggagacag agggagagga gcaggcccag tacttgcaag tgacaagctg 2761 tccaaagatc ccgtcctccc ctgacctaaa gaaaagtaga agtgcctcta ccataagtaa 2821 gtctgattac atggagatac aggagggagt taacaacagc aatgaggact ttagagagga 2881 gaacttaaaa acagccaact gtaccttggc taacacaaac tatgtgaata ttaccaaaat 2941 gttaactgat gtctgagaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa 3001 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata 3061 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc 3121 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg 3181 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac 3241 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat 3301 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt 3361 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat 3421 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc 3481 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag 3541 tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg agagatctac 3601 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca 3661 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc 3721 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca 3781 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg 3841 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg 3901 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg 3961 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct 4021 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg 4081 attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg 4141 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac 4201 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc 4261 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa 4321 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 4381 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 4441 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 4501 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc 4561 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 4621 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc 4681 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 4741 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct 4801 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc 4861 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt 4921 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa 4981 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac 5041 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat 5101 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg 5161 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc 5221 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga 5281 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga 5341 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg 5401 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc 5461 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac 5521 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact 5581 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca 5641 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg 5701 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact 5761 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg 5821 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg 5881 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat 5941 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc 6001 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat 6061 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt 6121 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc 6181 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt 6241 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac 6301 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt 6361 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct 6421 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga 6481 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac 6541 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg 6601 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt 6661 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc 6721 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg 6781 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc 6841 ttttgctcac atgt //