LOCUS Exported 7080 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Optogenetics. ACCESSION . VERSION . KEYWORDS pAAV-hSyn-GCaMP 6m-p2A-ChRmine-Kv2.1-WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7080) AUTHORS Marshel JH, Kim YS, Machado TA, Quirin S, Benson B, Kadmon J, Raja C, Chibukhchyan A, Ramakrishnan C, Inoue M, Shane JC, McKnight DJ, Yoshizawa S, Kato HE, Ganguli S, Deisseroth K TITLE Cortical layer-specific critical dynamics triggering perception. JOURNAL Science. 2019 Aug 9;365(6453). pii: science.aaw5202. doi: 10.1126/science.aaw5202. Epub 2019 Jul 18. PUBMED 31320556 REFERENCE 2 (bases 1 to 7080) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7080 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 6..135 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 180..627 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" regulatory 641..650 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 662..718 /codon_start=1 /product="calmodulin-binding peptide from avian smooth muscle myosin light chain kinase" /label=calmodulin-binding peptide /translation="RRKWNKTGHAVRAIGRLSS" primer_bind 931..952 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1058..1079) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1319..1338) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" CDS 1910..1966 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 3158..3184 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" misc_feature 3212..3800 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3265..3285) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3683..3694) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3832..4308 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4031..4050) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4348..4488 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4348..4477 /label=AAV2 ITR rep_origin 4563..5018 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4650..4669) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4860..4881 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5035..5054) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5154..5176 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5214..5232) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5300..5404 /gene="bla" /label=AmpR promoter CDS 5405..6265 /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(5623..5642) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6436..7024 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6925..6944 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga 61 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca 121 tcactagggg ttcctgcggc cgcacgcgtg tgtctagact gcagagggcc ctgcgtatga 181 gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct gacgaccgac 241 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag 301 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg 361 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg 421 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc 481 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc 541 gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga 601 ggagtcgtgt cgtgcctgag agcgcagtcg agaaggatcc gccaccatgg tggactcatc 661 acgtcgtaag tggaataaga caggtcacgc agtcagagct ataggtcggc tgagctcact 721 cgagaacgtc tatatcaagg ccgacaagca gaagaacggc atcaaggcga acttcaagat 781 ccgccacaac atcgaggacg gcggcgtgca gctcgcctac cactaccagc agaacacccc 841 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcgtgc agtccaaact 901 ttcgaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc 961 cgggatcact ctcggcatgg acgagctgta caagggcggt accggaggga gcatggtgag 1021 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt 1081 aaacggccac aagttcagcg tgtccggcga gggtgagggc gatgccacct acggcaagct 1141 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac 1201 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga 1261 cttcttcaag tccgccatgc ccgaaggcta catccaggag cgcaccatct tcttcaagga 1321 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg 1381 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga 1441 gtacaacctg ccggaccaac tgactgaaga gcagatcgca gaatttaaag aggctttctc 1501 cctatttgac aaggacgggg atgggacaat aacaaccaag gagctgggga cggtgatgcg 1561 gtctctgggg cagaacccca cagaagcaga gctgcaggac atgatcaatg aagtagatgc 1621 cgacggtgac ggcacaatcg acttccctga gttcctgaca atgatggcaa gaaaagggag 1681 ctacagggac acggaagaag aaattagaga agcgttcggt gtgtttgata aggatggcaa 1741 tggctacatc agtgcagcag agcttcgcca cgtgatgaca aaccttggag agaagttaac 1801 agatgaagag gttgatgaaa tgatcaggga agcagacatc gatggggatg gtcaggtaaa 1861 ctacgaagag tttgtacaaa tgatgacagc gaagactagt ggctccggag ccacgaactt 1921 ctctctgtta aagcaagcag gagacgtgga agaaaacccc ggtcccatgg cacacgcacc 1981 aggcaccgac cagatgttct acgtgggcac aatggacggc tggtatctgg ataccaagct 2041 gaactccgtg gccatcggcg cccactggtc ttgctttatc gtgctgacaa tcaccacatt 2101 ctacctgggc tatgagtcct ggacatctcg gggccctagc aagagaacct ccttttacgc 2161 cggctatcag gaggagcaga acctggccct gtttgtgaat ttctttgcca tgctgagcta 2221 cttcggcaag atcgtggccg acacactggg ccacaacttc ggcgatgtgg gccccttcat 2281 catcggcttc ggcaattaca ggtatgccga ctacatgctg acctgcccaa tgctggtgta 2341 cgatctgctg tatcagctga gggcccccta tcgcgtgtct tgtagcgcca tcatctttgc 2401 catcctgatg tctggcgtgc tggccgagtt ctacgcagag ggcgacccta ggctgaggaa 2461 tggcgcctac gcctggtatg gctttggctg tttctggttt atcttcgcct actctatcgt 2521 gatgagcatc gtggccaagc agtatagccg gctggcccag ctggcccagg atacaggcgc 2581 cgagcacagc ctgcacgtgc tgaagttcgc cgtgtttacc ttctccatgc tgtggattct 2641 gtttcccctg gtgtgggcca tctgccctag aggcttcggc tggatcgacg ataactggac 2701 agaggtggcc cactgcgtgt gcgacatcgt ggccaagtcc tgttacggct ttgccctggc 2761 ccggttcaga aagacctatg atgaggagct gtttcggctg ctggagcagc tgggacacga 2821 cgaggatgag ttccagaagc tggagctgga tatgaggctg agcagcaatg gcgagcgcct 2881 gcggagactg tctgcggccg ccaagagcag gatcaccagc gagggcgagt acatccccct 2941 ggaccagatc gacatcaacg tgcagtccca gcccatcctc aacaccaagg agatggcccc 3001 gcagagcaag cctccagagg agctggagat gagcagcatg cccagccccg tggcccctct 3061 gcccgcacgc acggagggcg tcatcgacat gcggagcatg tccagcattg acagcttcat 3121 cagctgtgcc acggacttcc ctgaagccac cagattctac ccatacgatg ttccagatta 3181 cgcttaagaa ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga 3241 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt 3301 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa 3361 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt 3421 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct 3481 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg 3541 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc 3601 ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg 3661 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct 3721 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc 3781 cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat 3841 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc 3901 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat 3961 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg 4021 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca 4081 atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc 4141 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc 4201 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc 4261 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag 4321 gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct 4381 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct 4441 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc 4501 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata 4561 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac 4621 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc 4681 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt 4741 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg 4801 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag 4861 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt 4921 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt 4981 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat 5041 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc 5101 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag 5161 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt 5221 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg 5281 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa 5341 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa 5401 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct 5461 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg 5521 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg 5581 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt 5641 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga 5701 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga 5761 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac 5821 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg 5881 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac 5941 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct 6001 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct 6061 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg 6121 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat 6181 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg 6241 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat 6301 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct 6361 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa 6421 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa 6481 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc 6541 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta 6601 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct 6661 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg 6721 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag 6781 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc 6841 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg 6901 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt 6961 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg 7021 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca //