LOCUS Exported 7110 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Optogenetics. ACCESSION . VERSION . KEYWORDS pAAV-CaMKIIa-ChRmine-eYFP-WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7110) 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 7110) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7110 /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 154..1442 /label=CaMKII promoter /note="promoter for the mouse calcium/calmodulin-dependent kinase II alpha subunit" regulatory 1475..1484 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2477..3190 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(2519..2540) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2780..2799) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3127..3148 /label=EGFP-C /note="EGFP, forward primer" misc_feature 3242..3830 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3295..3315) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3713..3724) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3862..4338 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4061..4080) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4378..4518 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4378..4507 /label=AAV2 ITR rep_origin 4593..5048 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4680..4699) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4890..4911 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5065..5084) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5184..5206 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5244..5262) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5330..5434 /gene="bla" /label=AmpR promoter CDS 5435..6295 /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(5653..5672) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6466..7054 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6955..6974 /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 cgcacgcgtt taacattatg gccttaggtc acttcatctc 181 catggggttc ttcttctgat tttctagaaa atgagatggg ggtgcagaga gcttcctcag 241 tgacctgccc agggtcacat cagaaatgtc agagctagaa cttgaactca gattactaat 301 cttaaattcc atgccttggg ggcatgcaag tacgatatac agaaggagtg aactcattag 361 ggcagatgac caatgagttt aggaaagaag agtccagggc agggtacatc tacaccaccc 421 gcccagccct gggtgagtcc agccacgttc acctcattat agttgcctct ctccagtcct 481 accttgacgg gaagcacaag cagaaactgg gacaggagcc ccaggagacc aaatcttcat 541 ggtccctctg ggaggatggg tggggagagc tgtggcagag gcctcaggag gggccctgct 601 gctcagtggt gacagatagg ggtgagaaag cagacagagt cattccgtca gcattctggg 661 tctgtttggt acttcttctc acgctaaggt ggcggtgtga tatgcacaat ggctaaaaag 721 cagggagagc tggaaagaaa caaggacaga gacagaggcc aagtcaacca gaccaattcc 781 cagaggaagc aaagaaacca ttacagagac tacaaggggg aagggaagga gagatgaatt 841 agcttcccct gtaaacctta gaacccagct gttgccaggg caacggggca atacctgtct 901 cttcagagga gatgaagttg ccagggtaac tacatcctgt ctttctcaag gaccatccca 961 gaatgtggca cccactagcc gttaccatag caactgcctc tttgccccac ttaatcccat 1021 cccgtctgtt aaaagggccc tatagttgga ggtgggggag gtaggaagag cgatgatcac 1081 ttgtggacta agtttgttcg catccccttc tccaaccccc tcagtacatc accctggggg 1141 aacagggtcc acttgctcct gggcccacac agtcctgcag tattgtgtat ataaggccag 1201 ggcaaagagg agcaggtttt aaagtgaaag gcaggcaggt gttggggagg cagttaccgg 1261 ggcaacggga acagggcgtt tcggaggtgg ttgccatggg gacctggatg ctgacgaagg 1321 ctcgcgaggc tgtgagcagc cacagtgccc tgctcagaag ccccaagctc gtcagtcaag 1381 ccggttctcc gtttgcactc aggagcacgg gcaggcgagt ggcccctagt tctgggggca 1441 gctctagagc ggtaccggat ccccgggtac cggtgccacc atggcacacg caccaggcac 1501 cgaccagatg ttctacgtgg gcacaatgga cggctggtat ctggatacca agctgaactc 1561 cgtggccatc ggcgcccact ggtcttgctt tatcgtgctg acaatcacca cattctacct 1621 gggctatgag tcctggacat ctcggggccc tagcaagaga acctcctttt acgccggcta 1681 tcaggaggag cagaacctgg ccctgtttgt gaatttcttt gccatgctga gctacttcgg 1741 caagatcgtg gccgacacac tgggccacaa cttcggcgat gtgggcccct tcatcatcgg 1801 cttcggcaat tacaggtatg ccgactacat gctgacctgc ccaatgctgg tgtacgatct 1861 gctgtatcag ctgagggccc cctatcgcgt gtcttgtagc gccatcatct ttgccatcct 1921 gatgtctggc gtgctggccg agttctacgc agagggcgac cctaggctga ggaatggcgc 1981 ctacgcctgg tatggctttg gctgtttctg gtttatcttc gcctactcta tcgtgatgag 2041 catcgtggcc aagcagtata gccggctggc ccagctggcc caggatacag gcgccgagca 2101 cagcctgcac gtgctgaagt tcgccgtgtt taccttctcc atgctgtgga ttctgtttcc 2161 cctggtgtgg gccatctgcc ctagaggctt cggctggatc gacgataact ggacagaggt 2221 ggcccactgc gtgtgcgaca tcgtggccaa gtcctgttac ggctttgccc tggcccggtt 2281 cagaaagacc tatgatgagg agctgtttcg gctgctggag cagctgggac acgacgagga 2341 tgagttccag aagctggagc tggatatgag gctgagcagc aatggcgagc gcctgcggag 2401 actgtctgcg gccgccaaga gcaggatcac cagcgagggc gagtacatcc ccctggacca 2461 gatcgacatc aacgtggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct 2521 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg 2581 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt 2641 gccctggccc accctcgtga ccaccttcgg ctacggcctg cagtgcttcg cccgctaccc 2701 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga 2761 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga 2821 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa 2881 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga 2941 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag 3001 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct 3061 gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca acgagaagcg 3121 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga 3181 gctgtacaag ttctgctacg agaacgaggt gtaatgagaa ttcgatatca agcttatcga 3241 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 3301 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 3361 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 3421 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 3481 tggttgggga attgccacca cctgtcagct cctttccgga actttcgctt tccccctccc 3541 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 3601 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct 3661 cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 3721 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 3781 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg 3841 agcgctgctc gagagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg 3901 gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc 3961 attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc 4021 aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg 4081 agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc 4141 ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt 4201 ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa 4261 tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact 4321 gctcccttcc ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg 4381 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg 4441 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag 4501 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta 4561 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc 4621 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc 4681 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct 4741 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa 4801 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc 4861 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact 4921 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg 4981 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt 5041 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc 5101 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc 5161 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc 5221 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat 5281 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc 5341 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg 5401 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc 5461 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt 5521 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct 5581 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac 5641 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact 5701 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa 5761 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga 5821 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt 5881 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga 5941 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg 6001 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat 6061 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat 6121 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc 6181 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga 6241 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc 6301 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag 6361 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc 6421 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt 6481 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt 6541 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat 6601 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc 6661 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa 6721 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg 6781 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag 6841 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag 6901 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa 6961 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt 7021 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg 7081 gttcctggcc ttttgctggc cttttgctca //