LOCUS Exported 7204 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV expression of CaMKII-driven chimeric opsin variant C1V1 (E122T/E162T) for fast and potent optical excitation at red-shifted wavelengths.. ACCESSION . VERSION . KEYWORDS pAAV-CaMKIIa-C1V1 (t/t)-TS-EYFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7204) AUTHORS Yizhar O, Fenno LE, Prigge M, Schneider F, Davidson TJ, O'Shea DJ, Sohal VS, Goshen I, Finkelstein J, Paz JT, Stehfest K, Fudim R, Ramakrishnan C, Huguenard JR, Hegemann P, Deisseroth K TITLE Neocortical excitation/inhibition balance in information processing and social dysfunction. JOURNAL Nature. 2011 Jul 27;477(7363):171-8. doi: 10.1038/nature10360. PUBMED 21796121 REFERENCE 2 (bases 1 to 7204) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7204 /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 160..1448 /label=CaMKII promoter /note="promoter for the mouse calcium/calmodulin-dependent kinase II alpha subunit" CDS 2590..3306 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(2632..2653) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2893..2912) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3240..3261 /label=EGFP-C /note="EGFP, forward primer" misc_feature 3331..3919 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3384..3404) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3802..3813) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3951..4427 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4150..4169) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4467..4607 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4467..4596 /label=AAV2 ITR rep_origin 4682..5137 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4769..4788) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4979..5000 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5154..5173) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5273..5295 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5333..5351) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5419..5523 /gene="bla" /label=AmpR promoter CDS 5524..6384 /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(5742..5761) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6555..7143 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7044..7063 /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 cgcgtttaac attatggcct taggtcactt 181 catctccatg gggttcttct tctgattttc tagaaaatga gatgggggtg cagagagctt 241 cctcagtgac ctgcccaggg tcacatcaga aatgtcagag ctagaacttg aactcagatt 301 actaatctta aattccatgc cttgggggca tgcaagtacg atatacagaa ggagtgaact 361 cattagggca gatgaccaat gagtttagga aagaagagtc cagggcaggg tacatctaca 421 ccacccgccc agccctgggt gagtccagcc acgttcacct cattatagtt gcctctctcc 481 agtcctacct tgacgggaag cacaagcaga aactgggaca ggagccccag gagaccaaat 541 cttcatggtc cctctgggag gatgggtggg gagagctgtg gcagaggcct caggaggggc 601 cctgctgctc agtggtgaca gataggggtg agaaagcaga cagagtcatt ccgtcagcat 661 tctgggtctg tttggtactt cttctcacgc taaggtggcg gtgtgatatg cacaatggct 721 aaaaagcagg gagagctgga aagaaacaag gacagagaca gaggccaagt caaccagacc 781 aattcccaga ggaagcaaag aaaccattac agagactaca agggggaagg gaaggagaga 841 tgaattagct tcccctgtaa accttagaac ccagctgttg ccagggcaac ggggcaatac 901 ctgtctcttc agaggagatg aagttgccag ggtaactaca tcctgtcttt ctcaaggacc 961 atcccagaat gtggcaccca ctagccgtta ccatagcaac tgcctctttg ccccacttaa 1021 tcccatcccg tctgttaaaa gggccctata gttggaggtg ggggaggtag gaagagcgat 1081 gatcacttgt ggactaagtt tgttcgcatc cccttctcca accccctcag tacatcaccc 1141 tgggggaaca gggtccactt gctcctgggc ccacacagtc ctgcagtatt gtgtatataa 1201 ggccagggca aagaggagca ggttttaaag tgaaaggcag gcaggtgttg gggaggcagt 1261 taccggggca acgggaacag ggcgtttcgg aggtggttgc catggggacc tggatgctga 1321 cgaaggctcg cgaggctgtg agcagccaca gtgccctgct cagaagcccc aagctcgtca 1381 gtcaagccgg ttctccgttt gcactcagga gcacgggcag gcgagtggcc cctagttctg 1441 ggggcagctc tagagcggta ccggatcccc gggtaccggt aggccaccat gtcgcggagg 1501 ccatggcttc ttgccctagc gctggcagtg gcgctggcgg ccggcagcgc aggagcctcg 1561 actggcagtg acgcgacggt gccggtcgcg actcaggatg gccccgacta cgttttccac 1621 cgtgcccacg agcgcatgct cttccaaacc tcatacactc ttgagaacaa tggttctgtt 1681 atttgcatcc cgaacaacgg ccagtgcttc tgcttggctt ggcttaaatc caacggaaca 1741 aatgccgaga agttggctgc caacattctg cagtggatta cttttgcgct ttcagcgctc 1801 tgcctgatgt tctacggcta ccagacctgg aagtctactt gcggctggga gaccatttac 1861 gtggccacga tcgagatgat caagttcatc atcgagtatt tccatgagtt tgacgaacct 1921 gcggtgatct actcatccaa cggcaacaag accgtgtggc ttcgttacgc gacctggctg 1981 ctcacctgcc cggtccttct gatccatctg agcaacctca caggcctgaa ggacgattat 2041 agcaaaagga ctatgggcct gttggtttct gatgtgggat gcatcgtgtg gggcgcaacc 2101 agcgccatgt gtacggggtg gacgaagatc ctgttcttcc tcatctcatt gagctatggt 2161 atgtatacct attttcatgc tgctaaagtt tatatcgaag cattccacac agttccaaaa 2221 gggatttgtc gagaactggt ccgagtgatg gcctggacat tctttgtggc ttggggaatg 2281 tttccagtcc tgtttctgct gggcacggaa ggattcggtc atatcagccc ttatggatct 2341 gccattgggc actccatcct cgacctgatt gcaaagaaca tgtggggtgt gctggggaat 2401 tacctgcgcg tcaaaatcca cgagcacatc ctgttgtatg gcgacatcag aaagaagcag 2461 aaaattacga tcgccggcca agagatggag gttgagacac tggtggctga agaggaggac 2521 gcggccgcca agagcaggat caccagcgag ggcgagtaca tccccctgga ccagatcgac 2581 atcaacgtgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag 2641 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc 2701 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg 2761 cccaccctcg tgaccacctt cggctacggc ctgcagtgct tcgcccgcta ccccgaccac 2821 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc 2881 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac 2941 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg 3001 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag 3061 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag 3121 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac 3181 aaccactacc tgagctacca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac 3241 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac 3301 aagtaagaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa 3361 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta 3421 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa 3481 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg 3541 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc 3601 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc 3661 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg 3721 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg 3781 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg 3841 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc 3901 ctttgggccg cctccccgca tcgataccga gcgctgctcg agagatctac gggtggcatc 3961 cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca 4021 gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata 4081 ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg 4141 cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa 4201 tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc 4261 aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca 4321 tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca 4381 aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg attttgtagg 4441 taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc 4501 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt 4561 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg 4621 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag 4681 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc 4741 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc 4801 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt 4861 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg 4921 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt 4981 ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta 5041 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt 5101 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc 5161 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc 5221 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc 5281 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg 5341 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc 5401 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat 5461 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag 5521 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt 5581 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt 5641 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc 5701 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta 5761 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac 5821 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa 5881 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg 5941 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc 6001 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg 6061 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta 6121 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg 6181 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg 6241 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc 6301 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt 6361 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt 6421 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc 6481 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag 6541 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa 6601 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg 6661 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag 6721 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg 6781 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga 6841 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc 6901 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc 6961 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga 7021 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt 7081 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg 7141 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac 7201 atgt //