LOCUS Exported 7412 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV expression of Ef1a-driven, cre-dependent, chimeric opsin variant C1V1 (E122T/E162T) for fast and potent optical excitation at red-shifted wavelengths.. ACCESSION . VERSION . KEYWORDS pAAV-Ef1a-DIO C1V1 (t/t)-TS-EYFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7412) 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 7412) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7412 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 81..210 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 311..1489 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 542..1480 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1437..1457 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" protein_bind 1520..1553 /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind 1604..1637 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS complement(1649..2365) /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(1694..1715) /label=EGFP-C /note="EGFP, forward primer" primer_bind 2043..2062 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2302..2323 /label=EGFP-N /note="EGFP, reverse primer" protein_bind complement(3477..3510) /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind complement(3561..3594) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 3619..4207 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3672..3692) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4090..4101) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4239..4715 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4438..4457) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4755..4895 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4755..4884 /label=AAV2 ITR rep_origin 4970..5425 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5057..5076) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5267..5288 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5442..5461) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5561..5583 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5621..5639) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5707..5811 /gene="bla" /label=AmpR promoter CDS 5812..6672 /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(6030..6049) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6843..19 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7332..7351 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg 61 ctggcctttt gctcacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc 121 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg 181 gagtggccaa ctccatcact aggggttcct gcggccgcac gcgtaagctt tgcaaagatg 241 gataaagttt taaacagaga ggaatctttg cagctaatgg accttctagg tcttgaaagg 301 agtgggaatt ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga 361 gaagttgggg ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa 421 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta 481 tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca 541 ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt 601 gccttgaatt acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg 661 aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt 721 tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg 781 tctcgctgct ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct 841 ttttttctgg caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt 901 ttttggggcc gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg 961 gggcctgcga gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc 1021 tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg 1081 gtcggcacca gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc 1141 aaaatggagg acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag 1201 ggcctttccg tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag 1261 gcacctcgat tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt 1321 ttatgcgatg gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca 1381 cttgatgtaa ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa 1441 gcctcagaca gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc 1501 ctctagagtc gactccggaa taacttcgta taggatactt tatacgaagt tatgcagaat 1561 ggtagctgga ttgtagctgc tattagcaat atgaaacctc ttaataactt cgtatagcat 1621 acattatacg aagttatggc gcgccctatt acttgtacag ctcgtccatg ccgagagtga 1681 tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt 1741 tgctcagggc ggactggtag ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc 1801 cgatgggggt gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt 1861 ggcggatctt gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt 1921 tgtggctgtt gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt 1981 cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc 2041 gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg 2101 gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcact 2161 gcaggccgta gccgaaggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg 2221 tgcagatgaa cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca 2281 cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg 2341 tgaacagctc ctcgcccttg ctcaccacgt tgatgtcgat ctggtccagg gggatgtact 2401 cgccctcgct ggtgatcctg ctcttggcgg ccgcgtcctc ctcttcagcc accagtgtct 2461 caacctccat ctcttggccg gcgatcgtaa ttttctgctt ctttctgatg tcgccataca 2521 acaggatgtg ctcgtggatt ttgacgcgca ggtaattccc cagcacaccc cacatgttct 2581 ttgcaatcag gtcgaggatg gagtgcccaa tggcagatcc ataagggctg atatgaccga 2641 atccttccgt gcccagcaga aacaggactg gaaacattcc ccaagccaca aagaatgtcc 2701 aggccatcac tcggaccagt tctcgacaaa tcccttttgg aactgtgtgg aatgcttcga 2761 tataaacttt agcagcatga aaataggtat acataccata gctcaatgag atgaggaaga 2821 acaggatctt cgtccacccc gtacacatgg cgctggttgc gccccacacg atgcatccca 2881 catcagaaac caacaggccc atagtccttt tgctataatc gtccttcagg cctgtgaggt 2941 tgctcagatg gatcagaagg accgggcagg tgagcagcca ggtcgcgtaa cgaagccaca 3001 cggtcttgtt gccgttggat gagtagatca ccgcaggttc gtcaaactca tggaaatact 3061 cgatgatgaa cttgatcatc tcgatcgtgg ccacgtaaat ggtctcccag ccgcaagtag 3121 acttccaggt ctggtagccg tagaacatca ggcagagcgc tgaaagcgca aaagtaatcc 3181 actgcagaat gttggcagcc aacttctcgg catttgttcc gttggattta agccaagcca 3241 agcagaagca ctggccgttg ttcgggatgc aaataacaga accattgttc tcaagagtgt 3301 atgaggtttg gaagagcatg cgctcgtggg cacggtggaa aacgtagtcg gggccatcct 3361 gagtcgcgac cggcaccgtc gcgtcactgc cagtcgaggc tcctgcgctg ccggccgcca 3421 gcgccactgc cagcgctagg gcaagaagcc atggcctccg cgacatggtg gctagcataa 3481 cttcgtataa agtatcctat acgaagttat ttgccttaac ccagaaatta tcactgttat 3541 tctttagaat ggtgcaaaga ataacttcgt ataatgtatg ctatacgaag ttatgaattc 3601 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt 3661 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat 3721 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg 3781 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt 3841 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact 3901 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc 3961 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg 4021 tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc 4081 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg 4141 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc 4201 tccccgcatc gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct 4261 ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa 4321 taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga 4381 ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct 4441 attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct 4501 gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga 4561 ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc 4621 tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat 4681 tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg 4741 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg 4801 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc 4861 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct 4921 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt 4981 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc 5041 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc 5101 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg 5161 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga 5221 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc 5281 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg 5341 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt 5401 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc 5461 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt 5521 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag 5581 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt 5641 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga 5701 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc 5761 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt 5821 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct 5881 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt 5941 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt 6001 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac 6061 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac 6121 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct 6181 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg 6241 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg 6301 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca 6361 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa 6421 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt 6481 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc 6541 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg 6601 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt 6661 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt 6721 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc 6781 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct 6841 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 6901 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc 6961 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac 7021 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct 7081 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat 7141 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg 7201 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa 7261 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg 7321 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga 7381 cttgagcgtc gatttttgtg atgctcgtca gg //