LOCUS Exported 6753 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION encodes pAAV soma targeted NovArch. ACCESSION . VERSION . KEYWORDS MPC027: pAAV-CAG-FLEX-sNovArch-PP-Citrine_KV2.1TS-ER2 (soma ta... SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6753) AUTHORS Chien MP, Brinks D, Testa-Silva G, Tian H, Phil Brooks F 3rd, Adam Y, Bloxham W, Gmeiner B, Kheifets S, Cohen AE TITLE Photoactivated voltage imaging in tissue with an archaerhodopsin-derived reporter. JOURNAL Sci Adv. 2021 May 5;7(19). pii: 7/19/eabe3216. doi: 10.1126/sciadv.abe3216. Print 2021 May. PUBMED 33952514 REFERENCE 2 (bases 1 to 6753) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6753 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR promoter 441..718 /label=chicken beta-actin promoter primer_bind 997..1019 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1081..1100 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" protein_bind complement(1135..1168) /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(1231..1264) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS complement(1517..2230) /codon_start=1 /product="enhanced variant of YFP (Heikal et al., 2001)" /label=Citrine /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(1559..1580) /label=EGFP-C /note="EGFP, forward primer" primer_bind 1908..1927 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2167..2188 /label=EGFP-N /note="EGFP, reverse primer" CDS complement(2237..2995) /codon_start=1 /product="achaerhodopsin-3 (Arch) derivative with improved voltage sensitivity (Hochbaum et al., 2014)" /label=QuasAr2 /translation="MVSIALQAGYDLLGDGRPETLWLGIGTLLMLIGTFYFLVRGWGVT DKDAREYYAVTILASGIASAAYLSMFFGIGLTEVSVGGEMLDIYYARYAQWLFTTPLLL LHLALLAKVDRVTIGTLVGVDALMIVTGLIGALSHTAIARYSWWLFSTICMIVVLYVLA TSLRSAARERGPEVASTFNTLTALVLVLWTAYPILWIIGTEGAGVIGLGTETLLFMVLD VTAKVGFGFILLRSRAILGDTEAPEPSAGAD" regulatory 2992..3001 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind 3018..3051 /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 3122..3155 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 3197..3785 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3250..3270) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3668..3679) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3828..3949 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3839..3858) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3893..3912 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 4111..4240 /label=AAV2 ITR primer_bind complement(4306..4323) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4477..5065) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4557..4576) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5236..6096) /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 5859..5878 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6097..6201) /gene="bla" /label=AmpR promoter rep_origin complement(6227..6682) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6364..6385) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6576..6595 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct 181 aggaagagta ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac 241 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca 301 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg 361 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt 421 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc 481 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat 541 gggggcgggg gggggggggg ggcgcgcgcc gggggggggg gggggggggg gggggggggg 601 gggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc 661 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg 721 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc 781 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc 841 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag 901 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggc tgtccgcggg 961 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg 1021 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg 1081 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattggat ccccataact 1141 tcgtataaag tatcctatac gaagttatat caaaatagga agaccaatgc ttcaccatcg 1201 acccgaattg ccaagcatca ccatcgaccc ataacttcgt ataatgtatg ctatacgaag 1261 ttatactagg gatcccccct cgagttttac ttgtacatta cacttcattc tcataacaaa 1321 agaatctggt ggcttcaggg aagtccgtgg cacagctgat gaagctgtca atgctggaca 1381 tgctccgcat gtcgatgacg ccctccgtgc gtgcgggcag aggggccacg gggctgggca 1441 tgctgctcat ctccagctcc tctggaggct tgctctgcgg ggccatctcc ttggtgttga 1501 ggatgggctg ggactgcttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca 1561 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc agggcggact 1621 ggtagctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct 1681 gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt 1741 tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt 1801 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct 1861 cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc 1921 cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga 1981 agaagtcgtg ctgcttcatg tggtcggggt agcgggcgaa gcacatcagg ccgtagccga 2041 aggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca 2101 gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc 2161 cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc 2221 ccttgctcac cggaggatcg gcaccggcac tgggttctgg tgcctcggtg tcgcccagaa 2281 tagcccggga tctcaacagg ataaagccaa agccgacctt ggcagtcacg tccaacacca 2341 taaacagcag agtttcggtg cccaggccta tcacgccagc gccctcagtg cctatgatcc 2401 acaggatagg gtaagcggtc cacagcacca agaccagagc tgtcagggtg ttaaaggtag 2461 atgccacctc ggggccccgc tcccttgcag cagatcgcag ggatgtagcc agaacataga 2521 gcaccactat catgcaaatt gtagagaaca accaccaact gtatctggct atggccgtgt 2581 ggctcaaggc tccgatgagg ccagtgacga tcatcagggc gtccacaccc accagggtgc 2641 cgatggtcac ccgatccacc ttagcgagaa gggccaggtg cagcagcaga agtggggtgg 2701 taaacagcca ctgggcgtac ctggcataat agatatccaa catttcgccc ccgacggaca 2761 cctcagtaag cccgatacca aagaacatag acagatatgc ggcggatgcg attccggagg 2821 ccaggatagt cacagcgtaa tattcccggg catccttatc ggtgactccc catccgcgga 2881 ccagaaagta gaaggttcca atcagcatca gcagagtgcc gatgcccagc cacagggttt 2941 caggtctgcc gtcacccagt aggtcgtaac cagcctgcag agcgatactt accatggtgg 3001 cggtaccgaa ttctagtata acttcgtata ggatacttta tacgaagtta tcattgggat 3061 tcttcctatt ttgatccaag catcaccatc gaccctctag tccagatctc accatcgacc 3121 cataacttcg tatagcatac attatacgaa gttatgtccc tcgaagaggt tcgaattcga 3181 tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat 3241 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca 3301 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc 3361 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc 3421 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt 3481 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg 3541 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc 3601 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta 3661 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg 3721 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc 3781 cccgcatcga taccgtcgac ccgggcggcc gcttcgagca gacatgataa gatacattga 3841 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg 3901 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa 3961 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagcaagta 4021 aaacctctac aaatgtggta aaatcgataa ggatcttcct agagcatggc tacgtagata 4081 agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc 4141 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg 4201 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcattaat gaatcggcca 4261 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc 4321 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 4381 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 4441 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 4501 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 4561 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 4621 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 4681 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 4741 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 4801 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 4861 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac 4921 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 4981 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat 5041 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc 5101 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt 5161 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta 5221 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct 5281 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg 5341 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga 5401 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt 5461 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt 5521 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt 5581 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat 5641 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc 5701 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc 5761 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat 5821 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag 5881 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt 5941 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc 6001 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa 6061 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg 6121 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa 6181 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt 6241 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag 6301 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt 6361 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga 6421 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg 6481 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct 6541 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc 6601 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt 6661 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg 6721 cgatcggtgc gggcctcttc gctattacgc cag //