LOCUS Exported 6816 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION encodes pAAV NovArch. ACCESSION . VERSION . KEYWORDS MPC026: pAAV-CAG-FLEX-NovArch-TS-Citrine_TSx3-ER2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6816) 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 6816) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6816 /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(1520..2230) /codon_start=1 /product="enhanced variant of YFP (Heikal et al., 2001)" /label=Citrine /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELY" 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(2300..3058) /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 3055..3064 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind 3081..3114 /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 3185..3218 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 3260..3848 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3313..3333) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3731..3742) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3891..4012 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3902..3921) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3956..3975 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 4174..4303 /label=AAV2 ITR primer_bind complement(4369..4386) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4540..5128) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4620..4639) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5299..6159) /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 5922..5941 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6160..6264) /gene="bla" /label=AmpR promoter rep_origin complement(6290..6745) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6427..6448) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6639..6658 /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 agccccccac attgatgtca atctgatcga ggggtatata ctcgccctca gaagtgattc 1381 gactttttcc ccccacgttt atatcgattt gatccagagg gatgtactcg ccctcagagg 1441 ttatgcggga ttttccaccg acgttaatgt cgatctgatc cagagggata tactcgccct 1501 cagaggtgat ccttgatttg 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 tacatttatg tctatttgat ccagggggat gtactcgcct tcgcttgtga 2281 ttctactctt catcaacgta tcggcaccgg cactgggttc tggtgcctcg gtgtcgccca 2341 gaatagcccg ggatctcaac aggataaagc caaagccgac cttggcagtc acgtccaaca 2401 ccataaacag cagagtttcg gtgcccaggc ctatcacgcc agcgccctca gtgcctatga 2461 tccacaggat agggtaagcg gtccacagca ccaagaccag agctgtcagg gtgttaaagg 2521 tagatgccac ctcggggccc cgctcccttg cagcagatcg cagggatgta gccagaacat 2581 agagcaccac tatcatgcaa attgtagaga acaaccacca actgtatctg gctatggccg 2641 tgtggctcaa ggctccgatg aggccagtga cgatcatcag ggcgtccaca cccaccaggg 2701 tgccgatggt cacccgatcc accttagcga gaagggccag gtgcagcagc agaagtgggg 2761 tggtaaacag ccactgggcg tacctggcat aatagatatc caacatttcg cccccgacgg 2821 acacctcagt aagcccgata ccaaagaaca tagacagata tgcggcggat gcgattccgg 2881 aggccaggat agtcacagcg taatattccc gggcatcctt atcggtgact ccccatccgc 2941 ggaccagaaa gtagaaggtt ccaatcagca tcagcagagt gccgatgccc agccacaggg 3001 tttcaggtct gccgtcaccc agtaggtcgt aaccagcctg cagagcgata cttaccatgg 3061 tggcggtacc gaattctagt ataacttcgt ataggatact ttatacgaag ttatcattgg 3121 gattcttcct attttgatcc aagcatcacc atcgaccctc tagtccagat ctcaccatcg 3181 acccataact tcgtatagca tacattatac gaagttatgt ccctcgaaga ggttcgaatt 3241 cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg 3301 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta 3361 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct 3421 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt 3481 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac 3541 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg 3601 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc 3661 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg 3721 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct 3781 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc 3841 ctccccgcat cgataccgtc gacccgggcg gccgcttcga gcagacatga taagatacat 3901 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat 3961 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa 4021 caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcaa 4081 gtaaaacctc tacaaatgtg gtaaaatcga taaggatctt cctagagcat ggctacgtag 4141 ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg gagttggcca 4201 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc 4261 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcatt aatgaatcgg 4321 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 4381 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 4441 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 4501 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 4561 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 4621 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 4681 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc 4741 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 4801 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 4861 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 4921 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 4981 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 5041 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 5101 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 5161 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 5221 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 5281 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 5341 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 5401 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 5461 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 5521 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 5581 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 5641 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 5701 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 5761 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 5821 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 5881 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 5941 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 6001 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 6061 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 6121 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 6181 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 6241 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc 6301 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa 6361 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt 6421 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg 6481 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt 6541 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga 6601 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg 6661 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg 6721 cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa 6781 gggcgatcgg tgcgggcctc ttcgctatta cgccag //