LOCUS Exported 6672 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION AAV production plasmid encoding for Archon1 fluorescent voltage reporter. ACCESSION . VERSION . KEYWORDS pAAV-CaMKII-Archon1-KGC-EGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6672) AUTHORS Piatkevich KD, Jung EE, Straub C, Linghu C, Park D, Suk HJ, Hochbaum DR, Goodwin D, Pnevmatikakis E, Pak N, Kawashima T, Yang CT, Rhoades JL, Shemesh O, Asano S, Yoon YG, Freifeld L, Saulnier JL, Riegler C, Engert F, Hughes T, Drobizhev M, Szabo B, Ahrens MB, Flavell SW, Sabatini BL, Boyden ES TITLE A robotic multidimensional directed evolution approach applied to fluorescent voltage reporters. JOURNAL Nat Chem Biol. 2018 Feb 26. pii: 10.1038/s41589-018-0004-9. doi: 10.1038/s41589-018-0004-9. PUBMED 29483642 REFERENCE 2 (bases 1 to 6672) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6672 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 35..164 /label=AAV2 ITR promoter 237..1525 /label=CaMKII promoter /note="promoter for the mouse calcium/calmodulin-dependent kinase II alpha subunit" regulatory 1536..1545 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2409..3122 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(2451..2472) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2712..2731) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3059..3080 /label=EGFP-C /note="EGFP, forward primer" misc_feature 3150..3738 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3203..3223) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3621..3632) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3781..3902 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3792..3811) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3846..3865 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 4064..4193 /label=AAV2 ITR primer_bind complement(4259..4276) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4430..5018) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4510..4529) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5189..6049) /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 5812..5831 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6050..6154) /gene="bla" /label=AmpR promoter rep_origin complement(6180..6635) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6317..6338) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6529..6548 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 gcgatcggtg cgggcctctt cgctattacg ccagctgcgc gctcgctcgc tcactgaggc 61 cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg 121 agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt taatgattaa 181 cccgccatgc tacttatcta cgtagccatg ctctaggaag atcgtaggtt aattaacatt 241 atggccttag gtcacttcat ctccatgggg ttcttcttct gattttctag aaaatgagat 301 gggggtgcag agagcttcct cagtgacctg cccagggtca catcagaaat gtcagagcta 361 gaacttgaac tcagattact aatcttaaat tccatgcctt gggggcatgc aagtacgata 421 tacagaagga gtgaactcat tagggcagat gaccaatgag tttaggaaag aagagtccag 481 ggcagggtac atctacacca cccgcccagc cctgggtgag tccagccacg ttcacctcat 541 tatagttgcc tctctccagt cctaccttga cgggaagcac aagcagaaac tgggacagga 601 gccccaggag accaaatctt catggtccct ctgggaggat gggtggggag agctgtggca 661 gaggcctcag gaggggccct gctgctcagt ggtgacagat aggggtgaga aagcagacag 721 agtcattccg tcagcattct gggtctgttt ggtacttctt ctcacgctaa ggtggcggtg 781 tgatatgcac aatggctaaa aagcagggag agctggaaag aaacaaggac agagacagag 841 gccaagtcaa ccagaccaat tcccagagga agcaaagaaa ccattacaga gactacaagg 901 gggaagggaa ggagagatga attagcttcc cctgtaaacc ttagaaccca gctgttgcca 961 gggcaacggg gcaatacctg tctcttcaga ggagatgaag ttgccagggt aactacatcc 1021 tgtctttctc aaggaccatc ccagaatgtg gcacccacta gccgttacca tagcaactgc 1081 ctctttgccc cacttaatcc catcccgtct gttaaaaggg ccctatagtt ggaggtgggg 1141 gaggtaggaa gagcgatgat cacttgtgga ctaagtttgt tcgcatcccc ttctccaacc 1201 ccctcagtac atcaccctgg gggaacaggg tccacttgct cctgggccca cacagtcctg 1261 cagtattgtg tatataaggc cagggcaaag aggagcaggt tttaaagtga aaggcaggca 1321 ggtgttgggg aggcagttac cggggcaacg ggaacagggc gtttcggagg tggttgccat 1381 ggggacctgg atgctgacga aggctcgcga ggctgtgagc agccacagtg ccctgctcag 1441 aagccccaag ctcgtcagtc aagccggttc tccgtttgca ctcaggagca cgggcaggcg 1501 agtggcccct agttctgggg gcagcggggg atcccgccac catggtttct atcgcactgc 1561 aggctggtta tgacctcctc ggagatggta gacctgaaag tctgtggctc ggaattggta 1621 cactgctgat gctgatcgga accttctact tcctggttag agcgtgggga gagactgaca 1681 aggatgcacg ggagtactac gcagtgacaa tcctggtcag cggtattgcc agtgcagctt 1741 atctgtcaat gttctttggt attggcctca cagaggtgcc agtaggtgga gagatgctga 1801 acatctacta cgcaagatac gcacagtggc tgttcactac accactgttg ctgctgcacc 1861 tggcactcct ggccaaagtt gatagagtga ccatcggcac tctcgttggc gtcgatgctc 1921 tgatgattgt cacaggcctg ataggaacac tgtcacatac tgccattgcc agatactctt 1981 ggtggctgtt ctcaactatc tgtatgatcg tcgttctgta tgtcctggcc acttctctgc 2041 ggtcagccgc taaggagaga ggcccagagg ttgcctcaac cttcaacatc ctcaccgcac 2101 tggttctggt cctgtggact gcttacccta tcatctggat tatcggcacc gagggtgccg 2161 gagttgtggg cctgggaatc gaaactttgc tgttcatggt gctggacgtc accgccaagg 2221 tgggcttcgg ttttatcctg ctcaggtcaa gagccattct gcaagataca gaagcacctg 2281 aaccttcagc tggagctgat accggtgccg ccgaccgccc ggtagtagca gtgagcaagg 2341 cggccgccaa gagcaggatc accagcgagg gcgagtacat ccccctggac cagatcgaca 2401 tcaacgtggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc 2461 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca 2521 cctacggcaa gctgaccctg aagttcattt gcaccaccgg caagctgccc gtgccctggc 2581 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca 2641 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca 2701 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca 2761 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg 2821 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga 2881 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc 2941 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca 3001 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca 3061 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca 3121 agtaagaatt cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa 3181 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa 3241 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat 3301 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt 3361 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc 3421 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc 3481 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg 3541 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga 3601 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc 3661 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc 3721 tttgggccgc ctccccgcat cgataccgtc gacccgggcg gccgcttcga gcagacatga 3781 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta 3841 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag 3901 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt 3961 tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatctt cctagagcat 4021 ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg 4081 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc 4141 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcatt 4201 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 4261 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 4321 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 4381 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 4441 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 4501 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 4561 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 4621 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 4681 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 4741 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 4801 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 4861 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 4921 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 4981 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 5041 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 5101 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 5161 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 5221 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 5281 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 5341 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 5401 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 5461 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 5521 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 5581 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 5641 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 5701 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 5761 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 5821 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 5881 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 5941 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 6001 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 6061 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 6121 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta 6181 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt 6241 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat 6301 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa 6361 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta 6421 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc 6481 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc 6541 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac 6601 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa 6661 ctgttgggaa gg //