LOCUS Exported 6046 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV-mediated expression of Archon1-KGC-GFP-ER2 under the Syn promoter. Using bGHpA signal.. ACCESSION . VERSION . KEYWORDS pAAV-Syn-Archon1-KGC-GFP-ER2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6046) 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 6046) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6046 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 349..796 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" regulatory 809..818 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1682..2395 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(1724..1745) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1985..2004) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2332..2353 /label=EGFP-C /note="EGFP, forward primer" misc_feature 2444..3032 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2497..2517) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2915..2926) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3050..3257 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 3309..3449 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3309..3438 /label=AAV2 ITR rep_origin 3524..3979 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3611..3630) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3821..3842 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3996..4015) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4115..4137 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4175..4193) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4261..4365 /gene="bla" /label=AmpR promoter CDS 4366..5226 /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(4584..4603) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5397..5985 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5886..5905 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct gcggccgcac gcgttgcaaa gatggataaa gttttaaaca gagaggaatc 181 tttgcagcta atggaccttc taggtcttga aaggagtggg aattggctcc ggtgcccgtc 241 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcagca 301 aattaattaa tggttaatta atctagactg cagagggccc tgcgtatgag tgcaagtggg 361 ttttaggacc aggatgaggc ggggtggggg tgcctacctg acgaccgacc ccgacccact 421 ggacaagcac ccaaccccca ttccccaaat tgcgcatccc ctatcagaga gggggagggg 481 aaacaggatg cggcgaggcg cgtgcgcact gccagcttca gcaccgcgga cagtgccttc 541 gcccccgcct ggcggcgcgc gccaccgccg cctcagcact gaaggcgcgc tgacgtcact 601 cgccggtccc ccgcaaactc cccttcccgg ccaccttggt cgcgtccgcg ccgccgccgg 661 cccagccgga ccgcaccacg cgaggcgcga gatagggggg cacgggcgcg accatctgcg 721 ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc 781 gtgcctgaga gcgcagtcga gaggatccgc caccatggtt tctatcgcac tgcaggctgg 841 ttatgacctc ctcggagatg gtagacctga aagtctgtgg ctcggaattg gtacactgct 901 gatgctgatc ggaaccttct acttcctggt tagagcgtgg ggagagactg acaaggatgc 961 acgggagtac tacgcagtga caatcctggt cagcggtatt gccagtgcag cttatctgtc 1021 aatgttcttt ggtattggcc tcacagaggt gccagtaggt ggagagatgc tgaacatcta 1081 ctacgcaaga tacgcacagt ggctgttcac tacaccactg ttgctgctgc acctggcact 1141 cctggccaaa gttgatagag tgaccatcgg cactctcgtt ggcgtcgatg ctctgatgat 1201 tgtcacaggc ctgataggaa cactgtcaca tactgccatt gccagatact cttggtggct 1261 gttctcaact atctgtatga tcgtcgttct gtatgtcctg gccacttctc tgcggtcagc 1321 cgctaaggag agaggcccag aggttgcctc aaccttcaac atcctcaccg cactggttct 1381 ggtcctgtgg actgcttacc ctatcatctg gattatcggc accgagggtg ccggagttgt 1441 gggcctggga atcgaaactt tgctgttcat ggtgctggac gtcaccgcca aggtgggctt 1501 cggttttatc ctgctcaggt caagagccat tctgcaagat acagaagcac ctgaaccttc 1561 agctggagct gataccggtg ccgccgaccg cccggtagta gcagtgagca aggcggccgc 1621 caagagcagg atcaccagcg agggcgagta catccccctg gaccagatcg acatcaacgt 1681 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg 1741 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg 1801 caagctgacc ctgaagttca tttgcaccac cggcaagctg cccgtgccct ggcccaccct 1861 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca 1921 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt 1981 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt 2041 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa 2101 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg 2161 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga 2221 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta 2281 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct 2341 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagttctg 2401 ctacgagaac gaggtgtaag aattccgtac aaagcttatc gataatcaac ctctggatta 2461 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg 2521 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc 2581 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca 2641 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac 2701 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact 2761 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc 2821 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctatg ttgccacctg 2881 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc 2941 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac 3001 gagtcggatc tccctttggg ccgcctcccc gcatcgatac cggctcgagc tgtgccttct 3061 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc 3121 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt 3181 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat 3241 agcaggcatg ctggggatgc ggtctcgagc tgattttgta ggtaaccacg tgcggaccga 3301 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct 3361 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt 3421 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca 3481 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc 3541 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 3601 ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 3661 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc 3721 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 3781 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 3841 ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt 3901 tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa 3961 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag 4021 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc 4081 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt 4141 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag 4201 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg 4261 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga 4321 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat 4381 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca 4441 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc 4501 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca 4561 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg 4621 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca 4681 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata 4741 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag 4801 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg 4861 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca 4921 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta 4981 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct 5041 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca 5101 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag 5161 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat 5221 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt 5281 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa 5341 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 5401 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 5461 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 5521 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag 5581 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 5641 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 5701 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 5761 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 5821 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 5881 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 5941 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 6001 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt //