LOCUS Exported 6579 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV-mediated expression of Archon1-KGC-GFP-ER2 under the EF1-alpha promoter (1.1kb short version). Using SV40 signal.. ACCESSION . VERSION . KEYWORDS pAAV-EF1-alpha-1.1-Archon1-KGC-GFP-ER2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6579) 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 6579) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6579 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(11..28) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(182..770) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(262..281) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(941..1801) /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 1564..1583 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1802..1906) /gene="bla" /label=AmpR promoter rep_origin complement(1932..2387) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2069..2090) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2281..2300 /label=F1ori-R /note="F1 origin, reverse primer" repeat_region 2459..2588 /label=AAV2 ITR promoter 2661..3839 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 2892..3830 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3787..3807 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" regulatory 3846..3855 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4719..5432 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(4761..4782) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(5022..5041) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5369..5390 /label=EGFP-C /note="EGFP, forward primer" misc_feature 5481..6069 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5534..5554) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5952..5963) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 6112..6233 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6123..6142) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 6177..6196 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 6395..6524 /label=AAV2 ITR ORIGIN 1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta 1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga 2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa 2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga 2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc 2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga 2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg 2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt 2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct 2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg 2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag 2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag 2641 gaagatcgta ggttaattaa ggctccggtg cccgtcagtg ggcagagcgc acatcgccca 2701 cagtccccga gaagttgggg ggaggggtcg gcaattgaac cggtgcctag agaaggtggc 2761 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg 2821 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg 2881 ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg 2941 gcccttgcgt gccttgaatt acttccacct ggctgcagta cgtgattctt gatcccgagc 3001 ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc 3061 gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc 3121 ttcgcgcctg tctcgctgct ttcgataagt ctctagccat ttaaaatttt tgatgacctg 3181 ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc gggccaagat ctgcacactg 3241 gtatttcggt ttttggggcc gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt 3301 cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg acgggggtag tctcaagctg 3361 gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa 3421 ggctggcccg gtcggcacca gttgcgtgag cggaaagatg gccgcttccc ggccctgctg 3481 cagggagctc aaaatggagg acgcggcgct cgggagagcg ggcgggtgag tcacccacac 3541 aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg tgactccacg gagtaccggg 3601 cgccgtccag gcacctcgat tagttctcga gcttttggag tacgtcgtct ttaggttggg 3661 gggaggggtt ttatgcgatg gagtttcccc acactgagtg ggtggagact gaagttaggc 3721 cagcttggca cttgatgtaa ttctccttgg aatttgccct ttttgagttt ggatcttggt 3781 tcattctcaa gcctcagaca gtggttcaaa gtttttttct tccatttcag gtgtcgtgag 3841 gatccgccac catggtttct atcgcactgc aggctggtta tgacctcctc ggagatggta 3901 gacctgaaag tctgtggctc ggaattggta cactgctgat gctgatcgga accttctact 3961 tcctggttag agcgtgggga gagactgaca aggatgcacg ggagtactac gcagtgacaa 4021 tcctggtcag cggtattgcc agtgcagctt atctgtcaat gttctttggt attggcctca 4081 cagaggtgcc agtaggtgga gagatgctga acatctacta cgcaagatac gcacagtggc 4141 tgttcactac accactgttg ctgctgcacc tggcactcct ggccaaagtt gatagagtga 4201 ccatcggcac tctcgttggc gtcgatgctc tgatgattgt cacaggcctg ataggaacac 4261 tgtcacatac tgccattgcc agatactctt ggtggctgtt ctcaactatc tgtatgatcg 4321 tcgttctgta tgtcctggcc acttctctgc ggtcagccgc taaggagaga ggcccagagg 4381 ttgcctcaac cttcaacatc ctcaccgcac tggttctggt cctgtggact gcttacccta 4441 tcatctggat tatcggcacc gagggtgccg gagttgtggg cctgggaatc gaaactttgc 4501 tgttcatggt gctggacgtc accgccaagg tgggcttcgg ttttatcctg ctcaggtcaa 4561 gagccattct gcaagataca gaagcacctg aaccttcagc tggagctgat accggtgccg 4621 ccgaccgccc ggtagtagca gtgagcaagg cggccgccaa gagcaggatc accagcgagg 4681 gcgagtacat ccccctggac cagatcgaca tcaacgtggt gagcaagggc gaggagctgt 4741 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca 4801 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcattt 4861 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg 4921 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca 4981 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga 5041 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca 5101 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc 5161 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc 5221 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca 5281 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga 5341 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg 5401 ggatcactct cggcatggac gagctgtaca agttctgcta cgagaacgag gtgtaagaat 5461 tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg 5521 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt 5581 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc 5641 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt 5701 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga 5761 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct 5821 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat 5881 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct 5941 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc 6001 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg 6061 cctccccgca tcgataccgt cgacccgggc ggccgcttcg agcagacatg ataagataca 6121 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa 6181 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca 6241 acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagca 6301 agtaaaacct ctacaaatgt ggtaaaatcg ataaggatct tcctagagca tggctacgta 6361 gataagtagc atggcgggtt aatcattaac tacaaggaac ccctagtgat ggagttggcc 6421 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc 6481 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcat taatgaatcg 6541 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgct //