LOCUS Exported 6518 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Codon-optimized Archon2 fluorescent voltage reporter in zebrafish expression plasmid. ACCESSION . VERSION . KEYWORDS pTol2-UAS-zArchon2-KGC-EGFP-ER2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6518) 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 6518) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6518 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 708..732 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(960..984) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" regulatory 992..1001 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1865..2578 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" polyA_signal 3908..3989 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3945..3964) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3999..4018 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4604..4621) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4775..5363) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4855..4874) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5534..6394) /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 6157..6176 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6395..6499) /gene="bla" /label=AmpR promoter ORIGIN 1 tgtccaggag ttcttgacag aggtgtaaaa agtactcaaa aattttactc aagtgaaagt 61 acaagtactt agggaaaatt ttactcaatt aaaagtaaaa gtatctggct agaatcttac 121 ttgagtaaaa gtaaaaaagt actccattaa aattgtactt gagtattaag gaagtaaaag 181 taaaagcaag aaagaaaact agagattctt gtttaagctt ttaatctcaa aaaacattaa 241 atgaaatgca tacaaggttt tatcctgctt tagaactgtt tgtatttaat tatcaaacta 301 taagacagac aatctaatgc cagtacacgc tactcaaagt tgtaaaacct cagatttaac 361 ttcagtagaa gctgattctc aaaattgtta gtgtcaagcc tagctctttt ggggctgaaa 421 agcaatcctg cagtgctgaa aagcctctca caggcagccg atgcgggaag aggtgtatta 481 gtcttgatag agaggctgca aatagcagga aacgtgagca gagactccct ggtgtctgaa 541 acacaggcca gatgggccat tcgtacgctt aatttcgaat atcgatatca agggcccatc 601 tcgagtatcc ggacacctag gtagcatgca tggcgccttg tcgactagct agcatcccgg 661 gaagaattcc tcgacggatc caccggtgtg gaattctgca gatatcaaca agtttgtaca 721 aaaaagcagg ctggccatca agcttaggcc tccacggtgg cttctaatcc gtgagtccta 781 gcgggtgaca gccctccgtc ttcacaggcg gaggagagtc ttccgtaggg ttcctcggag 841 tactgtcctc cgacgcgtgc aagggtcgac tctagagggt atataatgga tcccatcgcg 901 tctcagcctc actttgagct cctccacacg aattccctcg acctcgaaga cgcgtggata 961 cccagctttc ttgtacaaag tggtcactag tgccaccatg gtgagcatcg ctctgcaggc 1021 tggatacgac ctgctgggag acggaagacc tgagacactg tggctgggaa tcggaacact 1081 gctgatgctg atcggaacat tctacttcct ggtgagagga tggggagtga cagacaagga 1141 cgctagagag tactacgctg tgcctatcct ggtgagcgga atcgctagcg ctgcttacct 1201 gagcatgttc ttcggaatcg gactgacaga ggtgcctgtg ggaggagaga tgctggacat 1261 ctactacgct agatacgctc actggctgtt cagcacacct ctgctgctgc tggacctggc 1321 tctgctggct aaggtggaca gagtgatcat cggaacactg gtgggagtgg acgctctgat 1381 gatcgtgaca ggactgatcg gagctctgag ccacacagct atcgctagat acagctggtg 1441 gctgttcagc acaatctgta tgatcgtggt gctgtacgtg ctggctacaa gcctgagaag 1501 cgctgctaag gagagaggac ctgaggtggc tagcacattc aacatcctga cagctctggt 1561 gctggtgctg tggacagctt accctatcat ctggatcatc ggaacagagg gagctggagt 1621 ggtgggactg ggaatcgaga cactgctgtt catggtgctg gacgtgacat gtaaggtggg 1681 attcggattc atcctgctga gaagcagagc tatcctggga gacacagagg ctcctgagcc 1741 tagcgctgga gctgacagat ctgctgctga cagacctgtg gtggctgtga gcaaggctgc 1801 tgctaagagc agaatcacaa gcgagggaga gtacatccct ctggaccaga tcgacatcaa 1861 cgtggtgagc aagggagagg agctgttcac aggagtggtg cctatcctgg tggagctgga 1921 cggagacgtg aacggacaca agttcagcgt gagcggagag ggagagggag acgctacata 1981 cggaaagctg acactgaagt tcatctgtac aacaggaaag ctgcctgtgc cttggcctac 2041 actggtgaca acactgacat acggagtgca gtgtttcagc agataccctg accacatgaa 2101 gcagcacgac ttcttcaaga gcgctatgcc tgagggatac gtgcaggaga gaacaatctt 2161 cttcaaggac gacggaaact acaagacaag agctgaggtg aagttcgagg gagacacact 2221 ggtgaacaga atcgagctga agggaatcga cttcaaggag gacggaaaca tcctgggaca 2281 caagctggag tacaactaca acagccacaa cgtgtacatc atggctgaca agcagaagaa 2341 cggaatcaag gtgaacttca agatcagaca caacatcgag gacggaagcg tgcagctggc 2401 tgaccactac cagcagaaca cacctatcgg agacggacct gtgctgctgc ctgacaacca 2461 ctacctgagc acacagagcg ctctgagcaa ggaccctaac gagaagagag accacatggt 2521 gctgctggag ttcgtgacag ctgctggaat cacactggga atggacgagc tgtacaagtt 2581 ctgttacgag aacgaggtgt gaggcgcgcc aacccgctcc tcctccctcc ctccctccct 2641 ccctcctctc ctttattcac ttttcaacat gcacaacacg tcatcaaatc ccccaatcag 2701 cactgaatta cagccacggg ccaccagact gaaggtgtta aagaagacga ttcggtctca 2761 tcagcggcta aaacgagatg agggtgaacc gagagagact gtttccacag atgttcttga 2821 aggagggccg ggctcgtagg tgcggttatt acacatataa gctcccattc acaccttaaa 2881 acatccaaca gcattccaaa gttttccatc ggggaccagg gatagaaaac actgcctctt 2941 ccactgccta atttaacaca ttccttctag attatctctg gtcatctaga tgaaatcagt 3001 tctgttctgt ttgccgtgtc tgaaaagaca gtaaagagta attgaaaaaa aaaatgaata 3061 tagaaaaatg gggactggag agatatttgg actagacgct cctgatccag ccagaaggaa 3121 cctttcaggt gtcgattaca ctgcctgcga aaacaacctc ttcttccatt cttcagtttc 3181 cagctgccgg ctggcttctt ccactgccca acatcctaac aaacccctga caaaaaagga 3241 gacaagcctt gaaacatcct gaaataagca attcttagct gccagggacg tcttcctgtt 3301 agcatcagtg tatttgtttc ttttgctttg atttgttttc tttttttatt ttattttcat 3361 ggtaaatgta ttctgcctga gcactttaaa cgaaggtgtt tttgttccag tcgatgcttc 3421 gttttgaacc tgcttttata tttattaact tttgttgtgt atttttaatg tttaatcttt 3481 aaaggaatag ttcacctaca cacacacaca cacacacaca cacacacaca aatgaaatga 3541 agtcatcatt tactcacttt tctttctttt agctgtggag cacaaaacga gatgttttga 3601 agaatgttca cgctgttgtt tttcatgcaa tttaagtaaa cgaagatcaa gggatgttga 3661 gctatacaaa tattcattat atgggcagca cggtcgctgg ttcgagtccc ggctgggcca 3721 gttggcattt ctgagtttgc atattctccc tgtgttggca tgggtttcct cctggtgctc 3781 cgtttctccc acagtccact agatcataat cagccatacc acatttgtag aggttttact 3841 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt 3901 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa 3961 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa 4021 tgtatcaggc cttagggata acagggtaat acgcgtgatc tgcgaagata cggccacggg 4081 tgctcttgat cctgtggctg attttggact gtgctgctcg cagctgctga tgaatcacat 4141 acttcctcca ttttcttcca ctgattgact gttataattt ccctaatttc caggtcaagg 4201 tgctgtgcat tgtggtaata gatgtgacat gacgtcactt ccaaaggacc aatgaacatg 4261 tctgaccaat ttcatataat gtgaaaacga ttttcatagg cagaataaat aacatttaaa 4321 ttaaactggg catcagcgca attcaattgg tttggtaata gcaagggaaa atagaatgaa 4381 gtgatctcca aaaaataagt actttttgac tgtaaataaa attgtaagga gtaaaaagta 4441 cttttttttc taaaaaaatg taattaagta aaagtaaaag tattgatttt taattgtact 4501 caagtaaagt aaaaatcccc aaaaataata cttaagtaca gtaatcaagt aaaattactc 4561 aagtacttta cacctctggt tcttgaggta cctcttccgc ttcctcgctc actgactcgc 4621 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 4681 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 4741 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 4801 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 4861 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 4921 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 4981 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 5041 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 5101 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 5161 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag 5221 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 5281 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 5341 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 5401 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 5461 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 5521 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 5581 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct 5641 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 5701 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 5761 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 5821 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 5881 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 5941 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 6001 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 6061 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 6121 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 6181 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 6241 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 6301 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 6361 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 6421 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 6481 aacaaatagg ggttccgcgc acatttcccc gaggtacc //