LOCUS Exported 5833 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Genetically encoded voltage sensor ArcLight- codon optimized. ACCESSION . VERSION . KEYWORDS hSyn ArcLightCo (Q239)-T2A-nls-mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5833) AUTHORS Platisa J, Vasan G, Yang A, Pieribone VA TITLE Directed evolution of key residues in fluorescent protein inverses the polarity of voltage sensitivity in the genetically-encoded indicator ArcLight. JOURNAL ACS Chem Neurosci. 2017 Jan 3. doi: 10.1021/acschemneuro.6b00234. PUBMED 28045247 REFERENCE 2 (bases 1 to 5833) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5833 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 27..474 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" CDS 1963..2016 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 2038..2058 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2059..2766 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE GRHSTGGMDELYK" primer_bind complement(2201..2219) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 2469..2488 /label=mCherry-F /note="mCherry, forward primer" primer_bind 2663..2686 /label=DsRed1-C /note="DsRed1, forward primer" primer_bind complement(2856..2875) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 2921..2976 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(2975..2994) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(3335..3351) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3335..3351) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3348..3370) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3359..3375 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(3383..3413) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3428..3449 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 3508..3703 /label=SV40 promoter /note="SV40 early promoter" rep_origin 3554..3689 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3616..3635 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 3709..3843 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3746..3765) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3800..3819 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3911..3928) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4082..4670) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4162..4181) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4841..5701) /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 5464..5483 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5702..5806) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcgactgca gagggccctg cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg 61 ggtgggggtg cctacctgac gaccgacccc gacccactgg acaagcaccc aacccccatt 121 ccccaaattg cgcatcccct atcagagagg gggaggggaa acaggatgcg gcgaggcgcg 181 tgcgcactgc cagcttcagc accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc 241 caccgccgcc tcagcactga aggcgcgctg acgtcactcg ccggtccccc gcaaactccc 301 cttcccggcc accttggtcg cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg 361 aggcgcgaga taggggggca cgggcgcgac catctgcgct gcggcgccgg cgactcagcg 421 ctgcctcagt ctgcggtggg cagcggagga gtcgtgtcgt gcctgagagc gcagaccggt 481 accaccatgg aaggttttga cgggtcagac tttagccctc ctgccgatct cgtgggtgtg 541 gacggtgctg tgatgcggaa cgtggtggac gtgacaatca acggcgacgt gactgcacca 601 cctaaggcag ctcctagaaa atctgagagt gtgaagaaag tccactggaa cgacgtggat 661 cagggaccaa gcgagaagcc cgaaacaaga caggagaaga gcaggatcac cagcgagggc 721 gagtacatcc ccctggacca gatcgacatc aacgtggacg acgggaggat ggaaattcct 781 accacaggag tgggtagagt ccagtttagg gtgagagctg tcatcgacca tctgggaatg 841 cgggtgttcg gcgtctttct catcttcctg gacatcattc tgatgatcat tgatctgagt 901 ctccctggga agtcagagag ctcccagagc ttctacgacg gcatggcact ggccctcagc 961 tgctacttca tgctggatct cggcctgagg atcttcgcat acggacccaa gaacttcttt 1021 accaatcctt gggaggtggc cgacggactg atcattgtgg tcacttttgt ggtcaccatt 1081 ttctacacag tgctggacga gtacgtgcag gagacaggag cagacggact cggacagctg 1141 gtggtcctcg ctcgactgct ccgagtggtc cgactggcaa ggatctttta ctcccaccag 1201 caggggaatt caatgtctaa aggggaggaa ctgttcactg gtgtggtccc cattctggtg 1261 gagctcgacg gcgacgtgaa cggccataag ttttcagtga gcggggaggg tgaaggcgac 1321 gctacttacg gaaagctcac cctgaaattc atctgcacta ccgggaagct gccagtgcct 1381 tggcctaccc tggtcacaac tctcacatac ggcgtgcagt gtttcagtcg ctatcctgac 1441 catatgaagc gacacgattt ctttaaatct gctatgccag agggctacgt gcaggaacgg 1501 accatcttct ttaaggacga cggcaattat aaaactcgcg cagaggtgaa gtttgaaggc 1561 gacaccctgg tcaaccgaat cgagctcaag ggaattgact tcaaagaaga cggaaatatc 1621 ctggggcaca agctcgagta caactataac gaccatcagg tgtacatcat ggccgataag 1681 cagaaaaacg gcattaaggc taacttcaag atccgccaca acattgaaga cggcggagtg 1741 cagctggccg atcattacca gcagaacaca ccaatcggag acgggcctgt cctgctccca 1801 gataatcact atctgttcac cacatccact ctgtctaagg accccaacga gaaaagagat 1861 catatggtgc tgctggagtt tgtcactgcc gacgggatta cccacggaat ggacgagctg 1921 tataagttct gctacgagaa cgaggtggca tgcggaagcg gagaaggacg gggtagtctg 1981 ctgacatgcg gcgacgtaga agagaatcct ggtcccgccc cggggtcgac catggtgccc 2041 aagaagaaga ggaaagtcgt gagcaagggc gaggaggaca acatggccat catcaaggag 2101 ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag 2161 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag 2221 ggcggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag 2281 gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc 2341 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac 2401 tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc 2461 tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg 2521 taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc 2581 ggccactacg acgccgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc 2641 ggcgcctaca acgtcaacat caagctggac atcacctccc acaacgagga ctacaccatc 2701 gtggaacagt acgagcgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac 2761 aagtaagcta gcggccgcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt 2821 gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat 2881 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc 2941 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca 3001 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgccatatgc 3061 tggctgccat gaacaaaggt ggctataaag aggtcatcag tatatgaaac agccccctgc 3121 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg 3181 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag 3241 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatgaagatc 3301 cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt 3361 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg 3421 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt 3481 cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc 3541 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg 3601 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca 3661 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt 3721 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt 3781 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg 3841 atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 3901 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 3961 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 4021 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 4081 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 4141 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 4201 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 4261 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 4321 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 4381 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 4441 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 4501 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 4561 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 4621 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 4681 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 4741 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 4801 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 4861 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 4921 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 4981 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 5041 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 5101 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 5161 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 5221 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 5281 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 5341 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 5401 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 5461 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 5521 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 5581 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 5641 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 5701 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 5761 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 5821 aagtgccacc tgg //