LOCUS       sequence-170997-        5835 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  genetically-encoded fluorescent voltage indicator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5835)
  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 5835)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Chem 
            Neurosci. 2017 Jan 3. doi: 10.1021/acschemneuro.6b00234."
COMMENT     Sequence Label: hSyn Marina-T2A-nls-mCherry
FEATURES             Location/Qualifiers
     source          1..5835
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(5..452)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     promoter        506..610
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(611..679,680..1471)
                     /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(829..848)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1642..2230
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2131..2150
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2384..2401
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    2469..2603
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2493..2512)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2547..2566
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(2609..2804)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2623..2758
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    2863..2884
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2899..2904,2905..2922,2923..2929)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2937..2953
                     /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)."
     primer_bind     2942..2964
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2961..2977
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2961..2977
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3320..3339
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    3338..3393
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     3439..3458
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(3548..4255)
                     /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(3628..3651)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(3826..3845)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4095..4113
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(4256..4276)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(4298..4351)
                     /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"
ORIGIN
        1 cggtctgcgc tctcaggcac gacacgactc ctccgctgcc caccgcagac tgaggcagcg
       61 ctgagtcgcc ggcgccgcag cgcagatggt cgcgcccgtg cccccctatc tcgcgcctcg
      121 cgtggtgcgg tccggctggg ccggcggcgg cgcggacgcg accaaggtgg ccgggaaggg
      181 gagtttgcgg gggaccggcg agtgacgtca gcgcgccttc agtgctgagg cggcggtggc
      241 gcgcgccgcc aggcgggggc gaaggcactg tccgcggtgc tgaagctggc agtgcgcacg
      301 cgcctcgccg catcctgttt cccctccccc tctctgatag gggatgcgca atttggggaa
      361 tgggggttgg gtgcttgtcc agtgggtcgg ggtcggtcgt caggtaggca cccccacccc
      421 gcctcatcct ggtcctaaaa cccacttgca ctcatacgca gggccctctg cagtcgaccc
      481 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
      541 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      601 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
      661 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
      721 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
      781 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
      841 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
      901 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
      961 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     1021 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     1081 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     1141 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     1201 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     1261 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     1321 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     1381 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     1441 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     1501 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     1561 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1621 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1681 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1741 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     1801 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1861 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1921 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1981 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     2041 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     2101 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     2161 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     2221 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     2281 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     2341 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     2401 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     2461 atgcagcgga tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     2521 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     2581 ttataagctg caataaacaa gttagctttt tgcaaaagcc taggcctcca aaaaagcctc
     2641 ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa
     2701 aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg
     2761 gcggagttag gggcgggact atggttgctg actaattgag atgcggatcc gctggcacga
     2821 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     2881 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     2941 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcttgggct
     3001 gcaggtcgag ggatctccat aagagaagag ggacagctat gactgggagt agtcaggaga
     3061 ggaggaaaaa tctggctagt aaaacatgta aggaaaattt tagggatgtt aaagaaaaaa
     3121 ataacacaaa acaaaatata aaaaaaatct aacctcaagt caaggctttt ctatggaata
     3181 aggaatggac agcagggggc tgtttcatat actgatgacc tctttatagc caacctttgt
     3241 tcatggcagc cagcatatgg gcatatgttg ccaaactcta aaccaaatac tcattctgat
     3301 gttttaaatg atttgccctc ccatatgtcc ttccgagtga gagacacaaa aaattccaac
     3361 acactattgc aatgaaaata aatttccttt attagccaga agtcagatgc tcaaggggct
     3421 tcatgatgtc cccataattt ttggcagagg gaaaaagatc tcagtggtat ttgtgagcca
     3481 gggcattggc cacaccagcc accaccttct gataggcagc ctgcacctga ggagtgcggc
     3541 cgctagctta cttgtacagc tcgtccatgc cgccggtgga gtggcggccc tcggcgcgct
     3601 cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt gatgtccagc ttgatgttga
     3661 cgttgtaggc gccgggcagc tgcacgggct tcttggcctt gtaggtggtc ttgacctcgg
     3721 cgtcgtagtg gccgccgtcc ttcagcttca gcctctgctt gatctcgccc ttcagggcgc
     3781 cgtcctcggg gtacatccgc tcggaggagg cctcccagcc catggtcttc ttctgcatta
     3841 cggggccgtc ggaggggaag ttggtgccgc gcagcttcac cttgtagatg aactcgccgt
     3901 cctgcaggga ggagtcctgg gtcacggtca ccacgccgcc gtcctcgaag ttcatcacgc
     3961 gctcccactt gaagccctcg gggaaggaca gcttcaagta gtcggggatg tcggcggggt
     4021 gcttcacgta ggccttggag ccgtacatga actgagggga caggatgtcc caggcgaagg
     4081 gcagggggcc gcccttggtc accttcagct tggcggtctg ggtgccctcg taggggcggc
     4141 cctcgccctc gccctcgatc tcgaactcgt ggccgttcac ggagccctcc atgtgcacct
     4201 tgaagcgcat gaactccttg atgatggcca tgttgtcctc ctcgcccttg ctcacgactt
     4261 tcctcttctt cttgggcacc atggtcgacc ccggggcggg accaggattc tcttctacgt
     4321 cgccgcatgt cagcagacta ccccgtcctt ctccgcttcc gcatgccacc tcgttctcgt
     4381 agcagaactt atacagctcg tccattccgt gggtaatccc gtcggcagtg acaaactcca
     4441 gcagcaccat atgatctctt ttctcgttgg ggtccttaga cagagtggat gtggtgaaca
     4501 gaacgtgatt atctgggagc aggacaggcc cgtctccgat tggtgtgttc tgctggtaat
     4561 gatcggccag ctgcactccg ccgtcttcaa tgttgtggcg gatcttgaag ttagccttaa
     4621 tgccgttttt ctgcttatcg gccatgatgt acacctgagc ggcgttatag ttgtactcga
     4681 gcttgtgccc caggatattt ccgtcttctt tgaagtcaat tcccttgagc tcgattcggt
     4741 tgaccagggt gtcgccttca aacttcacct ctgcgcgagt tttataattg ccgtcgtcct
     4801 taaagaagat ggtccgttcc tgcacgtagc cctctggcat agcagattta aagaaatcgt
     4861 gtcgcttcat atggtcagga tagcgactga aacactgcac gccgtatgtg agagttgtga
     4921 ccagggtagg ccaaggcact ggcagcttcc cggtagtgca gatgaatttc agggtgagct
     4981 ttccgtaagt agcgtcgcct tcaccctccc cgctcactga aaacttatgg ccgttcacgt
     5041 cgccgtcgag ctccaccaga atggggacca caccagtgaa cagttcctcc cctttagaca
     5101 ttgaattccc ctgctggtgg gagtaaaaga tccttgccag tcggaccact cggagcagtc
     5161 gagcgaggac caccagctgt ccgagtccgt ctgctcctgt ctcctgcacg tactcgtcca
     5221 gcactgtgta gaaaatggtg accacaaaag tgaccacaat gatcagtccg tcggccacct
     5281 cccaaggatt ggtaaagaag ttcttgggtc cgtatgcgaa gatcctcagg ccgagatcca
     5341 gcatgaagta gcagctgagg gccagtgcca tgccgtcgta gaagctctgg gagctctctg
     5401 acttcccagg gagactcaga tcaatgatca tcagaatgat gtccaggaag atgagaaaga
     5461 cgccgaacac ccgcattccc agatggtcga tgacagctct caccctaaac tggactctac
     5521 ccactcctgt ggtaggaatt tccatcctcc cgtcgtccac gttgatgtcg atctggtcca
     5581 gggggatgta ctcgccctcg ctggtgatcc tgctcttctc ctgtcttgtt tcgggcttct
     5641 cgcttggtcc ctgatccacg tcgttccagt ggactttctt cacactctca gattttctag
     5701 gagctgcctt aggtggtgca gtcacgtcgc cgttgattgt cacgtccacc acgttccgca
     5761 tcacagcacc gtccacaccc acgagatcgg caggagggct aaagtctgac ccgtcaaaac
     5821 cttccatggt ggtac
//
