LOCUS       sequence-166999-        6361 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 6361)
  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 6361)
  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: CMV Marina-T2A-nls-mCherry
FEATURES             Location/Qualifiers
     source          1..6361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..1005
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     CDS             2491..2544
                     /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             2566..2586
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2587..3294
                     /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(2729..2747)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2997..3016
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3191..3214
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(3384..3403)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3449..3504
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3503..3522)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3863..3879)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3863..3879)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3876..3898)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3887..3903
                     /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(join(3911..3917,3918..3935,3936..3941))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3956..3977
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4036..4231
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4082..4217
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4237..4371
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4274..4293)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4328..4347
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4439..4456)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4610..5198)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4690..4709)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5369..6160,6161..6229))
                     /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     5992..6011
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6230..6334)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgactgca gagggccctc gaccatagcc aattcaatat ggcgtatatg gactcatgcc
       61 aattcaatat ggtggatctg gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc
      121 aattcaatat ggtggatctg gaccccagcc aattcaatat ggcggacttg gcaccatgcc
      181 aattcaatat ggcggacttg gcactgtgcc aactggggag gggtctactt ggcacggtgc
      241 caagtttgag gaggggtctt ggccctgtgc caagtccgcc atattgaatt ggcatggtgc
      301 caataatggc ggccatattg gctatatgcc aggatcaata tataggcaat atccaatatg
      361 gccctatgcc aatatggcta ttggccaggt tcaatactat gtattggccc tatgccatat
      421 agtattccat atatgggttt tcctattgac gtagatagcc cctcccaatg ggcggtccca
      481 tataccatat atggggcttc ctaataccgc ccatagccac tcccccattg acgtcaatgg
      541 tctctatata tggtctttcc tattgacgtc atatgggcgg tcctattgac gtatatggcg
      601 cctcccccat tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc cattgacgtc
      661 aataggacca cccaccattg acgtcaatgg gatggctcat tgcccattca tatccgttct
      721 cacgccccct attgacgtca atgacggtaa atggcccact tggcagtaca tcaatatcta
      781 ttaatagtaa cttggcaagt acattactat tggaaggacg ccagggtaca ttggcagtac
      841 tcccattgac gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca tccccattga
      901 cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat gggcggtagg
      961 cgtgcctaat gggaggtcta tataagcaat gctcgtttag ggaacggtac caccatggaa
     1021 ggttttgacg ggtcagactt tagccctcct gccgatctcg tgggtgtgga cggtgctgtg
     1081 atgcggaacg tggtggacgt gacaatcaac ggcgacgtga ctgcaccacc taaggcagct
     1141 cctagaaaat ctgagagtgt gaagaaagtc cactggaacg acgtggatca gggaccaagc
     1201 gagaagcccg aaacaagaca ggagaagagc aggatcacca gcgagggcga gtacatcccc
     1261 ctggaccaga tcgacatcaa cgtggacgac gggaggatgg aaattcctac cacaggagtg
     1321 ggtagagtcc agtttagggt gagagctgtc atcgaccatc tgggaatgcg ggtgttcggc
     1381 gtctttctca tcttcctgga catcattctg atgatcattg atctgagtct ccctgggaag
     1441 tcagagagct cccagagctt ctacgacggc atggcactgg ccctcagctg ctacttcatg
     1501 ctggatctcg gcctgaggat cttcgcatac ggacccaaga acttctttac caatccttgg
     1561 gaggtggccg acggactgat cattgtggtc acttttgtgg tcaccatttt ctacacagtg
     1621 ctggacgagt acgtgcagga gacaggagca gacggactcg gacagctggt ggtcctcgct
     1681 cgactgctcc gagtggtccg actggcaagg atcttttact cccaccagca ggggaattca
     1741 atgtctaaag gggaggaact gttcactggt gtggtcccca ttctggtgga gctcgacggc
     1801 gacgtgaacg gccataagtt ttcagtgagc ggggagggtg aaggcgacgc tacttacgga
     1861 aagctcaccc tgaaattcat ctgcactacc gggaagctgc cagtgccttg gcctaccctg
     1921 gtcacaactc tcacatacgg cgtgcagtgt ttcagtcgct atcctgacca tatgaagcga
     1981 cacgatttct ttaaatctgc tatgccagag ggctacgtgc aggaacggac catcttcttt
     2041 aaggacgacg gcaattataa aactcgcgca gaggtgaagt ttgaaggcga caccctggtc
     2101 aaccgaatcg agctcaaggg aattgacttc aaagaagacg gaaatatcct ggggcacaag
     2161 ctcgagtaca actataacgc cgctcaggtg tacatcatgg ccgataagca gaaaaacggc
     2221 attaaggcta acttcaagat ccgccacaac attgaagacg gcggagtgca gctggccgat
     2281 cattaccagc agaacacacc aatcggagac gggcctgtcc tgctcccaga taatcacgtt
     2341 ctgttcacca catccactct gtctaaggac cccaacgaga aaagagatca tatggtgctg
     2401 ctggagtttg tcactgccga cgggattacc cacggaatgg acgagctgta taagttctgc
     2461 tacgagaacg aggtggcatg cggaagcgga gaaggacggg gtagtctgct gacatgcggc
     2521 gacgtagaag agaatcctgg tcccgccccg gggtcgacca tggtgcccaa gaagaagagg
     2581 aaagtcgtga gcaagggcga ggaggacaac atggccatca tcaaggagtt catgcgcttc
     2641 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     2701 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg
     2761 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     2821 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     2881 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     2941 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     3001 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     3061 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     3121 gccgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     3181 gtcaacatca agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     3241 gagcgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaagctagc
     3301 ggccgcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc
     3361 ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg
     3421 aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg
     3481 tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa
     3541 catcagaatg agtatttggt ttagagtttg gcaacatatg ccatatgctg gctgccatga
     3601 acaaaggtgg ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt
     3661 attccataga aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt
     3721 tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct
     3781 ctcctgacta ctcccagtca tagctgtccc tcttctctta tgaagatccc tcgacctgca
     3841 gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3901 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3961 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4021 cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4081 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4141 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4201 aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat
     4261 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4321 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt
     4381 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4441 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4501 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4561 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4621 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4681 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4741 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4801 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4861 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4921 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4981 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5041 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5101 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5161 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5221 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5281 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5341 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5401 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5461 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5521 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5581 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5641 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5701 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5761 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5821 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5881 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5941 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6001 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6061 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6121 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6181 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6241 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6301 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6361 g
//
