LOCUS       sequence-170988-        6303 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 6303)
  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 6303)
  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..6303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    149..283
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(186..205)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     240..259
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(351..368)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(522..1110)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(602..621)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1281..2072,2073..2141))
                     /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     1904..1923
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2142..2246)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4704..4757
                     /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             4779..4799
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4800..5507
                     /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(4942..4960)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5210..5229
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5404..5427
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(5597..5616)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5662..5717
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5716..5735)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6078..6094)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6078..6094)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6091..6113)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6102..6118
                     /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(6126..6132,6133..6150,6151..6156))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6171..6192
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6251..6303,1..143)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      join(6297..6303,1..129)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
       61 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
      121 ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata atggttacaa
      181 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
      241 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgctgca ttaatgaatc
      301 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
      361 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
      421 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
      481 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
      541 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      601 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      661 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      721 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      781 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      841 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      901 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      961 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1021 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     1081 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     1141 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     1201 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     1261 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     1321 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     1381 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     1441 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1501 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1561 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1621 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1681 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1741 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1801 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1861 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1921 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1981 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     2041 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     2101 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     2161 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     2221 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgggtcgact
     2281 gcagagggcc ctcgaccata gccaattcaa tatggcgtat atggactcgt gccaattcaa
     2341 tatggtggat ctggacccca gccaattcaa tatggcggac ttggcaccat gccaattcaa
     2401 tatggcggac ctggcactgt gccaactggg gaggggtcta cttggcacgg tgccaagttt
     2461 gaggaggggt cttggccctg tgccaagtcc gccatattga attggcatgg tgccaataat
     2521 ggcggccata ttggctatat gccaggatca atatataggc aatatccaat atggccctat
     2581 gccaatatgg ctattggcca ggttcaatac tatgtattgg ccctatgcca tatagtattc
     2641 catatatggg ttttcctatt gacgtagata gcccctccca atgggcggtc ccatatacca
     2701 tatatggggc ttcctaatac cgcccatagc cactccccca ttgacgtcaa tggtctctat
     2761 atatggtctt tcctattgac gtcatatggg cggtcctatt gacgtatatg gcgcctcccc
     2821 cattgacgtc aattacggta aatggcccgc ctggctcaat gcccattgac gtcaatagga
     2881 ccacccacca ttgacgtcaa tgggatggct cattgcccat tcatatccgt tctcacgccc
     2941 cctattgacg tcaatgacgg taaatggccc acttggcagt acatcaatat ctattaatag
     3001 taacttggca agtacattac tattggaagt acgccagggt acattggcag tactcccatt
     3061 gacgtcaatg gcggtaaatg gcccgcgatg gctgccaagt acatccccat tgacgtcaat
     3121 ggggaggggc aatgacgcaa atgggcgttc cattgacgta aatgggcggt aggcgtgcct
     3181 aatgggaggt ctatataagc aatgctcgtt tagggaacgg taccaccatg gaaggttttg
     3241 acgggtcaga ctttagccct cctgccgatc tcgtgggtgt ggacggtgct gtgatgcgga
     3301 acgtggtgga cgtgacaatc aacggcgacg tgactgcacc acctaaggca gctcctagaa
     3361 aatctgagag tgtgaagaaa gtccactgga acgacgtgga tcagggacca agcgagaagc
     3421 ccgaaacaag acaggagaag agcaggatca ccagcgaggg cgagtacatc cccctggacc
     3481 agatcgacat caacgtggac gacgggagga tggaaattcc taccacagga gtgggtagag
     3541 tccagtttag ggtgagagct gtcatcgacc atctgggaat gcgggtgttc ggcgtctttc
     3601 tcatcttcct ggacatcatt ctgatgatca ttgatctgag tctccctggg aagtcagaga
     3661 gctcccagag cttctacgac ggcatggcac tggccctcag ctgctacttc atgctggatc
     3721 tcggcctgag gatcttcgca tacggaccca agaacttctt taccaatcct tgggaggtgg
     3781 ccgacggact gatcattgtg gtcacttttg tggtcaccat tttctacaca gtgctggacg
     3841 agtacgtgca ggagacagga gcagacggac tcggacagct ggtggtcctc gctcgactgc
     3901 tccgagtggt ccgactggca aggatctttt actcccacca gcaggggaat tcaatgtcta
     3961 aaggggagga actgttcact ggtgtggtcc ccattctggt ggagctcgac ggcgacgtga
     4021 acggccataa gttttcagtg agcggggagg gtgaaggcga cgctacttac ggaaagctca
     4081 ccctgaaatt catctgcact accgggaagc tgccagtgcc ttggcctacc ctggtcacaa
     4141 ctctcacata cggcgtgcag tgtttcagtc gctatcctga ccatatgaag cgacacgatt
     4201 tctttaaatc tgctatgcca gagggctacg tgcaggaacg gaccatcttc tttaaggacg
     4261 acggcaatta taaaactcgc gcagaggtga agtttgaagg cgacaccctg gtcaaccgaa
     4321 tcgagctcaa gggaattgac ttcaaagaag acggaaatat cctggggcac aagctcgagt
     4381 acaactataa cgccgctcag gtgtacatca tggccgataa gcagaaaaac ggcattaagg
     4441 ctaacttcaa gatccgccac aacattgaag acggcggagt gcagctggcc gatcattacc
     4501 agcagaacac accaatcgga gacgggcctg tcctgctccc agataatcac gttctgttca
     4561 ccacatccac tctgtctaag gaccccaacg agaaaagaga tcatatggtg ctgctggagt
     4621 ttgtcactgc cgacgggatt acccacggaa tggacgagct gtataagttc tgctacgaga
     4681 acgaggtggc atgcggaagc ggagaaggac ggggtagtct gctgacatgc ggcgacgtag
     4741 aagagaatcc tggtcccgcc ccggggtcga ccatggtgcc caagaagaag aggaaagtcg
     4801 tgagcaaggg cgaggaggac aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
     4861 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     4921 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg
     4981 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     5041 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     5101 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
     5161 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     5221 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     5281 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgccgagg
     5341 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     5401 tcaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgagcgcg
     5461 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaagct agcggccgca
     5521 ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc
     5581 acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     5641 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     5701 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
     5761 atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag
     5821 gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc
     5881 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt
     5941 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct
     6001 gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac ctgcagccca
     6061 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     6121 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     6181 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6241 cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     6301 cgc
//
