LOCUS       sequence-171002-        7164 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 7164)
  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 7164)
  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 epArcLight
FEATURES             Location/Qualifiers
     source          1..7164
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        268..286
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     268..285
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1053..1769
                     /codon_start=1
                     /product="pH-sensitive mutant of green fluorescent protein 
                     (Miesenbock et al., 1998)"
                     /label=ecliptic pHluorin
                     /note="responds to a drop in pH with a gradual loss 
                     (eclipsing) of excitation at 475 nm, accompanied by a 
                     reduction of excitation at 395 nm"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLLE
                     FVTADGITHGMDELYK"
     primer_bind     complement(1083..1111)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1706..1726
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    1799..1933
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2038..2058)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2043..2372
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2223..2358
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2285..2304
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2421..3446
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             2421..3446
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    3576..3697
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3735..3751)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3735..3751)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3748..3770)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3759..3775
                     /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(3783..3789,3790..3807,3808..3813))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3828..3849
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3966..3983)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4137..4725)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4217..4236)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4896..5687,5688..5756))
                     /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     5519..5538
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5757..5861)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5887..6342)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6024..6045)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6236..6255
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(6413..7164,1..233)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 aatatctatt aatagtaact tggcaagtac attactattg gaagtacgcc agggtacatt
       61 ggcagtactc ccattgacgt caatggcggt aaatggcccg cgatggctgc caagtacatc
      121 cccattgacg tcaatgggga ggggcaatga cgcaaatggg cgttccattg acgtaaatgg
      181 gcggtaggcg tgcctaatgg gaggtctata taagcaatgc tcgtttaggg aaccgccatt
      241 ctgcctgggg acgtcggagc tagcttgatt taggtgacac tatagaatac aagcttatgg
      301 agggattcga cggttcagat tttagtcctc cagctgattt agttggcgtt gacggtgcag
      361 tcatgcggaa cgtcgttgac gtcacgataa atggtgacgt cactgctccg ccgaaagctg
      421 cgccaagaaa aagtgaatcg gtaaagaaag ttcattggaa tgatgtagac caaggaccga
      481 gtgaaaaacc agagacaaga caggaggaac gaatagatat acccgagatt tcaggtctat
      541 ggtggggcga gaatgaacat ggagtggacg atgggagaat ggagatacct actactggtg
      601 taggtcgcgt ccagtttcgt gtccgagcag tgattgatca tctagggatg cgagtctttg
      661 gagtcttcct aattttcttg gacatcatcc tcatgatcat tgatctcagt cttccaggaa
      721 aaagtgaatc ttcacaatcc ttttatgacg ggatggcttt ggctctttct tgttatttca
      781 tgctggattt aggattaagg atatttgcct acgggcccaa gaatttcttc accaacccct
      841 gggaggttgc tgatggtttg attatcgttg tcacattcgt cgtcacgata ttttacactg
      901 tgttagatga atacgtgcaa gaaacaggag ccgatggttt ggggcaattg gttgtgttgg
      961 cccgtttgct gcgtgtggtt cgattagcaa gaatatttta ttcccaccaa caaatgaagg
     1021 cttcaagcag aagaacaata tcaggggatc ccatgagtaa aggagaagaa cttttcactg
     1081 gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca
     1141 gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta
     1201 ctggaaaact acctgttcca tggccaacac ttgtcactac tttctcttat ggtgttcaat
     1261 gcttttcaag atacccagat catatgaaac ggcatgactt tttcaagagt gccatgcccg
     1321 aaggttatgt acaggaaaga actatatttt tcaaagatga cgggaactac aagacacgtg
     1381 ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt
     1441 ttaaagaaga tggaaacatt cttggacaca aattggaata caactataac gatcaccagg
     1501 tgtacatcat ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa attagacaca
     1561 acattgaaga tggaggcgtt caactagcag accattatca acaaaatact ccaattggcg
     1621 atgggcccgt ccttttacca gacaaccatt acctgtttac aacttctact ctttcgaaag
     1681 atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct gatgggatta
     1741 cacatggcat ggatgaacta tacaaatgat ctagaactat agtgagtcgt attacgtaga
     1801 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     1861 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     1921 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     1981 gtgggaggtt ttttaattga tatccgcggc gccaatgcat tgggcccggt accctgtgga
     2041 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2101 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2161 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2221 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2281 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     2341 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     2401 tcggatctga tcagcacgtg atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt
     2461 ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat
     2521 ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg
     2581 ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga
     2641 ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc atctcccgcc
     2701 gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc
     2761 cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg
     2821 gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga
     2881 ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg
     2941 tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc
     3001 tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg
     3061 tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct
     3121 tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc
     3181 cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac
     3241 tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg
     3301 acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg
     3361 cggccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca
     3421 gcactcgtcc gagggcaaag gaatagcacg tgctacgaga tttcgattcc accgccgcct
     3481 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     3541 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     3601 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3661 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtacgcgc
     3721 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3781 cacaacatac gagccgggag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3841 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3901 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3961 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4021 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4081 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4141 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4201 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4261 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4321 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4381 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4441 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4501 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4561 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4621 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4681 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4741 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4801 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4861 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4921 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4981 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5041 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5101 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5161 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5221 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5281 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5341 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5401 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5461 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5521 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5581 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5641 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5701 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5761 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5821 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5881 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     5941 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     6001 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     6061 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     6121 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     6181 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     6241 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     6301 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     6361 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagtcgaccg
     6421 ccaattcaat atggcgtata tggactcatg ccaattcaat atggtggatc tggacctgtg
     6481 ccaattcaat atggcgtata tggactcgtg ccaattcaat atggtggatc tggaccccag
     6541 ccaattcaat atggcggact tggcaccatg ccaattcaat atggcggacc tggcactgtg
     6601 ccaactgggg aggggtctac ttggcacggt gccaagtttg aggaggggtc ttggccctgt
     6661 gccaagtccg ccatattgaa ttggcatggt gccaataatg gcggccatat tggctatatg
     6721 ccaggatcaa tatataggca atatccaata tggccctatg ccaatatggc tattggccag
     6781 gttcaatact atgtattggc cctatgccat atagtattcc atatatgggt tttcctattg
     6841 acgtagatag cccctcccaa tgggcggtcc catataccat atatggggct tcctaatacc
     6901 gcccatagcc actcccccat tgacgtcaat ggtctctata tatggtcttt cctattgacg
     6961 tcatatgggc ggtcctattg acgtatatgg cgcctccccc attgacgtca attacggtaa
     7021 atggcccgcc tggctcaatg cccattgacg tcaataggac cacccaccat tgacgtcaat
     7081 gggatggctc attgcccatt catatccgtt ctcacgcccc ctattgacgt caatgacggt
     7141 aaatggccca cttggcagta catc
//
