LOCUS       sequence-83742-e        6955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetically encoded voltage sensor ArcLight with improved kinetics .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6955)
  AUTHORS   Han Z, Jin L, Platisa J, Cohen LB, Baker BJ, Pieribone VA
  TITLE     Fluorescent protein voltage probes derived from ArcLight that 
            respond to membrane voltage changes with fast kinetics.
  JOURNAL   PLoS One. 2013 Nov 27;8(11):e81295. doi: 
            10.1371/journal.pone.0081295. eCollection 2013.
  PUBMED    24312287
REFERENCE   2  (bases 1 to 6955)
  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: "PLoS One.";
            date: "2013-11-27"; volume: "8(11):e81295. doi"; pages: " 
            10.1371/journal.pone.0081295. eCollection 2013"
COMMENT     Sequence Label: Zebrafish ArcLight Q175
FEATURES             Location/Qualifiers
     source          1..6955
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      65..74
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             608..1324
                     /codon_start=1
                     /product="pH-sensitive mutant of green fluorescent protein 
                     (Ng et al., 2002)"
                     /label=superecliptic pHluorin
                     /note="excited strongly by blue light at neutral pH but 
                     minimally fluorescent at acidic pH"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLLE
                     FVTADGITHGMDELYK"
     primer_bind     complement(638..666)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1261..1281
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    1354..1488
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1593..1613)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1598..1927
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1778..1913
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1840..1859
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1976..3001
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             1976..3001
                     /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    3131..3252
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3290..3306)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3290..3306)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3303..3325)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3314..3330
                     /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(3338..3344,3345..3362,3363..3368))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3383..3404
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3521..3538)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3692..4280)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3772..3791)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4451..5242,5243..5311))
                     /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     5074..5093
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5312..5416)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5442..5897)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5579..5600)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5791..5810
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5971..6955
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagctag cttgatttag gtgacactat agaatacaag
       61 cttcgccacc atggagacgt ctgtgcattt taaccctggg ttagattcca aagaagttaa
      121 tggtaattcg gtgaaagaag aggcagaggt gcagatcgat gatggaaagg aagaaaccaa
      181 agacccagac acaatgtacc atcaggtccg gaaaaagata actccttttg tgatgtcctt
      241 tggttttcgt gtttttggcc tggtactgat catcttggac atcattatgg tcattgtgga
      301 cttgtctctt tctgagaaga gtcgtgatgt tggaggtgca ccggagactg tttctctggt
      361 catctctttc ttttttctca tcgatgtatt gctccgtgtt tatgtggagg gattcaaggt
      421 gtatttcagc tcaaaactga atatagtaga tgcttgtatt gtggtcatta ctctggtggt
      481 caccatgatc tacgcattct ccgatttctc aggagccagt ctgattcccc aggtggtgac
      541 attcctgagg tctctgagga tcctaattct ggtacgcatc ttcagactgg cctcgcaggg
      601 agatcccatg agtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt
      661 agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac
      721 atacggaaaa cttaccctta aatttatttg cactactgga aaactacctg ttccatggcc
      781 aacacttgtc actactttaa cttatggtgt tcaatgcttt tcaagatacc cagatcatat
      841 gaaacggcat gactttttca agagtgccat gcccgaaggt tatgtacagg aaagaactat
      901 atttttcaaa gatgacggga actacaagac acgtgctgaa gtcaagtttg aaggtgatac
      961 ccttgttaat agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg
     1021 acacaaattg gaatacaact ataacgatca ccaggtgtac atcatggcag acaaacaaaa
     1081 gaatggaatc aaagctaact tcaaaattag acacaacatt gaagatggag gcgttcaact
     1141 agcagaccat tatcaacaaa atactccaat tggcgatggg cccgtccttt taccagacaa
     1201 ccattacctg tttacaactt ctactctttc gaaagatccc aacgaaaaga gagaccacat
     1261 ggtccttctt gagtttgtaa cagctgatgg gattacacat ggcatggatg aactatacaa
     1321 atgatctaga actatagtga gtcgtattac gtagatccag acatgataag atacattgat
     1381 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     1441 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     1501 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta attgatatcc
     1561 gcggcgccaa tgcattgggc ccggtaccct gtggaatgtg tgtcagttag ggtgtggaaa
     1621 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     1681 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     1741 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     1801 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     1861 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     1921 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgatgaa
     1981 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
     2041 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
     2101 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
     2161 tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga
     2221 attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga
     2281 cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat
     2341 cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg
     2401 tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg
     2461 gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat
     2521 gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa
     2581 caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt
     2641 cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat
     2701 ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct
     2761 ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa
     2821 tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg
     2881 gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt
     2941 agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata
     3001 gcacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     3061 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     3121 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3181 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3241 caatgtatct tatcatgtct gtataccgta cgcgcttggc gtaatcatgg tcatagctgt
     3301 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     3361 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     3421 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     3481 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     3541 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3601 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3661 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3721 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3781 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3841 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3901 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3961 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4021 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4081 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     4141 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4201 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4261 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4321 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4381 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4441 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4501 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4561 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4621 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4681 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4741 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4801 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4861 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4921 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4981 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5041 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5101 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5161 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5221 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5281 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5341 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5401 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     5461 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     5521 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     5581 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     5641 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     5701 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     5761 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     5821 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     5881 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
     5941 ggtgcgggcc tcttcgctat tacgccagtc gaccatagcc aattcaatat ggcgtatatg
     6001 gactcatgcc aattcaatat ggtggatctg gacctgtgcc aattcaatat ggcgtatatg
     6061 gactcgtgcc aattcaatat ggtggatctg gaccccagcc aattcaatat ggcggacttg
     6121 gcaccatgcc aattcaatat ggcggacttg gcactgtgcc aactggggag gggtctactt
     6181 ggcacggtgc caagtttgag gaggggtctt ggccctgtgc caagtccgcc atattgaatt
     6241 ggcatggtgc caataatggc ggccatattg gctatatgcc aggatcaata tataggcaat
     6301 atccaatatg gccctatgcc aatatggcta ttggccaggt tcaatactat gtattggccc
     6361 tatgccatat agtattccat atatgggttt tcctattgac gtagatagcc cctcccaatg
     6421 ggcggtccca tataccatat atggggcttc ctaataccgc ccatagccac tcccccattg
     6481 acgtcaatgg tctctatata tggtctttcc tattgacgtc atatgggcgg tcctattgac
     6541 gtatatggcg cctcccccat tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc
     6601 cattgacgtc aataggacca cccaccattg acgtcaatgg gatggctcat tgcccattca
     6661 tatccgttct cacgccccct attgacgtca atgacggtaa atggcccact tggcagtaca
     6721 tcaatatcta ttaatagtaa cttggcaagt acattactat tggaaggacg ccagggtaca
     6781 ttggcagtac tcccattgac gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca
     6841 tccccattga cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat
     6901 gggcggtagg cgtgcctaat gggaggtcta tataagcaat gctcgtttag ggaac
//
