LOCUS       sequence-83736-e        6949 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 6949)
  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 6949)
  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: Chicken ArcLight A173
FEATURES             Location/Qualifiers
     source          1..6949
                     /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             602..1318
                     /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(632..660)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1255..1275
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    1348..1482
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1587..1607)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1592..1921
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1772..1907
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1834..1853
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1970..2995
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             1970..2995
                     /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    3125..3246
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3284..3300)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3284..3300)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3297..3319)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3308..3324
                     /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(3332..3338,3339..3356,3357..3362))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3377..3398
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3515..3532)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3686..4274)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3766..3785)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4445..5236,5237..5305))
                     /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     5068..5087
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5306..5410)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5436..5891)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5573..5594)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5785..5804
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5965..6949
                     /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 atggagacga ctgtgaggta tgaacagggg tcagagctca ctaaaacttc
      121 gagctctcca acagcagatg agcccacgat aaagattgat gatggtcgtg atgagggtaa
      181 tgaacaagac agctgttcca ataccattag gagaaaaatt tccccgtttg tgatgtcatt
      241 tggattcaga gtatttggag ttgtgcttat cattgtagac atcatagtgg tgattgtgga
      301 tctggccatc agtgagaaga aaagaggcat tagagagatt cttgaaggtg tttccctggc
      361 tatagcactc ttcttccttg ttgatgttct catgagagtg tttgttgaag gcttcaagaa
      421 ctatttccgg tccaaactga atactttgga tgcagtcata gtagtgggca ctctgctaat
      481 taatatgacc tactccttct ctgaccttgc tgccacagat cagatgccgc agatggttac
      541 tcttcttcga gttctgagaa ttgttatctt aataagaata tttcgcctgg ctggggatcc
      601 catgagtaaa ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg
      661 tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg caacatacgg
      721 aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact
      781 tgtcactact ttaacttatg gtgttcaatg cttttcaaga tacccagatc atatgaaacg
      841 gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaaagaa ctatattttt
      901 caaagatgac gggaactaca agacacgtgc tgaagtcaag tttgaaggtg atacccttgt
      961 taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc ttggacacaa
     1021 attggaatac aactataacg atcaccaggt gtacatcatg gcagacaaac aaaagaatgg
     1081 aatcaaagct aacttcaaaa ttagacacaa cattgaagat ggaggcgttc aactagcaga
     1141 ccattatcaa caaaatactc caattggcga tgggcccgtc cttttaccag acaaccatta
     1201 cctgtttaca acttctactc tttcgaaaga tcccaacgaa aagagagacc acatggtcct
     1261 tcttgagttt gtaacagctg atgggattac acatggcatg gatgaactat acaaatgatc
     1321 tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat tgatgagttt
     1381 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     1441 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     1501 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaattgat atccgcggcg
     1561 ccaatgcatt gggcccggta ccctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     1621 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     1681 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     1741 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     1801 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     1861 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     1921 aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtga tgaaaaagcc
     1981 tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga
     2041 cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg
     2101 tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta
     2161 tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaattcag
     2221 cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc
     2281 tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc
     2341 ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata
     2401 cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac
     2461 tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg
     2521 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt
     2581 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga
     2641 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca
     2701 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc
     2761 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga
     2821 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt
     2881 cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt
     2941 actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aatagcacgt
     3001 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3061 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     3121 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3181 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3241 atcttatcat gtctgtatac cgtacgcgct tggcgtaatc atggtcatag ctgtttcctg
     3301 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3361 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3421 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3481 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3541 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3601 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3661 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3721 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3781 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3841 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3901 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3961 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4021 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4081 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     4141 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4201 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4261 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4321 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4381 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4441 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4501 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4561 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4621 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4681 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4741 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4801 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4861 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4921 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4981 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5041 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5101 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5161 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5221 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5281 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5341 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5401 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
     5461 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     5521 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     5581 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     5641 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     5701 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     5761 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     5821 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     5881 acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     5941 ggcctcttcg ctattacgcc agtcgaccat agccaattca atatggcgta tatggactca
     6001 tgccaattca atatggtgga tctggacctg tgccaattca atatggcgta tatggactcg
     6061 tgccaattca atatggtgga tctggacccc agccaattca atatggcgga cttggcacca
     6121 tgccaattca atatggcgga cttggcactg tgccaactgg ggaggggtct acttggcacg
     6181 gtgccaagtt tgaggagggg tcttggccct gtgccaagtc cgccatattg aattggcatg
     6241 gtgccaataa tggcggccat attggctata tgccaggatc aatatatagg caatatccaa
     6301 tatggcccta tgccaatatg gctattggcc aggttcaata ctatgtattg gccctatgcc
     6361 atatagtatt ccatatatgg gttttcctat tgacgtagat agcccctccc aatgggcggt
     6421 cccatatacc atatatgggg cttcctaata ccgcccatag ccactccccc attgacgtca
     6481 atggtctcta tatatggtct ttcctattga cgtcatatgg gcggtcctat tgacgtatat
     6541 ggcgcctccc ccattgacgt caattacggt aaatggcccg cctggctcaa tgcccattga
     6601 cgtcaatagg accacccacc attgacgtca atgggatggc tcattgccca ttcatatccg
     6661 ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc cacttggcag tacatcaata
     6721 tctattaata gtaacttggc aagtacatta ctattggaag gacgccaggg tacattggca
     6781 gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag tacatcccca
     6841 ttgacgtcaa tggggagggg caatgacgca aatgggcgtt ccattgacgt aaatgggcgg
     6901 taggcgtgcc taatgggagg tctatataag caatgctcgt ttagggaac
//
