LOCUS       sequence-83744-e        6952 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 6952)
  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 6952)
  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 S174
FEATURES             Location/Qualifiers
     source          1..6952
                     /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             605..1321
                     /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(635..663)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1258..1278
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    1351..1485
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1590..1610)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1595..1924
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1775..1910
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1837..1856
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1973..2998
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             1973..2998
                     /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    3128..3249
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3287..3303)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3287..3303)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3300..3322)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3311..3327
                     /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(3335..3341,3342..3359,3360..3365))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3380..3401
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3518..3535)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3689..4277)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3769..3788)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4448..5239,5240..5308))
                     /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     5071..5090
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5309..5413)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5439..5894)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5576..5597)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5788..5807
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5968..6952
                     /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 cttcagggga
      601 tcccatgagt aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga
      661 tggtgatgtt aatgggcaca aattttctgt cagtggagag ggtgaaggtg atgcaacata
      721 cggaaaactt acccttaaat ttatttgcac tactggaaaa ctacctgttc catggccaac
      781 acttgtcact actttaactt atggtgttca atgcttttca agatacccag atcatatgaa
      841 acggcatgac tttttcaaga gtgccatgcc cgaaggttat gtacaggaaa gaactatatt
      901 tttcaaagat gacgggaact acaagacacg tgctgaagtc aagtttgaag gtgataccct
      961 tgttaataga atcgagttaa aaggtattga ttttaaagaa gatggaaaca ttcttggaca
     1021 caaattggaa tacaactata acgatcacca ggtgtacatc atggcagaca aacaaaagaa
     1081 tggaatcaaa gctaacttca aaattagaca caacattgaa gatggaggcg ttcaactagc
     1141 agaccattat caacaaaata ctccaattgg cgatgggccc gtccttttac cagacaacca
     1201 ttacctgttt acaacttcta ctctttcgaa agatcccaac gaaaagagag accacatggt
     1261 ccttcttgag tttgtaacag ctgatgggat tacacatggc atggatgaac tatacaaatg
     1321 atctagaact atagtgagtc gtattacgta gatccagaca tgataagata cattgatgag
     1381 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     1441 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     1501 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaatt gatatccgcg
     1561 gcgccaatgc attgggcccg gtaccctgtg gaatgtgtgt cagttagggt gtggaaagtc
     1621 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     1681 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     1741 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     1801 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     1861 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     1921 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgatgaaaaa
     1981 gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc
     2041 cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg
     2101 gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt
     2161 ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt
     2221 cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct
     2281 gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc
     2341 tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca
     2401 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca
     2461 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
     2521 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
     2581 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
     2641 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
     2701 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg
     2761 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
     2821 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
     2881 tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga
     2941 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagca
     3001 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     3061 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     3121 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3181 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3241 tgtatcttat catgtctgta taccgtacgc gcttggcgta atcatggtca tagctgtttc
     3301 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     3361 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     3421 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3481 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3541 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3601 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3661 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3721 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3781 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3841 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3901 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3961 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4021 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4081 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     4141 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4201 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4261 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4321 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4381 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4441 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4501 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4561 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4621 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4681 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4741 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4801 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4861 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4921 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4981 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5041 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5101 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5161 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5221 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5281 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5341 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5401 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     5461 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     5521 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     5581 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     5641 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     5701 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     5761 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     5821 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     5881 gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     5941 gcgggcctct tcgctattac gccagtcgac catagccaat tcaatatggc gtatatggac
     6001 tcatgccaat tcaatatggt ggatctggac ctgtgccaat tcaatatggc gtatatggac
     6061 tcgtgccaat tcaatatggt ggatctggac cccagccaat tcaatatggc ggacttggca
     6121 ccatgccaat tcaatatggc ggacttggca ctgtgccaac tggggagggg tctacttggc
     6181 acggtgccaa gtttgaggag gggtcttggc cctgtgccaa gtccgccata ttgaattggc
     6241 atggtgccaa taatggcggc catattggct atatgccagg atcaatatat aggcaatatc
     6301 caatatggcc ctatgccaat atggctattg gccaggttca atactatgta ttggccctat
     6361 gccatatagt attccatata tgggttttcc tattgacgta gatagcccct cccaatgggc
     6421 ggtcccatat accatatatg gggcttccta ataccgccca tagccactcc cccattgacg
     6481 tcaatggtct ctatatatgg tctttcctat tgacgtcata tgggcggtcc tattgacgta
     6541 tatggcgcct cccccattga cgtcaattac ggtaaatggc ccgcctggct caatgcccat
     6601 tgacgtcaat aggaccaccc accattgacg tcaatgggat ggctcattgc ccattcatat
     6661 ccgttctcac gccccctatt gacgtcaatg acggtaaatg gcccacttgg cagtacatca
     6721 atatctatta atagtaactt ggcaagtaca ttactattgg aaggacgcca gggtacattg
     6781 gcagtactcc cattgacgtc aatggcggta aatggcccgc gatggctgcc aagtacatcc
     6841 ccattgacgt caatggggag gggcaatgac gcaaatgggc gttccattga cgtaaatggg
     6901 cggtaggcgt gcctaatggg aggtctatat aagcaatgct cgtttaggga ac
//
