LOCUS       sequence-170999-        6303 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetically encoded voltage sensor ArcLight- codon optimized.
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 ArcLightCo (Q239)-T2A-nls-mCherry
FEATURES             Location/Qualifiers
     source          1..6303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(119..707)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(199..218)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(878..1669,1670..1738))
                     /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     1501..1520
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1739..1843)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4301..4354
                     /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             4376..4396
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4397..5104
                     /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(4539..4557)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4807..4826
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5001..5024
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(5194..5213)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5259..5314
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5313..5332)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5675..5691)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5675..5691)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5688..5710)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5699..5715
                     /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(5723..5729,5730..5747,5748..5753))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5768..5789
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5848..6043
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5894..6029
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6049..6183
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6086..6105)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6140..6159
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6251..6268)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
       61 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
      121 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
      181 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
      241 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
      301 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
      361 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
      421 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
      481 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
      541 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
      601 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
      661 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
      721 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
      781 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
      841 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
      901 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
      961 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     1021 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     1081 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     1141 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     1201 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     1261 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     1321 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     1381 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     1441 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     1501 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     1561 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     1621 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     1681 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     1741 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     1801 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     1861 tgccacctgg gtcgactgca gagggccctc gaccatagcc aattcaatat ggcgtatatg
     1921 gactcgtgcc aattcaatat ggtggatctg gaccccagcc aattcaatat ggcggacttg
     1981 gcaccatgcc aattcaatat ggcggacctg gcactgtgcc aactggggag gggtctactt
     2041 ggcacggtgc caagtttgag gaggggtctt ggccctgtgc caagtccgcc atattgaatt
     2101 ggcatggtgc caataatggc ggccatattg gctatatgcc aggatcaata tataggcaat
     2161 atccaatatg gccctatgcc aatatggcta ttggccaggt tcaatactat gtattggccc
     2221 tatgccatat agtattccat atatgggttt tcctattgac gtagatagcc cctcccaatg
     2281 ggcggtccca tataccatat atggggcttc ctaataccgc ccatagccac tcccccattg
     2341 acgtcaatgg tctctatata tggtctttcc tattgacgtc atatgggcgg tcctattgac
     2401 gtatatggcg cctcccccat tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc
     2461 cattgacgtc aataggacca cccaccattg acgtcaatgg gatggctcat tgcccattca
     2521 tatccgttct cacgccccct attgacgtca atgacggtaa atggcccact tggcagtaca
     2581 tcaatatcta ttaatagtaa cttggcaagt acattactat tggaagtacg ccagggtaca
     2641 ttggcagtac tcccattgac gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca
     2701 tccccattga cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat
     2761 gggcggtagg cgtgcctaat gggaggtcta tataagcaat gctcgtttag ggaacggtac
     2821 caccatggaa ggttttgacg ggtcagactt tagccctcct gccgatctcg tgggtgtgga
     2881 cggtgctgtg atgcggaacg tggtggacgt gacaatcaac ggcgacgtga ctgcaccacc
     2941 taaggcagct cctagaaaat ctgagagtgt gaagaaagtc cactggaacg acgtggatca
     3001 gggaccaagc gagaagcccg aaacaagaca ggagaagagc aggatcacca gcgagggcga
     3061 gtacatcccc ctggaccaga tcgacatcaa cgtggacgac gggaggatgg aaattcctac
     3121 cacaggagtg ggtagagtcc agtttagggt gagagctgtc atcgaccatc tgggaatgcg
     3181 ggtgttcggc gtctttctca tcttcctgga catcattctg atgatcattg atctgagtct
     3241 ccctgggaag tcagagagct cccagagctt ctacgacggc atggcactgg ccctcagctg
     3301 ctacttcatg ctggatctcg gcctgaggat cttcgcatac ggacccaaga acttctttac
     3361 caatccttgg gaggtggccg acggactgat cattgtggtc acttttgtgg tcaccatttt
     3421 ctacacagtg ctggacgagt acgtgcagga gacaggagca gacggactcg gacagctggt
     3481 ggtcctcgct cgactgctcc gagtggtccg actggcaagg atcttttact cccaccagca
     3541 ggggaattca atgtctaaag gggaggaact gttcactggt gtggtcccca ttctggtgga
     3601 gctcgacggc gacgtgaacg gccataagtt ttcagtgagc ggggagggtg aaggcgacgc
     3661 tacttacgga aagctcaccc tgaaattcat ctgcactacc gggaagctgc cagtgccttg
     3721 gcctaccctg gtcacaactc tcacatacgg cgtgcagtgt ttcagtcgct atcctgacca
     3781 tatgaagcga cacgatttct ttaaatctgc tatgccagag ggctacgtgc aggaacggac
     3841 catcttcttt aaggacgacg gcaattataa aactcgcgca gaggtgaagt ttgaaggcga
     3901 caccctggtc aaccgaatcg agctcaaggg aattgacttc aaagaagacg gaaatatcct
     3961 ggggcacaag ctcgagtaca actataacga ccatcaggtg tacatcatgg ccgataagca
     4021 gaaaaacggc attaaggcta acttcaagat ccgccacaac attgaagacg gcggagtgca
     4081 gctggccgat cattaccagc agaacacacc aatcggagac gggcctgtcc tgctcccaga
     4141 taatcactat ctgttcacca catccactct gtctaaggac cccaacgaga aaagagatca
     4201 tatggtgctg ctggagtttg tcactgccga cgggattacc cacggaatgg acgagctgta
     4261 taagttctgc tacgagaacg aggtggcatg cggaagcgga gaaggacggg gtagtctgct
     4321 gacatgcggc gacgtagaag agaatcctgg tcccgccccg gggtcgacca tggtgcccaa
     4381 gaagaagagg aaagtcgtga gcaagggcga ggaggacaac atggccatca tcaaggagtt
     4441 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     4501 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     4561 cggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
     4621 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
     4681 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     4741 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     4801 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     4861 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     4921 ccactacgac gccgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     4981 cgcctacaac gtcaacatca agctggacat cacctcccac aacgaggact acaccatcgt
     5041 ggaacagtac gagcgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     5101 gtaagctagc ggccgcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt
     5161 ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg
     5221 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     5281 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     5341 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     5401 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
     5461 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     5521 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     5581 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc
     5641 cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5701 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5761 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5821 cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc
     5881 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     5941 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     6001 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt
     6061 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     6121 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     6181 atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6241 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6301 cag
//
