LOCUS       sequence-166989-        5833 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 5833)
  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 5833)
  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: hSyn ArcLightCo (Q239)-T2A-nls-mCherry
FEATURES             Location/Qualifiers
     source          1..5833
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..474
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             1963..2016
                     /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             2038..2058
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2059..2766
                     /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(2201..2219)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2469..2488
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2663..2686
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2856..2875)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2921..2976
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2975..2994)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3335..3351)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3335..3351)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3348..3370)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3359..3375
                     /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(3383..3389,3390..3407,3408..3413))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3428..3449
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3508..3703
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3554..3689
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3709..3843
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3746..3765)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3800..3819
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3911..3928)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4082..4670)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4162..4181)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4841..5632,5633..5701))
                     /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     5464..5483
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5702..5806)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgactgca gagggccctg cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg
       61 ggtgggggtg cctacctgac gaccgacccc gacccactgg acaagcaccc aacccccatt
      121 ccccaaattg cgcatcccct atcagagagg gggaggggaa acaggatgcg gcgaggcgcg
      181 tgcgcactgc cagcttcagc accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc
      241 caccgccgcc tcagcactga aggcgcgctg acgtcactcg ccggtccccc gcaaactccc
      301 cttcccggcc accttggtcg cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg
      361 aggcgcgaga taggggggca cgggcgcgac catctgcgct gcggcgccgg cgactcagcg
      421 ctgcctcagt ctgcggtggg cagcggagga gtcgtgtcgt gcctgagagc gcagaccggt
      481 accaccatgg aaggttttga cgggtcagac tttagccctc ctgccgatct cgtgggtgtg
      541 gacggtgctg tgatgcggaa cgtggtggac gtgacaatca acggcgacgt gactgcacca
      601 cctaaggcag ctcctagaaa atctgagagt gtgaagaaag tccactggaa cgacgtggat
      661 cagggaccaa gcgagaagcc cgaaacaaga caggagaaga gcaggatcac cagcgagggc
      721 gagtacatcc ccctggacca gatcgacatc aacgtggacg acgggaggat ggaaattcct
      781 accacaggag tgggtagagt ccagtttagg gtgagagctg tcatcgacca tctgggaatg
      841 cgggtgttcg gcgtctttct catcttcctg gacatcattc tgatgatcat tgatctgagt
      901 ctccctggga agtcagagag ctcccagagc ttctacgacg gcatggcact ggccctcagc
      961 tgctacttca tgctggatct cggcctgagg atcttcgcat acggacccaa gaacttcttt
     1021 accaatcctt gggaggtggc cgacggactg atcattgtgg tcacttttgt ggtcaccatt
     1081 ttctacacag tgctggacga gtacgtgcag gagacaggag cagacggact cggacagctg
     1141 gtggtcctcg ctcgactgct ccgagtggtc cgactggcaa ggatctttta ctcccaccag
     1201 caggggaatt caatgtctaa aggggaggaa ctgttcactg gtgtggtccc cattctggtg
     1261 gagctcgacg gcgacgtgaa cggccataag ttttcagtga gcggggaggg tgaaggcgac
     1321 gctacttacg gaaagctcac cctgaaattc atctgcacta ccgggaagct gccagtgcct
     1381 tggcctaccc tggtcacaac tctcacatac ggcgtgcagt gtttcagtcg ctatcctgac
     1441 catatgaagc gacacgattt ctttaaatct gctatgccag agggctacgt gcaggaacgg
     1501 accatcttct ttaaggacga cggcaattat aaaactcgcg cagaggtgaa gtttgaaggc
     1561 gacaccctgg tcaaccgaat cgagctcaag ggaattgact tcaaagaaga cggaaatatc
     1621 ctggggcaca agctcgagta caactataac gaccatcagg tgtacatcat ggccgataag
     1681 cagaaaaacg gcattaaggc taacttcaag atccgccaca acattgaaga cggcggagtg
     1741 cagctggccg atcattacca gcagaacaca ccaatcggag acgggcctgt cctgctccca
     1801 gataatcact atctgttcac cacatccact ctgtctaagg accccaacga gaaaagagat
     1861 catatggtgc tgctggagtt tgtcactgcc gacgggatta cccacggaat ggacgagctg
     1921 tataagttct gctacgagaa cgaggtggca tgcggaagcg gagaaggacg gggtagtctg
     1981 ctgacatgcg gcgacgtaga agagaatcct ggtcccgccc cggggtcgac catggtgccc
     2041 aagaagaaga ggaaagtcgt gagcaagggc gaggaggaca acatggccat catcaaggag
     2101 ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag
     2161 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     2221 ggcggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag
     2281 gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc
     2341 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac
     2401 tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc
     2461 tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg
     2521 taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc
     2581 ggccactacg acgccgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc
     2641 ggcgcctaca acgtcaacat caagctggac atcacctccc acaacgagga ctacaccatc
     2701 gtggaacagt acgagcgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac
     2761 aagtaagcta gcggccgcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt
     2821 gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat
     2881 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc
     2941 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     3001 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgccatatgc
     3061 tggctgccat gaacaaaggt ggctataaag aggtcatcag tatatgaaac agccccctgc
     3121 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     3181 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     3241 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatgaagatc
     3301 cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     3361 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     3421 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     3481 cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc
     3541 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3601 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3661 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt
     3721 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     3781 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     3841 atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     3901 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3961 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4021 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4081 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4141 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4201 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4261 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4321 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4381 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4441 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4501 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     4561 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4621 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4681 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4741 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4801 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4861 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4921 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4981 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5041 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5101 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5161 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5221 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5281 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5341 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5401 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5461 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5521 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5581 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5641 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5701 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5761 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5821 aagtgccacc tgg
//
