LOCUS       sequence-68947-e        6860 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetically encoded voltage indicator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6860)
  AUTHORS   Akemann W, Mutoh H, Perron A, Park YK, Iwamoto Y, Knopfel T
  TITLE     Imaging neural circuit dynamics with a voltage-sensitive fluorescent
            protein.
  JOURNAL   J Neurophysiol. 2012 Oct;108(8):2323-37. doi: 10.1152/jn.00452.2012.
            Epub 2012 Jul 18.
  PUBMED    22815406
REFERENCE   2  (bases 1 to 6860)
  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"; doi: 
            "10.1152/jn.00452.2012"; journalName: "J Neurophysiol"; date: 
            "2012-10"; volume: "108"; issue: "8"; pages: "2323-37"
COMMENT     Sequence Label: pCAG-VSFP Butterfly 1.2
FEATURES             Location/Qualifiers
     source          1..6860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1678..1697
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(3051..3053,3054..3056,3057..3770)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3096..3117)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3357..3376)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3704..3725
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3884..3903)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3949..4004
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4003..4022)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4363..4379)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4363..4379)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4376..4398)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4387..4403
                     /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(4411..4417,4418..4435,4436..4441))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4456..4477
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4536..4731
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4582..4717
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4737..4871
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4774..4793)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4828..4847
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4939..4956)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5110..5698)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5190..5209)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5869..6660,6661..6729))
                     /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     6492..6511
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6730..6834)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcgctagc gccaccatga
     1741 ggagccgctt tgtgaagaaa gatggtcatt gcaatgttca gtttatcaac gtgatggtgt
     1801 ctaagggcga agagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg
     1861 tgaacaacca ccacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc
     1921 agaccatgag aatcaaggcg gtcgagggcg gccctctccc cttcgccttc gacatcctgg
     1981 ctaccagctt catgtacggc agcaaaacct tcatcaacca cacccagggc atccccgact
     2041 tctttaagca gtccttcccc gagggcttca catgggagag agtcaccaca tacgaagacg
     2101 ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg
     2161 tcaagatcag aggggtgaac ttcccatcca acggccctgt gatgcagaag aaaacactcg
     2221 gctgggaggc ctccaccgag accctgtacc ccgctgacgg cggcctggaa ggcagagccg
     2281 acatggccct gaagctcgtg ggcgggggcc acctgatctg caacttgaag accacataca
     2341 gatccaagaa acccgctaag aacctcaaga tgcccggcgt ctactatgtg gacagaagac
     2401 tggaaagaat caaggaggcc gacaaagaga cctacgtcga gcagcacgag gtggctgtgg
     2461 ccagatactg cgacctccct agcaaactgg ggcacagaga ggaacgaata gatatacccg
     2521 agatttcagg tctatggtgg ggcgagaatg aacatggagt ggacgatggg agaatggaga
     2581 tacctactac tggtgtaggt cgcgtccagt ttcgtgtccg agcagtgatt gatcatctag
     2641 ggatgcgagt ctttggagtc ttcctaattt tcttggacat catcctcatg atcattgatc
     2701 tcagtcttcc aggaaaaagt gaatcttcac aatcctttta tgacgggatg gctttggctc
     2761 tttcttgtta tttcatgctg gatttaggat taaggatatt tgcctacggg cccaagaatt
     2821 tcttcaccaa cccctgggag gttgctgatg gtttgattat cgttgtcaca ttcgtcgtca
     2881 cgatatttta cactgtgtta gatgaatacg tgcaagaaac aggagccgat ggtttggggc
     2941 agttggttgt gttggcccgt ttgctgcgtg tggttcgatt agcaagaata ttttattccc
     3001 accaacaaat gaaggcttca agcagaagaa caatatcaca gtgcggccgc atggtgagca
     3061 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     3121 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     3181 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     3241 ccttcggcta cggcctgatg tgcttcgcgc gctaccccga ccacatgaag cagcacgact
     3301 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     3361 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     3421 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     3481 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     3541 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     3601 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
     3661 accagtccaa actgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     3721 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa ggatccgagc
     3781 tcggtaccaa gcttaagtgc ggccgcactc ctcaggtgca ggctgcctat cagaaggtgg
     3841 tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca
     3901 aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat
     3961 ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg
     4021 ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg
     4081 ccatatgctg gctgccatga acaaaggtgg ctataaagag gtcatcagta tatgaaacag
     4141 ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta gatttttttt
     4201 atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta
     4261 ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta
     4321 tgaagatccc tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt
     4381 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4441 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4501 tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata
     4561 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4621 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4681 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact
     4741 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4801 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4861 atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4921 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4981 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5041 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5101 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5161 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5221 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5281 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     5341 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5401 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5461 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5521 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     5581 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5641 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5701 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5761 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5821 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5881 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5941 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6001 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6061 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6121 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6181 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6241 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6301 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6361 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6421 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6481 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6541 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6601 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6661 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6721 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6781 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6841 tccccgaaaa gtgccacctg
//
