LOCUS       sequence-270808-        7454 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a genetically-encoded voltage indicator nirButterfly .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7454)
  AUTHORS   Monakhov MV, Matlashov ME, Colavita M, Song C, Shcherbakova DM, 
            Antic SD, Verkhusha VV, Knopfel T
  TITLE     Screening and Cellular Characterization of Genetically Encoded 
            Voltage Indicators Based on Near-Infrared Fluorescent Proteins.
  JOURNAL   ACS Chem Neurosci. 2020 Nov 4;11(21):3523-3531. doi: 
            10.1021/acschemneuro.0c00046. Epub 2020 Oct 16.
  PUBMED    33063984
REFERENCE   2  (bases 1 to 7454)
  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: 
            "10.1021/acschemneuro.0c00046"; journalName: "ACS Chem Neurosci"; 
            date: "2020-11-4- 4"; volume: "11"; issue: "21"; pages: "3523-3531"
COMMENT     Sequence Label: pCAG-nirButterfly
FEATURES             Location/Qualifiers
     source          1..7454
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        36..311
                     /label=chicken beta-actin promoter
     intron          312..1328
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1252..1274
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1336..1355
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1383..1392
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1704..2651
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 720 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP720
                     /note="derived from Rhodopseudomonas palustris RpBphP2"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGSVIAEDRCAEVESKLGLHYPASFIPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPNHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACKRVAERLATQIGVMEE"
     primer_bind     complement(4126..4145)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4191..4246
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4245..4264)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4607..4623)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4607..4623)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4620..4642)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4631..4647
                     /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(4655..4661,4662..4679,4680..4685))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4700..4721
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4780..4975
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4826..4961
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4981..5115
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5018..5037)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5072..5091
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5183..5200)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5354..5942)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5434..5453)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6113..6904,6905..6973))
                     /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     6736..6755
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6974..7078)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(7109..7454,1..34)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt
       61 cactctcccc atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt
      121 attttgtgca gcgatggggg cggggggggg ggggncgcgc gccaggcggg gcggggcggg
      181 gcgaggggcg gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc
      241 tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg
      301 cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct
      361 cgcgccgccc gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg
      421 gcccttctcc tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt
      481 ggctgcgtga aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg
      541 ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg
      601 gctgtgagcg ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga
      661 gcgcggccgg gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg
      721 tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac
      781 cccccctgca cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc
      841 gtacggggcg tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc
      901 cgggcggggc ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc
      961 ggagcgccgg cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt
     1021 gcgagagggc gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc
     1081 gccgccgcac cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa
     1141 tgggcgggga gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc
     1201 ggggctgtcc gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct
     1261 tctggcgtgt gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt
     1321 tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat
     1381 tcgccaccat ggaaggtttt gacgggtcag actttagccc tcctgccgat ctcgtgggtg
     1441 tggacggtgc cgtgatgcgg aacgtggtgg acgtgacaat caacggcgac gtgactgcac
     1501 cacctaaggc agctcctaga aaatctgaga gtgtgaagaa agtccactgg aacgacgtgg
     1561 atcagggacc aagcgagaag cccgaaacaa gacaggagaa gagcaggatc accagcgagg
     1621 gcgagtacat ccccctggac cagatcgaca tcaacgtgga cgacgggagg atggaaattc
     1681 ctaccacagg agtgggtgct agcatggcgg aaggatccgt cgccaggcag cctgacctct
     1741 tgacctgcga cgatgagccg atccatatcc ccggtgccat ccaaccgcat ggactgctgc
     1801 tcgccctcgc cgccgacatg acgatcgttg ccggcagcga caaccttccc gaactcaccg
     1861 gactggcgat cggcgccctg atcggccgct ctgcggccga tgtcttcgac tcggagacgc
     1921 acaaccgtct gacgatcgcc ttggccgagc ccggggcggc cgtcggagca ccgatcactg
     1981 tcggcttcac gatgcgaaag gacgcaggct tcatcggctc ctggcatcgc catgatcagc
     2041 tcatcttcct ggagctggag cctccccagc gggacgtcgc cgagccgcag gcgttcttcc
     2101 gccgcaccaa cagcgccatc cgccgcctgc aggccgccga aaccttggaa agcgcctgcg
     2161 ccgccgcggc gcaagaggtg cggaagatta ccggcttcga tcgggtgatg atctatcgct
     2221 tcgcctccga cttcagcggg tccgtgatcg cagaggatcg gtgcgccgag gtcgagtcaa
     2281 aactaggcct gcactatcct gcctcattca tcccggcgca ggcccgtcgg ctctatacca
     2341 tcaacccggt acggatcatt cccgacatca attatcggcc ggtgccggtc accccagacc
     2401 tcaatccggt caccgggcgg ccgattgatc ttagcttcgc catcctgcgc agcgtctcgc
     2461 ccaaccatct ggagttcatg cgcaacatag gcatgcacgg cacgatgtcg atctcgattt
     2521 tgcgcggcga gcgactgtgg ggattgatcg tttgccatca ccgaacgccg tactacgtcg
     2581 atctcgatgg ccgccaagcc tgcaagaggg tcgccgagag gctggccact cagatcggcg
     2641 tgatggaaga ggccgtagcc cgcgtccagt ttcgtgtccg agcagtgatt gatcatctag
     2701 ggatgcgagt ctttggagtc ttcctaattt tcttggacat catcctcatg atcattgatc
     2761 tcagtcttcc aggaaaaagt gaatcttcac aatcctttta tgacgggatg gctttggctc
     2821 tttcttgtta tttcatgctg gatttaggat taaggatatt tgcctacggg cccaagaatt
     2881 tcttcaccaa cccctgggag gttgctgatg gtttgattat cgttgtcaca ttcgtcgtca
     2941 cgatatttta cactgtgtta gatgaatacg tgcaagaaac aggagccgat ggtttggggc
     3001 agttggttgt gttggcccgt ttgctgcgta tcttcaagct gacccgccac ttcgtggccc
     3061 accaacaaat gaaggctgaa gccgcggcca aagaagccgc ggcaaaggag gcagcagcga
     3121 aagaagccgc ggccaaagcc cattcgaatt gcgaacatga agagatccac ctcgccggct
     3181 cgatccagcc gcatggcgcg cttctggtcg tcagcgaaca tgatcatcgc gtcatccagg
     3241 ccagcgccaa cgccgcggaa tttctgaatc tcggaagcgt actcggcgtt ccgctcgccg
     3301 agatcgacgg cgatctgttg atcaagatcc tgccgcatct cgatcccacc gccgaaggca
     3361 tgccggtcgc ggtgcgctgc cggatcggca atccctctac ggagtactgc ggtctgatgc
     3421 atcggcctcc ggaaggcggg ctgatcatcg aactcgaacg tgccggcccg tcgatcgatc
     3481 tgtcaggcac gctggcgccg gcgctggagc ggatccgcac ggcgggttca ctgcgcgcgc
     3541 tgtgcgatga caccgtgctg ctgtttcagc agtgcaccgg ctacgaccgg gtgatggtgt
     3601 atcgtttcga tgagcaaggc cacggcctgg tattctccga gtgccatgtg cctgggctcg
     3661 aatcctattt cggcaaccgc tatccgtcgt cgactgtccc gcagatggcg cggcagctgt
     3721 acgtgcggca gcgcgtccgc gtgctggtcg acgtcaccta tcagccggtg ccgctggagc
     3781 cgcggctgtc gccgctgacc gggcgcgatc tcgacatgtc gggctgcttc ctgcgctcga
     3841 tgtcgccgtg ccatctgcag ttcctgaagg acatgggcgt gcgcgccacc ctggcggtgt
     3901 cgctggtggt cggcggcaag ctgtggggcc tggttgtctg tcaccattat ctgccgcgct
     3961 tcatccgttt cgagctgcgg gcgatctgca aacggctcgc cgaaaggatc gcgacgcgga
     4021 tcaccgcgct tgagagctaa gcggccgcac tcctcaggtg caggctgcct atcagaaggt
     4081 ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc
     4141 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa
     4201 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     4261 tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata
     4321 tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa
     4381 acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt
     4441 ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt
     4501 tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct
     4561 cttatggaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc
     4621 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4681 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4741 ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac
     4801 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4861 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     4921 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4981 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     5041 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5101 tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5161 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5221 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5281 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5341 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5401 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5461 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5521 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5581 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5641 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5701 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5761 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5821 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5881 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5941 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6001 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6061 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6121 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6181 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6241 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6301 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6361 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6421 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6481 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6541 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6601 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6661 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6721 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6781 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6841 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6901 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6961 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7021 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7081 catttccccg aaaagtgcca cctgggtcga cattgattat tgactagtta ttaatagtaa
     7141 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     7201 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     7261 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     7321 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     7381 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     7441 tttcctactt ggca
//
