LOCUS       sequence-193977-        7257 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Plasmid used (after linearization) to generate transgenic mice that 
            express the genetically encoded voltage indicator upon Cre 
            recombination..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7257)
  AUTHORS   Quinn TA, Camelliti P, Rog-Zielinska EA, Siedlecka U, Poggioli T, 
            O'Toole ET, Knopfel T, Kohl P
  TITLE     Electrotonic coupling of excitable and nonexcitable cells in the 
            heart revealed by optogenetics.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Dec 20;113(51):14852-14857. doi: 
            10.1073/pnas.1611184114. Epub 2016 Dec 7.
  PUBMED    27930302
REFERENCE   2  (bases 1 to 7257)
  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: 
            "10.1073/pnas.1611184114"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2016-12-20- 20"; volume: "113"; issue: "51"; pages: 
            "14852-14857"
COMMENT     Sequence Label: pCAG-VSFP2.3 rev. with Flex
FEATURES             Location/Qualifiers
     source          1..7257
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     enhancer        150..529
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        560..664
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(665..733,734..1525)
                     /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     complement(883..902)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1696..2284
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2185..2204
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2438..2455
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    2523..2657
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2547..2566)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2601..2620
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(2663..2858)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2677..2812
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    2917..2938
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2953..2958,2959..2976,2977..2983)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2991..3007
                     /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)."
     primer_bind     2996..3018
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3015..3031
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3015..3031
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3374..3393
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    3392..3447
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     3493..3512
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     misc_feature    3617..4205
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3723..3734
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4132..4152
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    4247..4280
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4351..4384
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     regulatory      4401..4410
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5169..5171,5172..5174,5175..5687)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIE"
     CDS             join(5865..5867,5868..5870,5871..6581)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     protein_bind    complement(6598..6631)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(6694..6727)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     complement(6900..6922)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
ORIGIN
        1 cgccccgccc cgccccgcct ggcgcgcgcc cccccccccc ccgcccccat cgctgcacaa
       61 aataattaaa aaataaataa atacaaaatt gggggtgggg agggggggga gatggggaga
      121 gtgaagcaga acgtggggct cacctcgacc atggtaatag cgatgactaa tacgtagatg
      181 tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat
      241 ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc
      301 aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc
      361 gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag
      421 ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact
      481 aatgaccccg taattgatta ctattaataa ctagtcaata atcaatgtcg acccaggtgg
      541 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
      601 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
      661 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
      721 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
      781 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
      841 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
      901 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
      961 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     1021 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     1081 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     1141 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     1201 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     1261 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     1321 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     1381 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     1441 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     1501 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     1561 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     1621 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     1681 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1741 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1801 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     1861 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1921 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     1981 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2041 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2101 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2161 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2221 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2281 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2341 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2401 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2461 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     2521 cggatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     2581 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     2641 gctgcaataa acaagttagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact
     2701 acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta
     2761 gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag
     2821 ttaggggcgg gactatggtt gctgactaat tgagatgcgg atccgctggc acgacaggtt
     2881 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     2941 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     3001 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg ggctgcaggt
     3061 cgagggatct ccataagaga agagggacag ctatgactgg gagtagtcag gagaggagga
     3121 aaaatctggc tagtaaaaca tgtaaggaaa attttaggga tgttaaagaa aaaaataaca
     3181 caaaacaaaa tataaaaaaa atctaacctc aagtcaaggc ttttctatgg aataaggaat
     3241 ggacagcagg gggctgtttc atatactgat gacctcttta tagccaacct ttgttcatgg
     3301 cagccagcat atgggcatat gttgccaaac tctaaaccaa atactcattc tgatgtttta
     3361 aatgatttgc cctcccatat gtccttccga gtgagagaca caaaaaattc caacacacta
     3421 ttgcaatgaa aataaatttc ctttattagc cagaagtcag atgctcaagg ggcttcatga
     3481 tgtccccata atttttggca gagggaaaaa gatctcagtg gtatttgtga gccagggcat
     3541 tggccacacc agccaccacc ttctgatagg cagcctgcac ctgaggagtg cggccgcctt
     3601 aagcgacggt atcgatgcgg ggaggcggcc caaagggaga tccgactcgt ctgagggcga
     3661 aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc cgcgggaagg aaggtccgct
     3721 ggattgaggg ccgaagggac gtagcagaag gacgtcccgc gcagaatcca ggtggcaaca
     3781 caggcgagca gccaaggaaa ggacgatgat ttccccgaca acaccacgga attgtcagtg
     3841 cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag ttccgccgtg
     3901 gcaataggga gggggaaagc gaaagtcccg gaaaggagct gacaggtggt ggcaatgccc
     3961 caaccagtgg gggttgcgtc agcaaacaca gtgcacacca cgccacgttg cctgacaacg
     4021 ggccacaact cctcataaag agacagcaac caggatttat acaaggagga gaaaatgaaa
     4081 gccatacggg aagcaatagc atgatacaaa ggcattaaag cagcgtatcc acatagcgta
     4141 aaaggagcaa catagttaag aataccagtc aatctttcac aaattttgta atccagaggt
     4201 tgattatcga taagcttgat atcgaattcg aacctcttcg agggacataa cttcgtataa
     4261 tgtatgctat acgaagttat gggtcgatgg tgagatctgg actagagggt cgatggtgat
     4321 gcttggatca aaataggaag aatcccaatg ataacttcgt ataaagtatc ctatacgaag
     4381 ttatactaga attcggtacc gccaccatgg agggattcga cggttcagat tttagtcctc
     4441 cagctgattt agttggcgtt gacggtgcag tcatgcggaa cgtcgttgac gtcacgataa
     4501 atggtgacgt cactgctccg ccgaaagctg cgccaagaaa aagtgaatcg gtaaagaaag
     4561 ttcattggaa tgatgtagac caaggaccga gtgaaaaacc agagacaaga caggaggaac
     4621 gaatagatat acccgagatt tcaggtctat ggtggggcga gaatgaacat ggagtggacg
     4681 atgggagaat ggagatacct actactggtg taggtcgcgt ccagtttcgt gtccgagcag
     4741 tgattgatca tctagggatg cgagtctttg gagtcttcct aattttcttg gacatcatcc
     4801 tcatgatcat tgatctcagt cttccaggaa aaagtgaatc ttcacaatcc ttttatgacg
     4861 ggatggcttt ggctctttct tgttatttca tgctggattt aggattaagg atatttgcct
     4921 acgggcccaa gaatttcttc accaacccct gggaggttgc tgatggtttg attatcgttg
     4981 tcacattcgt cgtcacgata ttttacactg tgttagatga atacgtgcaa gaaacaggag
     5041 ccgatggttt ggggcagttg gttgtgttgg cccgtttgct gcgtgtggtt cgattagcaa
     5101 gaatatttta ttcccaccaa caaatgaagg cttcaagcag aagaacaata tcacagaaca
     5161 agcgtcgaat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     5221 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     5281 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     5341 ggcccaccct cgtgaccacc ctgacctggg gcgtgcagtg cttcgcccgc taccccgacc
     5401 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     5461 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     5521 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     5581 tggggcacaa gctggagtac aacgccatca gcgacaacgt ctatatcacc gccgacaagc
     5641 agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     5701 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     5761 acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
     5821 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcatggtg agcaagggcg
     5881 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     5941 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     6001 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttcg
     6061 gctacggcct gatgtgcttc gcgcgctacc ccgaccacat gaagcagcac gacttcttca
     6121 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     6181 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     6241 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     6301 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     6361 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     6421 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
     6481 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     6541 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaggggat ccctagtata
     6601 acttcgtata gcatacatta tacgaagtta tgggtcgatg gtgatgcttg gcaattcggg
     6661 tcgatggtga agcattggtc ttcctatttt gatataactt cgtataggat actttatacg
     6721 aagttatggg gatccaattc tttgccaaaa tgatgagaca gcacaataac cagcacgttg
     6781 cccaggctag cgaattcttt gccaaaatga tgagacagca caataaccag cacgttgccc
     6841 aggagctgta ggaaaaagaa gaaggcatga acatggttag cagaggctct agagccgccg
     6901 gtcacacgcc agaagccgaa ccccgccctg ccccgtcccc cccgaaggca gccgtccccc
     6961 cgcggacagc cccgaggctg gagagggaga aggggacggc ggcgcggcga cgcacgaagg
     7021 ccctccccgc ccatttcctt cctgccggcg ccgcaccgct tcgccccgcg cccgctagag
     7081 ggggtgcggc ggcgcctccc agatttcggc tccgcacaga tttgggacaa aggaagtccc
     7141 tgcgccctct cgcacgatta ccataaaagg caatggctgc ggctcgccgc gcctcgacag
     7201 ccgccggcgc tccgggggcc gccgcgcccc tcccccgagc cctccccggc ccgaggc
//
