LOCUS       Exported                6863 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Genetically encoded fluorescent voltage indicator.
ACCESSION   .
VERSION     .
KEYWORDS    pCAG-Chimeric_Butterfly_YR_1.0
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6863)
  AUTHORS   Mishina Y, Mutoh H, Song C, Knopfel T
  TITLE     Exploration of genetically encoded voltage indicators based on a 
            chimeric voltage sensing domain.
  JOURNAL   Front Mol Neurosci. 2014 Sep 29;7:78. doi: 10.3389/fnmol.2014.00078.
            eCollection 2014.
  PUBMED    25324718
REFERENCE   2  (bases 1 to 6863)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6863
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1674
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1598..1620
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1682..1701
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3055..3774
                     /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(3100..3121)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3361..3380)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3708..3729
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3887..3906)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3952..4007
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4006..4025)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4366..4382)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4366..4382)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4379..4401)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4390..4406
                     /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(4414..4444)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4459..4480
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4539..4734
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4585..4720
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4647..4666
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    4740..4874
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4777..4796)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4831..4850
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4942..4959)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5113..5701)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5193..5212)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5872..6732)
                     /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     6495..6514
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6733..6837)
                     /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 ggctccgtac ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattcgc tagcgccacc
     1741 atgaggagcc gctttgtgaa gaaagatggt cattgcaatg ttcagtttat caacgtgatg
     1801 gtgtctaagg gcgaagagct gattaaggag aacatgcaca tgaagctgta catggagggc
     1861 accgtgaaca accaccactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc
     1921 acccagacca tgagaatcaa ggcggtcgag ggcggccctc tccccttcgc cttcgacatc
     1981 ctggctacca gcttcatgta cggcagcaaa accttcatca accacaccca gggcatcccc
     2041 gacttcttta agcagtcctt ccccgagggc ttcacatggg agagagtcac cacatacgaa
     2101 gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac
     2161 aacgtcaaga tcagaggggt gaacttccca tccaacggcc ctgtgatgca gaagaaaaca
     2221 ctcggctggg aggcctccac cgagaccctg taccccgctg acggcggcct ggaaggcaga
     2281 gccgacatgg ccctgaagct cgtgggcggg ggccacctga tctgcaactt gaagaccaca
     2341 tacagatcca agaaacccgc taagaacctc aagatgcccg gcgtctacta tgtggacaga
     2401 agactggaaa gaatcaagga ggccgacaaa gagacctacg tcgagcagca cgaggtggct
     2461 gtggccagat actgcgacct ccctagccga ctggggcaca gagaggaacg aatagatata
     2521 cccgagatat caggtctatg gtggggcgag aatgaacatg gagtggacga tgggagaatg
     2581 gagataccta ctactggtgt aggtcgcgtc cagtttcgtg tccgagcagt gattgatcat
     2641 ctagggatgc gagtctttgg agtcttccta attttcttgg acatcatcct catgatcatt
     2701 gatctcagtc ttccaggaaa aagtgaatct tcacaatcct tttatgacgg gatggctttg
     2761 gctctttctt gttatttcat gctggattta ggattaagga tatttgccta cgggcccaag
     2821 aatttcttca ccaacccctg ggaggttgct gatggtttga ttatcgttgt cacattcgtc
     2881 gtcacgatat tttacactgt gttagatgaa tacgtgcaag aaacaggagc cgatggtttg
     2941 gggcagttgg ttgtgttggc ccgtttgctg cgtatcttca agctgacccg ccacttcgtg
     3001 ggccaccaac aaatgaaggc ttcctcgaga agaacaatat cacagtgcgg ccgcatggtg
     3061 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     3121 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     3181 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     3241 accaccttcg gctacggcct gatgtgcttc gcgcgctacc ccgaccacat gaagcagcac
     3301 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     3361 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     3421 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     3481 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     3541 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     3601 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     3661 agctaccagt ccaaactgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     3721 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaggatcc
     3781 gagctcggta ccaagcttaa ggcggccgca ctcctcaggt gcaggctgcc tatcagaagg
     3841 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg
     3901 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     3961 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     4021 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     4081 atgccatatg ctggctgcca tgaacaaagg tggctataaa gaggtcatca gtatatgaaa
     4141 cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt
     4201 tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt
     4261 ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc
     4321 ttatgaagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc
     4381 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4441 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4501 cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc
     4561 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4621 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4681 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta
     4741 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4801 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4861 atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4921 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4981 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5041 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5101 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5161 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5221 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5281 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     5341 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5401 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5461 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5521 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     5581 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5641 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5701 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5761 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5821 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5881 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5941 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6001 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6061 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6121 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6181 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6241 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6301 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6361 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6421 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6481 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6541 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6601 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6661 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6721 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6781 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6841 atttccccga aaagtgccac ctg
//
