LOCUS       sequence-350060-        6548 bp    DNA     circular SYN 14-APR-2025
DEFINITION  expresses optofYap in zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6548)
  AUTHORS   Toh PJY, Lai JKH, Hermann A, Destaing O, Sheetz MP, Sudol M, 
            Saunders TE
  TITLE     Optogenetic control of YAP cellular localisation and function.
  JOURNAL   EMBO Rep. 2022 Sep 5;23(9):e54401. doi: 10.15252/embr.202154401. 
            Epub 2022 Jul 25.
  PUBMED    35876586
REFERENCE   2  (bases 1 to 6548)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "EMBO Rep.";
            date: "2022-09-5"; volume: "23(9):e54401. doi"; pages: " 
            10.15252/embr.202154401. Epub 2022 Jul 25"
COMMENT     Sequence Label: pCS2-BiNLS-fYap
FEATURES             Location/Qualifiers
     source          1..6548
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        940..958
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     940..957
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1038..1742
                     /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(1180..1198)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1448..1467
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3505..3639
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3529..3548)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3583..3602
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3752..3772)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3752..3770)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3791..3807)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3791..3807)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3804..3826)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3815..3831
                     /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(3839..3845,3846..3863,3864..3869))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3884..3905
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4022..4039)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4193..4781)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4273..4292)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4952..5743,5744..5812))
                     /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     5575..5594
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5813..5917)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5943..6398)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6080..6101)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6292..6311
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(6469..6548,1..905)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 ggcgtatatg gactcgtgcc aattcaatat ggtggatctg gaccccagcc aattcaatat
       61 ggcggacttg gcaccatgcc aattcaatat ggcggacctg gcactgtgcc aactggggag
      121 gggtctactt ggcacggtgc caagtttgag gaggggtctt ggccctgtgc caagtccgcc
      181 atattgaatt ggcatggtgc caataatggc ggccatattg gctatatgcc aggatcaata
      241 tataggcaat atccaatatg gccctatgcc aatatggcta ttggccaggt tcaatactat
      301 gtattggccc tatgccatat agtattccat atatgggttt tcctattgac gtagatagcc
      361 cctcccaatg ggcggtccca tataccatat atggggcttc ctaataccgc ccatagccac
      421 tcccccattg acgtcaatgg tctctatata tggtctttcc tattgacgtc atatgggcgg
      481 tcctattgac gtatatggcg cctcccccat tgacgtcaat tacggtaaat ggcccgcctg
      541 gctcaatgcc cattgacgtc aataggacca cccaccattg acgtcaatgg gatggctcat
      601 tgcccattca tatccgttct cacgccccct attgacgtca atgacggtaa atggcccact
      661 tggcagtaca tcaatatcta ttaatagtaa cttggcaagt acattactat tggaagtacg
      721 ccagggtaca ttggcagtac tcccattgac gtcaatggcg gtaaatggcc cgcgatggct
      781 gccaagtaca tccccattga cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat
      841 tgacgtaaat gggcggtagg cgtgcctaat gggaggtcta tataagcaat gctcgtttag
      901 ggaaccgcca ttctgcctgg ggacgtcgga gcaagcttga tttaggtgac actatagaat
      961 acaagctact tgttcttttt gcaggatccc atcgattcga attcatgtta gccttgaaat
     1021 tagcaggtct tgatatcgtg agcaagggcg aggaggataa catggccatc atcaaggagt
     1081 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     1141 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     1201 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
     1261 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
     1321 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     1381 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     1441 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt
     1501 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     1561 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     1621 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     1681 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     1741 agggtggatc tggaggttca ggtggaagtt tggctactac acttgaacgt attgagaaga
     1801 actttgtcat tactgaccca agattgccag ataatcccat tatattcgcg tccgatagtt
     1861 tcttgcagtt gacagaatat agccgtgaag aaattttggg aagaaactgc aggtttctac
     1921 aaggtcctga aactgatcgc gcgacagtga gaaaaattag agatgccata gataaccaaa
     1981 cagaggtcac tgttcagctg attaattata caaagagtgg taaaaagttc tggaacctct
     2041 ttcacttgca gcctatgcga gatcagaagg gagatgtcca gtactttatt ggggttcagt
     2101 tggatggaac tgagcatgtc cgagatgctg ccgagagaga gggagtcatg ctgattaaga
     2161 aaactgcaga aaatattgac gaggccgcaa agagactgcc cgacgccaac ctggcagccg
     2221 cagccaagaa gaaaaagctg gacctcgaga tggatccgaa ccagcacaac cctccagccg
     2281 gccaccagat cgtccatgtt cggggagact ccgagaccga tctggaagct ctttttaacg
     2341 ctgtgatgaa cccgaaaaac accatcgtcc ccccttccgt gccgatgagg ttgagaaagc
     2401 tgccagactc attcttcacg ccgccagagc caaagtccca ctccagacaa gccagtacag
     2461 atgcaggtac tgctggtacc gtcacgcccc atcacgtccg ggcacactcc tcacctgcct
     2521 ccctgcagct gggcgcagtt tctcctggtg cactgaccag catgggtcca gcaaacgccc
     2581 cacctcagca cctccgccag tcctcttacg agatacccga tgacatgccg ctgcccccag
     2641 gctgggagat ggccaagacc ccttcgggac agagatactt cctaaaccat aatgaccaga
     2701 cgaccacctg gcaggaccca cgtaaagctc tccttcagat gaaccaggcc gccccagcca
     2761 gcccagtgcc tgtgcagcaa cagaacatca tgaacccagc ctcagcaatg aaccagcagc
     2821 gaatttccca gagtgcccca gtaaagcaag gctcccagtt gccctccagc ccacagagtg
     2881 gagtgatgag tggaaacaac cccatccgac tgcagcagat ccacatagag aaggagagac
     2941 tgaggatcaa acaagagctc cttcgccaga gacctcagga gctcgccctc agaaaccagc
     3001 tgccaaccag catggagcag gatggcggca cccaaaaccc tgtgtcttca cctggtatgg
     3061 gccaagacgc ccgtaacatg accaccaaca gctctgaccc tttcttgaac agcgggacat
     3121 atcactctag agacgagagc acagacagtg gcctgagcat gagcagttac agcgtccctc
     3181 gcactccaga cgacttcctc aacagtgtgg atgagatgga gacaggtgac accctcggcc
     3241 ccagctcaat ggcaacacag cccagccgat tccccgacta cctagacgcc atcccgggca
     3301 cggacgtgga cctaggcacc cttgagggcg agagcatggc ggtggagggc gaggagctga
     3361 tgcccagcct gcaggaagcc ctaagctccg acatcctcaa cgacatggag tcagtgctgg
     3421 cagctaccaa gatcgacaag gagaactttc taacctggct atagtagctc gagcctctag
     3481 aactatagtg agtcgtatta cgtagatcca gacatgataa gatacattga tgagtttgga
     3541 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     3601 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     3661 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aattcgcggc cgcggcgcca
     3721 atgcattggg cccggtaccc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     3781 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     3841 acatacgagc cgggagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     3901 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3961 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4021 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4081 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4141 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4201 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4261 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4321 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4381 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4441 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4501 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4561 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4621 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4681 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4741 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4801 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4861 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4921 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4981 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5041 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5101 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5161 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5221 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5281 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5341 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5401 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5461 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5521 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5581 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5641 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5701 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5761 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5821 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5881 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5941 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     6001 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     6061 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     6121 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     6181 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     6241 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     6301 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     6361 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
     6421 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagt cgaccgccaa
     6481 ttcaatatgg cgtatatgga ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa
     6541 ttcaatat
//
