LOCUS       sequence-350061-        8046 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses optoYAP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8046)
  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 8046)
  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"; journalName: "EMBO Rep.";
            date: "2022-09-5"; volume: "23(9):e54401. doi"; pages: " 
            10.15252/embr.202154401. Epub 2022 Jul 25"
COMMENT     Sequence Label: pDN34-BiNLS-hYAP1-1delta
FEATURES             Location/Qualifiers
     source          1..8046
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(115..134)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        306..685
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        686..889
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     840..860
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     934..953
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        934..952
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1017..1721
                     /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(1159..1177)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1427..1446
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     regulatory      2271..2280
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3708..3725)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3714..3938
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3984..4412
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4071..4090)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4281..4302
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4421..4441)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4426..4755
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4606..4741
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4668..4687
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4822..5616
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4876..4895)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5486..5505
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5790..5911
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5827..5846)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5881..5900
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5960..5976)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5960..5976)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5973..5995)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5984..6000
                     /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(6008..6014,6015..6032,6033..6038))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6053..6074
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6191..6208)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6362..6950)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6442..6461)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7121..7912,7913..7981))
                     /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     7744..7763
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7982..8046,1..40))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
       61 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
      121 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
      181 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
      241 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
      301 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
      361 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      421 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      481 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      541 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      601 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      661 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      721 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      781 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      841 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac
      901 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca
      961 agctggctag cgcagcgtct agaatgttag ccttgaaatt agcaggtctt gatatcgtga
     1021 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
     1081 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     1141 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
     1201 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
     1261 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     1321 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     1381 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
     1441 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     1501 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     1561 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     1621 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     1681 agggccgcca ctccaccggc ggcatggacg agctgtacaa gggtggatct ggaggttcag
     1741 gtggaagttt ggctactaca cttgaacgta ttgagaagaa ctttgtcatt actgacccaa
     1801 gattgccaga taatcccatt atattcgcgt ccgatagttt cttgcagttg acagaatata
     1861 gccgtgaaga aattttggga agaaactgca ggtttctaca aggtcctgaa actgatcgcg
     1921 cgacagtgag aaaaattaga gatgccatag ataaccaaac agaggtcact gttcagctga
     1981 ttaattatac aaagagtggt aaaaagttct ggaacctctt tcacttgcag cctatgcgag
     2041 atcagaaggg agatgtccag tactttattg gggttcagtt ggatggaact gagcatgtcc
     2101 gagatgctgc cgagagagag ggagtcatgc tgattaagaa aactgcagaa aatattgacg
     2161 aggccgcaaa gagactgccc gacgccaacc tggcagccgc agccaagaag aaaaagctgg
     2221 acggtaccgg aggggggggg gcccggaggc ggggagaggc gggggatccc gccaccatgg
     2281 atcccgggca gcagccgccg cctcaaccgg ccccccaggg ccaagggcag ccgccttcgc
     2341 agcccccgca ggggcagggc ccgccgtccg gacccgggca accggcaccc gcggcgaccc
     2401 aggcggcgcc gcaggcaccc cccgccgggc atcagatcgt gcacgtccgc ggggactcgg
     2461 agaccgacct ggaggcgctc ttcaacgccg tcatgaaccc caagacggcc aacgtgcccc
     2521 agaccgtgcc catgaggctc cggaagctgc ccgactcctt cttcaagccg ccggagccca
     2581 aatcccactc ccgacaggcc agtactgatg caggcactgc aggagccctg actccacagc
     2641 atgttcgagc tcattcctct ccagcttctc tgcagttggg agctgtttct cctgggacac
     2701 tgacccccac tggagtagtc tctggcccag cagctacacc cacagctcag catcttcgac
     2761 agtcttcttt tgagatacct gatgatgtac ctctgccagc aggttgggag atggcaaaga
     2821 catcttctgg tcagagatac ttcttaaatc acatcgatca gacaacaaca tggcaggacc
     2881 ccaggaaggc catgctgtcc cagatgaacg tcacagcccc caccagtcca ccagtgcagc
     2941 agaatatgat gaactcggct tcagccatga accagagaat cagtcagagt gctccagtga
     3001 aacagccacc acccctggct ccccagagcc cacagggagg cgtcatgggt ggcagcaact
     3061 ccaaccagca gcaacagatg cgactgcagc aactgcagat ggagaaggag aggctgcggc
     3121 tgaaacagca agaactgctt cggcaggtga ggccacaggc aatgcggaat atcaatccca
     3181 gcacagcaaa ttctccaaaa tgtcaggagt tagccctgcg tagccagtta ccaacactgg
     3241 agcaggatgg tgggactcaa aatccagtgt cttctcccgg gatgtctcag gaattgagaa
     3301 caatgacgac caatagctca gatcctttcc ttaacagtgg cacctatcac tctcgagatg
     3361 agagtacaga cagtggacta agcatgagca gctacagtgt ccctcgaacc ccagatgact
     3421 tcctgaacag tgtggatgag atggatacag gtgatactat caaccaaagc accctgccct
     3481 cacagcagaa ccgtttccca gactaccttg aagccattcc tgggacaaat gtggaccttg
     3541 gaacactgga aggagatgga atgaacatag aaggagagga gctgatgcca agtctgcagg
     3601 aagctttgag ttctgacatc cttaatgaca tggagtctgt tttggctgcc accaagctag
     3661 ataaagaaag ctttcttaca tggttatagg tttaaaccgc tgatcagcct cgactgtgcc
     3721 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3781 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3841 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3901 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3961 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4021 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4081 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4141 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4201 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4261 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4321 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4381 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     4441 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4501 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4561 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4621 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4681 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4741 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     4801 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4861 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4921 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4981 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     5041 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5101 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5161 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5221 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5281 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5341 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5401 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5461 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5521 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5581 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5641 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     5701 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5761 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     5821 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5881 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     5941 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6001 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6061 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6121 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6181 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6241 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6301 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6361 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6421 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6481 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6541 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6601 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6661 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6721 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6781 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6841 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6901 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6961 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7021 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7081 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7141 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7201 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7261 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7321 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7381 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7441 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7501 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7561 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7621 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7681 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7741 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7801 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7861 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7921 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7981 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8041 acatat
//
