LOCUS       sequence-259423-        6212 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Express FLAG-tagged YAP1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6212)
  AUTHORS   Bao Y, Nakagawa K, Yang Z, Ikeda M, Withanage K, Ishigami-Yuasa M, 
            Okuno Y, Hata S, Nishina H, Hata Y
  TITLE     A cell-based assay to screen stimulators of the Hippo pathway 
            reveals the inhibitory effect of dobutamine on the YAP-dependent 
            gene transcription.
  JOURNAL   J Biochem. 2011 Aug;150(2):199-208. doi: 10.1093/jb/mvr063. Epub 
            2011 May 17.
  PUBMED    21586534
REFERENCE   2  (bases 1 to 6212)
  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"; doi: "10.1093/jb/mvr063";
            journalName: "J Biochem"; date: "2011-08"; volume: "150"; issue: 
            "2"; pages: "199-208"
COMMENT     Sequence Label: pFLAG-YAP1
FEATURES             Location/Qualifiers
     source          1..6212
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             935..958
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2553..3175
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2752..2771)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3199..3219)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3204..3533
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3384..3519
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3571..3590)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3572..3590)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3604..3620)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3604..3620)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3617..3639)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3628..3644
                     /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(3652..3658,3659..3676,3677..3682))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3697..3718
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3835..3852)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4006..4594)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4086..4105)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4765..5556,5557..5625))
                     /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     5388..5407
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5626..5730)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5757..6185
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5844..5863)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6054..6075
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attgatctac catggactac aaagacgatg acgacaagct
      961 tgcggccgcg aattcatcga tagatcccgg gcagcagccg ccgcctcaac cggcccccca
     1021 gggccaaggg cagccgcctt cgcagccccc gcaggggcag ggcccgccgt ccggacccgg
     1081 gcaaccggca cccgcggcga cccaggcggc gccgcaggca ccccccgccg ggcatcagat
     1141 cgtgcacgtc cgcggggact cggagaccga cctggaggcg ctcttcaacg ccgtcatgaa
     1201 ccccaagacg gccaacgtgc cccagaccgt gcccatgagg ctccggaagc tgcccgactc
     1261 cttcttcaag ccgccggagc ccaaatccca ctcccgacag gccagtactg atgcaggcac
     1321 tgcaggagcc ctgactccac agcatgttcg agctcattcc tctccagctt ctctgcagtt
     1381 gggagctgtt tctcctggga cactgacccc cactggagta gtctctggcc cagcagctac
     1441 acccacagct cagcatcttc gacagtcttc ttttgagata cctgatgatg tacctctgcc
     1501 agcaggttgg gagatggcaa agacatcttc tggtcagaga tacttcttaa atcacatcga
     1561 tcagacaaca acatggcagg accccaggaa ggccatgctg tcccagatga acgtcacagc
     1621 ccccaccagt ccaccagtgc agcagaatat gatgaactcg gcttcaggtc ctcttcctga
     1681 tggatgggaa caagccatga ctcaggatgg agaaatttac tatataaacc ataagaacaa
     1741 gaccacctct tggctagacc caaggcttga ccctcgtttt gccatgaacc agagaatcag
     1801 tcagagtgct ccagtgaaac agccaccacc cctggctccc cagagcccac agggaggcgt
     1861 catgggtggc agcaactcca accagcagca acagatgcga ctgcagcaac tgcagatgga
     1921 gaaggagagg ctgcggctga aacagcaaga actgcttcgg caggcaatgc ggaatatcaa
     1981 tcccagcaca gcaaattctc caaaatgtca ggagttagcc ctgcgtagcc agttaccaac
     2041 actggagcag gatggtggga ctcaaaatcc agtgtcttct cccgggatgt ctcaggaatt
     2101 gagaacaatg acgaccaata gctcagatcc tttccttaac agtggcacct atcactctcg
     2161 agatgagagt acagacagtg gactaagcat gagcagctac agtgtccctc gaaccccaga
     2221 tgacttcctg aacagtgtgg atgagatgga tacaggtgat actatcaacc aaagcaccct
     2281 gccctcacag cagaaccgtt tcccagacta ccttgaagcc attcctggga caaatgtgga
     2341 ccttggaaca ctggaaggag atggaatgaa catagaagga gaggagctga tgccaagtct
     2401 gcaggaagct ttgagttctg acatccttaa tgacatggag tctgttttgg ctgccaccaa
     2461 gctagataaa gaaagctttc ttacatggtt atagagccct caggcagact gaattctgca
     2521 gtcgacggta ccagtcgact ctagaggatc ccgggtggca tccctgtgac ccctccccag
     2581 tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat
     2641 taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg
     2701 tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg
     2761 aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc
     2821 aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc
     2881 tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct
     2941 ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg
     3001 cgtgaaccac tgctcccttc cctgtccttc tgattttaaa ataactatac cagcaggagg
     3061 acgtccagac acagcatagg ctacctggcc atgcccaacc ggtgggacat ttgagttgct
     3121 tgcttggcac tgtcctctca tgcgttgggt ccactcagta gatgcctgtt gaattgggta
     3181 cgcggccagc ttggctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3241 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3301 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3361 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3421 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     3481 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3541 tcgaggaact gaaaaaccag aaagttaatt ccctatagtg agtcgtatta aattcgtaat
     3601 catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     3661 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     3721 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3781 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3841 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3901 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3961 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4021 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4081 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4141 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4201 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4261 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4321 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4381 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4441 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4501 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4561 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4621 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4681 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4741 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4801 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4861 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4921 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4981 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5041 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5101 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5161 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5221 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5281 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5341 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5401 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5461 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5521 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5581 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5641 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5701 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc
     5761 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5821 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5881 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     5941 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     6001 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     6061 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     6121 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     6181 gaattttaac aaaatattaa cgcttacaat tt
//
