LOCUS       sequence-129561-        6231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6231)
  AUTHORS   Oka T, Mazack V, Sudol M
  TITLE     Mst2 and Lats kinases regulate apoptotic function of Yes 
            kinase-associated protein (YAP).
  JOURNAL   J Biol Chem. 2008 Oct 10;283(41):27534-27546. doi: 
            10.1074/jbc.M804380200. Epub 2008 Jul 17.
  PUBMED    18640976
REFERENCE   2  (bases 1 to 6231)
  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: 
            "10.1074/jbc.M804380200"; journalName: "J Biol Chem"; date: 
            "2008-10-10- 10"; volume: "283"; issue: "41"; pages: "27534-27546"
COMMENT     Sequence Label: p2xFlag CMV2-YAP2-S127D mutant
FEATURES             Location/Qualifiers
     source          1..6231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        32..411
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        412..615
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     566..586
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             649..672
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      697..706
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             706..729
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2286..2908
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2485..2504)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2932..2952)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2937..3266
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3117..3252
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3304..3323)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3305..3323)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3337..3353)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3337..3353)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3350..3372)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3361..3377
                     /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(3385..3391,3392..3409,3410..3415))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3430..3451
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3568..3585)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3739..4327)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3819..3838)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4498..5289,5290..5358))
                     /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     5121..5140
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5359..5463)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5490..5945
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5577..5596)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5787..5808
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6071..6093
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6085..6102
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6086..6102
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 attggctcat gtccaatatg accgccatgt tgacattgat tattgactag ttattaatag
       61 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      121 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      181 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      241 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct
      301 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg
      361 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      421 ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      481 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      541 tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      601 tatataagca gagctcgttt agtgaaccgt cagaattgat ctaccatgga ctacaaagac
      661 gatgacgaca agcttgcggc cgcgaattca agcttagcca ccatggacta caaagacgat
      721 gacgataaag caaggctcga atcggtacct atggatcccg ggcagcagcc gccgcctcaa
      781 ccggcccccc agggccaagg gcagccgcct tcgcagcccc cgcaggggca gggcccgccg
      841 tccggacccg ggcaaccggc acccgcggcg acccaggcgg cgccgcaggc accccccgcc
      901 gggcatcaga tcgtgcacgt ccgcggggac tcggagaccg acctggaggc gctcttcaac
      961 gccgtcatga accccaagac ggccaacgtg ccccagaccg tgcccatgag gctccggaag
     1021 ctgcccgact ccttcttcaa gccgccggag cccaaatccc actcccgaca ggccagtact
     1081 gatgcaggca ctgcaggagc cctgactcca cagcatgttc gagctcatga ctctccagct
     1141 tctctgcagt tgggagctgt ttctcctggg acactgaccc ccactggagt agtctctggc
     1201 ccagcagcta cacccacagc tcagcatctt cgacagtctt cttttgagat acctgatgat
     1261 gtacctctgc cagcaggttg ggagatggca aagacatctt ctggtcagag atacttctta
     1321 aatcacatcg atcagacaac aacatggcag gaccccagga aggccatgct gtcccagatg
     1381 aacgtcacag cccccaccag tccaccagtg cagcagaata tgatgaactc ggcttcaggt
     1441 cctcttcctg atggatggga acaagccatg actcaggatg gagaaattta ctatataaac
     1501 cataagaaca agaccacctc ttggctagac ccaaggcttg accctcgttt tgccatgaac
     1561 cagagaatca gtcagagtgc tccagtgaaa cagccaccac ccctggctcc ccagagccca
     1621 cagggaggcg tcatgggtgg cagcaactcc aaccagcagc aacagatgcg actgcagcaa
     1681 ctgcagatgg agaaggagag gctgcggctg aaacagcaag aactgcttcg gcaggagtta
     1741 gccctgcgta gccagttacc aacactggag caggatggtg ggactcaaaa tccagtgtct
     1801 tctcccggga tgtctcagga attgagaaca atgacgacca atagctcaga tcctttcctt
     1861 aacagtggca cctatcactc tcgagatgag agtacagaca gtggactaag catgagcagc
     1921 tacagtgtcc ctcgaacccc agatgacttc ctgaacagtg tggatgagat ggatacaggt
     1981 gatactatca accaaagcac cctgccctca cagcagaacc gtttcccaga ctaccttgaa
     2041 gccattcctg ggacaaatgt ggaccttgga acactggaag gagatggaat gaacatagaa
     2101 ggagaggagc tgatgccaag tctgcaggaa gctttgagtt ctgacatcct taatgacatg
     2161 gagtctgttt tggctgccac caagctagat aaagaaagct ttcttacatg gttatagagc
     2221 cctcaggcag actgaattct gcagatatcc agcacagtgg cggccgctcg agtctagagg
     2281 atcccgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca
     2341 ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg
     2401 tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga
     2461 agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat
     2521 cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg
     2581 agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga
     2641 gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc
     2701 caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc
     2761 ttctgatttt aaaataacta taccagcagg aggacgtcca gacacagcat aggctacctg
     2821 gccatgccca accggtggga catttgagtt gcttgcttgg cactgtcctc tcatgcgttg
     2881 ggtccactca gtagatgcct gttgaattgg gtacgcggcc agcttggctg tggaatgtgt
     2941 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3001 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3061 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3121 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3181 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     3241 tttttggagg cctaggcttt tgcaaaaagc tcctcgagga actgaaaaac cagaaagtta
     3301 attccctata gtgagtcgta ttaaattcgt aatcatgtca tagctgtttc ctgtgtgaaa
     3361 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3421 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3481 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3541 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3601 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3661 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3721 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3781 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3841 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3901 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3961 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4021 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4081 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4141 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4201 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4261 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4321 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4381 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4441 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4501 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4561 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4621 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4681 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4741 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4801 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4861 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4921 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4981 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5041 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5101 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5161 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5221 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5281 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5341 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5401 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5461 gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc
     5521 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     5581 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     5641 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     5701 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     5761 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     5821 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     5881 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     5941 aatttccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     6001 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     6061 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct gatctataca
     6121 ttgaatcaat attggcaatt agccatatta gtcattggtt atatagcata aatcaatatt
     6181 ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta t
//
