LOCUS       sequence-129558-        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-S127A mutant
FEATURES             Location/Qualifiers
     source          1..6231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        33..412
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        413..616
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     567..587
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             650..673
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      698..707
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             707..730
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2287..2909
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2486..2505)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2933..2953)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2938..3267
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3118..3253
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3305..3324)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3306..3324)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3338..3354)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3338..3354)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3351..3373)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3362..3378
                     /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(3386..3392,3393..3410,3411..3416))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3431..3452
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3569..3586)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3740..4328)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3820..3839)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4499..5290,5291..5359))
                     /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     5122..5141
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5360..5464)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5491..5946
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5578..5597)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5788..5809
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6072..6094
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6086..6103
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6087..6103
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta gttattaata
       61 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      121 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      181 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      241 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtccgccccc
      301 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttacg
      361 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      421 gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      481 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      541 atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt
      601 ctatataagc agagctcgtt tagtgaaccg tcagaattga tctaccatgg actacaaaga
      661 cgatgacgac aagcttgcgg ccgcgaattc aagcttagcc accatggact acaaagacga
      721 tgacgataaa gcaaggctcg aatcggtacc tatggatccc gggcagcagc cgccgcctca
      781 accggccccc cagggccaag ggcagccgcc ttcgcagccc ccgcaggggc agggcccgcc
      841 gtccggaccc gggcaaccgg cacccgcggc gacccaggcg gcgccgcagg caccccccgc
      901 cgggcatcag atcgtgcacg tccgcgggga ctcggagacc gacctggagg cgctcttcaa
      961 cgccgtcatg aaccccaaga cggccaacgt gccccagacc gtgcccatga ggctccggaa
     1021 gctgcccgac tccttcttca agccgccgga gcccaaatcc cactcccgac aggccagtac
     1081 tgatgcaggc actgcaggag ccctgactcc acagcatgtt cgagctcatg cctctccagc
     1141 ttctctgcag ttgggagctg tttctcctgg gacactgacc cccactggag tagtctctgg
     1201 cccagcagct acacccacag ctcagcatct tcgacagtct tcttttgaga tacctgatga
     1261 tgtacctctg ccagcaggtt gggagatggc aaagacatct tctggtcaga gatacttctt
     1321 aaatcacatc gatcagacaa caacatggca ggaccccagg aaggccatgc tgtcccagat
     1381 gaacgtcaca gcccccacca gtccaccagt gcagcagaat atgatgaact cggcttcagg
     1441 tcctcttcct gatggatggg aacaagccat gactcaggat ggagaaattt actatataaa
     1501 ccataagaac aagaccacct cttggctaga cccaaggctt gaccctcgtt ttgccatgaa
     1561 ccagagaatc agtcagagtg ctccagtgaa acagccacca cccctggctc cccagagccc
     1621 acagggaggc gtcatgggtg gcagcaactc caaccagcag caacagatgc gactgcagca
     1681 actgcagatg gagaaggaga ggctgcggct gaaacagcaa gaactgcttc ggcaggagtt
     1741 agccctgcgt agccagttac caacactgga gcaggatggt gggactcaaa atccagtgtc
     1801 ttctcccggg atgtctcagg aattgagaac aatgacgacc aatagctcag atcctttcct
     1861 taacagtggc acctatcact ctcgagatga gagtacagac agtggactaa gcatgagcag
     1921 ctacagtgtc cctcgaaccc cagatgactt cctgaacagt gtggatgaga tggatacagg
     1981 tgatactatc aaccaaagca ccctgccctc acagcagaac cgtttcccag actaccttga
     2041 agccattcct gggacaaatg tggaccttgg aacactggaa ggagatggaa tgaacataga
     2101 aggagaggag ctgatgccaa gtctgcagga agctttgagt tctgacatcc ttaatgacat
     2161 ggagtctgtt ttggctgcca ccaagctaga taaagaaagc tttcttacat ggttatagag
     2221 ccctcaggca gactgaattc tgcagatatc cagcacagtg gcggccgctc gagtctagag
     2281 gatcccgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc
     2341 actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag
     2401 gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg
     2461 aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa
     2521 tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc
     2581 gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag
     2641 agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac
     2701 ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc
     2761 cttctgattt taaaataact ataccagcag gaggacgtcc agacacagca taggctacct
     2821 ggccatgccc aaccggtggg acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt
     2881 gggtccactc agtagatgcc tgttgaattg ggtacgcggc cagcttggct gtggaatgtg
     2941 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3001 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3061 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3121 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3181 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3241 ttttttggag gcctaggctt ttgcaaaaag ctcctcgagg aactgaaaaa ccagaaagtt
     3301 aattccctat agtgagtcgt attaaattcg taatcatgtc atagctgttt cctgtgtgaa
     3361 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     3421 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     3481 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3541 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3601 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3661 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3721 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3781 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3841 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3901 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3961 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4021 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4081 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4141 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4201 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4261 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4321 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4381 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4441 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4501 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4561 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4621 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4681 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4741 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4801 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4861 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4921 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4981 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5041 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5101 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5161 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5221 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5281 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5341 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5401 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5461 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
     5521 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     5581 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     5641 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     5701 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     5761 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     5821 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     5881 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     5941 caatttccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     6001 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     6061 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tgatctatac
     6121 attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat aaatcaatat
     6181 tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt a
//
