LOCUS       sequence-127553-        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-YAP2-1st&2nd WW mutant
FEATURES             Location/Qualifiers
     source          1..6231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     235..254
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        235..253
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(292..621)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      306..441
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     606..626
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    650..1272
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     1054..1073
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             complement(2829..2852)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      2852..2861
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(2886..2909)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2943..3146)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2972..2992)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3147..3526
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(3687..3704)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3687..3703)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3696..3718)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3844..4299)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3981..4002)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4193..4212
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4326..4430
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4431..4499,4500..5291)
                     /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     complement(4649..4668)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5462..6050
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5951..5970
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6204..6221
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aatttaatac
      241 gactcactat agggaattaa ctttctggtt tttcagttcc tcgaggagct ttttgcaaaa
      301 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggccgc
      361 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
      421 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
      481 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
      541 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
      601 tccacaccct aactgacaca cattccacag ccaagctggc cgcgtaccca attcaacagg
      661 catctactga gtggacccaa cgcatgagag gacagtgcca agcaagcaac tcaaatgtcc
      721 caccggttgg gcatggccag gtagcctatg ctgtgtctgg acgtcctcct gctggtatag
      781 ttattttaaa atcagaagga cagggaaggg agcagtggtt cacgcctgta atcccagcaa
      841 tttgggaggc caaggtgggt agatcacctg agattaggag ttggagacca gcctggccaa
      901 tatggtgaaa ccccgtctct accaaaaaaa caaaaattag ctgagcctgg tcatgcatgc
      961 ctggaatccc aacaactcgg gaggctgagg caggagaatc gcttgaaccc aggaggcgga
     1021 gattgcagtg agccaagatt gtgccactgc actccagctt ggttcccaat agaccccgca
     1081 ggccctacag gttgtcttcc caacttgccc cttgctccat accacccccc tccaccccat
     1141 aatattatag aaggacacct agtcagacaa aatgatgcaa cttaatttta ttaggacaag
     1201 gctggtgggc actggagtgg caacttccag ggccaggaga ggcactgggg aggggtcaca
     1261 gggatgccac ccgggatcct ctagactcga gcggccgcca ctgtgctgga tatctgcaga
     1321 attcagtctg cctgagggct ctataaccat gtaagaaagc tttctttatc tagcttggtg
     1381 gcagccaaaa cagactccat gtcattaagg atgtcagaac tcaaagcttc ctgcagactt
     1441 ggcatcagct cctctccttc tatgttcatt ccatctcctt ccagtgttcc aaggtccaca
     1501 tttgtcccag gaatggcttc aaggtagtct gggaaacggt tctgctgtga gggcagggtg
     1561 ctttggttga tagtatcacc tgtatccatc tcatccacac tgttcaggaa gtcatctggg
     1621 gttcgaggga cactgtagct gctcatgctt agtccactgt ctgtactctc atctcgagag
     1681 tgataggtgc cactgttaag gaaaggatct gagctattgg tcgtcattgt tctcaattcc
     1741 tgagacatcc cgggagaaga cactggattt tgagtcccac catcctgctc cagtgttggt
     1801 aactggctac gcagggctaa ctcctgccga agcagttctt gctgtttcag ccgcagcctc
     1861 tccttctcca tctgcagttg ctgcagtcgc atctgttgct gctggttgga gttgctgcca
     1921 cccatgacgc ctccctgtgg gctctgggga gccaggggtg gtggctgttt cactggagca
     1981 ctctgactga ttctctggtt catggcaaaa cgagggtcaa gccttgcgtc tagcgcagag
     2041 gtggtcttgt tcttatggtt tatatagtaa atttctccat cctgagtcat ggcttgttcc
     2101 catccatcag gaagaggacc tgaagccgag ttcatcatat tctgctgcac tggtggactg
     2161 gtgggggctg tgacgttcat ctgggacagc atggccttcc tggcgtcctg cgctgttgtt
     2221 gtctgatcga tgtgatttaa gaagtatctc tgaccagaag atgtctttgc catctcccaa
     2281 cctgctggca gaggtacatc atcaggtatc tcaaaagaag actgtcgaag atgctgagct
     2341 gtgggtgtag ctgctgggcc agagactact ccagtggggg tcagtgtccc aggagaaaca
     2401 gctcccaact gcagagaagc tggagaggaa tgagctcgaa catgctgtgg agtcagggct
     2461 cctgcagtgc ctgcatcagt actggcctgt cgggagtggg atttgggctc cggcggcttg
     2521 aagaaggagt cgggcagctt ccggagcctc atgggcacgg tctggggcac gttggccgtc
     2581 ttggggttca tgacggcgtt gaagagcgcc tccaggtcgg tctccgagtc cccgcggacg
     2641 tgcacgatct gatgcccggc ggggggtgcc tgcggcgccg cctgggtcgc cgcgggtgcc
     2701 ggttgcccgg gtccggacgg cgggccctgc ccctgcgggg gctgcgaagg cggctgccct
     2761 tggccctggg gggccggttg aggcggcggc tgctgcccgg gatccatagg taccgattcg
     2821 agccttgctt tatcgtcatc gtctttgtag tccatggtgg ctaagcttga attcgcggcc
     2881 gcaagcttgt cgtcatcgtc tttgtagtcc atggtagatc aattctgacg gttcactaaa
     2941 cgagctctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaacggg
     3001 gcggggttat tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca
     3061 ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca
     3121 ttggtgtact gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac
     3181 tgccaagtag gaaagtcccg taaggtcatg tactgggcat aatgccaggc gggccattta
     3241 ccgtcattga cgtcaatagg gggcggactt ggcatatgat acacttgatg tactgccaag
     3301 tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt
     3361 actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca
     3421 ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat
     3481 gaccccgtaa ttgattacta ttaataacta gtcaataatc aatgtcaaca tggcggtcat
     3541 attggacatg agccaatata aatgtacata ttatgatata gatacaacgt atgcaatggc
     3601 caatagccaa tattgattta tgctatataa ccaatgacta atatggctaa ttgccaatat
     3661 tgattcaatg tatagatcag cttggcactg gccgtcgttt tacaacgtcg tgactgggaa
     3721 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     3781 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     3841 tggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3901 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3961 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     4021 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     4081 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     4141 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     4201 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     4261 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     4321 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4381 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4441 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4501 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4561 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4621 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4681 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4741 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4801 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4861 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4921 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     4981 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5041 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5101 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5161 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5221 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5281 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5341 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5401 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5461 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5521 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5581 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5641 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5701 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5761 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5821 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5881 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5941 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6001 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6061 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6121 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     6181 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa g
//
