LOCUS       sequence-151511-        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 WW mutant
FEATURES             Location/Qualifiers
     source          1..6231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1933..2388
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2020..2039)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2230..2251
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2514..2536
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2528..2545
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2529..2545
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        2706..3085
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3086..3289
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3240..3260
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3323..3346
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      3371..3380
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3380..3403
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    4960..5582
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5159..5178)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(5606..5626)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5611..5940
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5791..5926
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5978..5997)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5979..5997)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6011..6027)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6011..6027)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6024..6046)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6035..6051
                     /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(6059..6065,6066..6083,6084..6089))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6104..6125
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     1981 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2041 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2101 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2161 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2221 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2281 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     2341 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     2521 acgacgttgt aaaacgacgg ccagtgccaa gctgatctat acattgaatc aatattggca
     2581 attagccata ttagtcattg gttatatagc ataaatcaat attggctatt ggccattgca
     2641 tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc
     2701 atgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     2761 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     2821 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     2881 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     2941 acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc
     3001 cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta
     3061 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg
     3121 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     3181 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac
     3241 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
     3301 ccgtcagaat tgatctacca tggactacaa agacgatgac gacaagcttg cggccgcgaa
     3361 ttcaagctta gccaccatgg actacaaaga cgatgacgat aaagcaaggc tcgaatcggt
     3421 acctaaggat cccgggcagc agccgccgcc tcaaccggcc ccccagggcc aagggcagcc
     3481 gccttcgcag cccccgcagg ggcagggccc gccgtccgga cccgggcaac cggcacccgc
     3541 ggcgacccag gcggcgccgc aggcaccccc cgccgggcat cagatcgtgc acgtccgcgg
     3601 ggactcggag accgacctgg aggcgctctt caacgccgtc atgaacccca agacggccaa
     3661 cgtgccccag accgtgccca tgaggctccg gaagctgccc gactccttct tcaagccgcc
     3721 ggagcccaaa tcccactccc gacaggccag tactgatgca ggcactgcag gagccctgac
     3781 tccacagcat gttcgagctc attcctctcc agcttctctg cagttgggag ctgtttctcc
     3841 tgggacactg acccccactg gagtagtctc tggcccagca gctacaccca cagctcagca
     3901 tcttcgacag tcttcttttg agatacctga tgatgtacct ctgccagcag gttgggagat
     3961 ggcaaagaca tcttctggtc agagatactt cttaaatcac atcgatcaga caacaacagc
     4021 gcaggacgcc aggaaggcca tgctgtccca gatgaacgtc acagccccca ccagtccacc
     4081 agtgcagcag aatatgatga actcggcttc aggtcctctt cctgatggat gggaacaagc
     4141 catgactcag gatggagaaa tttactatat aaaccataag aacaagacca cctcttggct
     4201 agacccaagg cttgaccctc gttttgccat gaaccagaga atcagtcaga gtgctccagt
     4261 gaaacagcca ccacccctgg ctccccagag cccacaggga ggcgtcatgg gtggcagcaa
     4321 ctccaaccag cagcaacaga tgcgactgca gcaactgcag atggagaagg agaggctgcg
     4381 gctgaaacag caagaactgc ttcggcagga gttagccctg cgtagccagt taccaacact
     4441 ggagcaggat ggtgggactc aaaatccagt gtcttctccc gggatgtctc aggaattgag
     4501 aacaatgacg accaatagct cagatccttt ccttaacagt ggcacctatc actctcgaga
     4561 tgagagtaca gacagtggac taagcatgag cagctacagt gtccctcgaa ccccagatga
     4621 cttcctgaac agtgtggatg agatggatac aggtgatact atcaaccaaa gcaccctgcc
     4681 ctcacagcag aaccgtttcc cagactacct tgaagccatt cctgggacaa atgtggacct
     4741 tggaacactg gaaggagatg gaatgaacat agaaggagag gagctgatgc caagtctgca
     4801 ggaagctttg agttctgaca tccttaatga catggagtct gttttggctg ccaccaagct
     4861 agataaagaa agctttctta catggttata gagccctcag gcagactgaa ttctgcagat
     4921 atccagcaca gtggcggccg ctcgagtcta gaggatcccg ggtggcatcc ctgtgacccc
     4981 tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta
     5041 ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg
     5101 aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc
     5161 tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc
     5221 tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg
     5281 accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg
     5341 ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga
     5401 ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttaaaata actataccag
     5461 caggaggacg tccagacaca gcataggcta cctggccatg cccaaccggt gggacatttg
     5521 agttgcttgc ttggcactgt cctctcatgc gttgggtcca ctcagtagat gcctgttgaa
     5581 ttgggtacgc ggccagcttg gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     5641 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     5701 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5761 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     5821 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     5881 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     5941 aagctcctcg aggaactgaa aaaccagaaa gttaattccc tatagtgagt cgtattaaat
     6001 tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     6061 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     6121 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     6181 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc t
//
