LOCUS       sequence-220171-        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   Zhao B, Ye X, Yu J, Li L, Li W, Li S, Yu J, Lin JD, Wang CY, 
            Chinnaiyan AM, Lai ZC, Guan KL
  TITLE     TEAD mediates YAP-dependent gene induction and growth control.
  JOURNAL   Genes Dev. 2008 Jul 15. 22(14):1962-71.
  PUBMED    18579750
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"; journalName: "Genes Dev. 
            2008 Jul 15. 22(14):1962-71."
COMMENT     Sequence Label: pCMV-Flag-YAP-5SA/S94A
FEATURES             Location/Qualifiers
     source          1..6231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(88..107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     582..604
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     596..613
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     597..613
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        774..1153
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1154..1357
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1308..1328
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1391..1414
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      1439..1448
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1448..1471
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3028..3650
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3227..3246)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3674..3694)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3679..4008
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3859..3994
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4046..4065)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4047..4065)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4079..4095)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4079..4095)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4092..4114)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4103..4119
                     /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(4127..4133,4134..4151,4152..4157))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4172..4193
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4310..4327)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4481..5069)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4561..4580)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5240..6031,6032..6100))
                     /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     5863..5882
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6101..6205)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
       61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttccat tcgccattca ggctgcgcaa
      481 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
      541 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
      601 aacgacggcc agtgccaagc tgatctatac attgaatcaa tattggcaat tagccatatt
      661 agtcattggt tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct
      721 atatcataat atgtacattt atattggctc atgtccaata tgaccgccat gttgacatcg
      781 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      841 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      901 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      961 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     1021 tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     1081 tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat
     1141 cgctattacc atggtgatgc ggttttggca gtacaccaat gggcgtggat agcggtttga
     1201 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     1261 aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg
     1321 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagaattg
     1381 atctaccatg gactacaaag acgatgacga caagcttgcg gccgcgaatt caagcttagc
     1441 caccatggac tacaaagacg atgacgataa agcaaggctc gaatcggtac ctaaggatcc
     1501 cgggcagcag ccgccgcctc aaccggcccc ccagggccaa gggcagccgc cttcgcagcc
     1561 cccgcagggg cagggcccgc cgtccggacc cgggcaaccg gcacccgcgg cgacccaggc
     1621 ggcgccgcag gcaccccccg ccgggcatca gatcgtgcac gtccgcgggg acgcggagac
     1681 cgacctggag gcgctcttca acgccgtcat gaaccccaag acggccaacg tgccccagac
     1741 cgtgcccatg aggctccgga agctgcccga cgccttcttc aagccgccgg agcccaaatc
     1801 ccactcccga caggccgcta ctgatgcagg cactgcagga gccctgactc cacagcatgt
     1861 tcgagctcat gccgctccag ctgctctgca gttgggagct gtttctcctg ggacactgac
     1921 ccccactgga gtagtctctg gcccagcagc tacacccaca gctcagcatc ttcgacaggc
     1981 tgcttttgag atacctgatg atgtacctct gccagcaggt tgggagatgg caaagacatc
     2041 ttctggtcag agatacttct taaatcacat cgatcagaca acaacatggc aggaccccag
     2101 gaaggccatg ctgtcccaga tgaacgtcac agcccccacc agtccaccag tgcagcagaa
     2161 tatgatgaac tcggcttcag gtcctcttcc tgatggatgg gaacaagcca tgactcagga
     2221 tggagaaatt tactatataa accataagaa caagaccacc tcttggctag acccaaggct
     2281 tgaccctcgt tttgccatga accagagaat cagtcagagt gctccagtga aacagccacc
     2341 acccctggct ccccagagcc cacagggagg cgtcatgggt ggcagcaact ccaaccagca
     2401 gcaacagatg cgactgcagc aactgcagat ggagaaggag aggctgcggc tgaaacagca
     2461 agaactgctt cggcaggagt tagccctgcg tagccagtta ccaacactgg agcaggatgg
     2521 tgggactcaa aatccagtgt cttctcccgg gatgtctcag gaattgagaa caatgacgac
     2581 caatagctca gatcctttcc ttaacagtgg cacctatcac tctcgagatg aggctacaga
     2641 cagtggacta agcatgagca gctacagtgt ccctcgaacc ccagatgact tcctgaacag
     2701 tgtggatgag atggatacag gtgatactat caaccaaagc accctgccct cacagcagaa
     2761 ccgtttccca gactaccttg aagccattcc tgggacaaat gtggaccttg gaacactgga
     2821 aggagatgga atgaacatag aaggagagga gctgatgcca agtctgcagg aagctttgag
     2881 ttctgacatc cttaatgaca tggagtctgt tttggctgcc accaagctag ataaagaaag
     2941 ctttcttaca tggttataga gccctcaggc agactgaatt ctgcagatat ccagcacagt
     3001 ggcggccgct cgagtctaga ggatcccggg tggcatccct gtgacccctc cccagtgcct
     3061 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt
     3121 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta
     3181 tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca
     3241 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg
     3301 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc
     3361 taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac
     3421 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga
     3481 accactgctc ccttccctgt ccttctgatt ttaaaataac tataccagca ggaggacgtc
     3541 cagacacagc ataggctacc tggccatgcc caaccggtgg gacatttgag ttgcttgctt
     3601 ggcactgtcc tctcatgcgt tgggtccact cagtagatgc ctgttgaatt gggtacgcgg
     3661 ccagcttggc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3721 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3781 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3841 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3901 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3961 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcctcgag
     4021 gaactgaaaa accagaaagt taattcccta tagtgagtcg tattaaattc gtaatcatgt
     4081 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4141 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4201 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4261 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4321 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4381 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4441 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4501 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4561 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4621 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4681 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4741 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4801 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4861 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4921 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4981 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5041 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5101 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5161 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5221 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5281 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5341 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5401 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5461 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5521 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5581 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5641 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5701 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5761 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5821 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5881 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5941 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6001 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6061 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6121 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6181 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct g
//
