LOCUS       sequence-271558-        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, Li L, Tumaneng K, Wang CY, Guan KL
  TITLE     A coordinated phosphorylation by Lats and CK1 regulates YAP 
            stability through SCF(beta-TRCP).
  JOURNAL   Genes Dev. 2010 Jan 1. 24(1):72-85.
  PUBMED    20048001
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. 
            2010 Jan 1. 24(1):72-85."
COMMENT     Sequence Label: pCMV-Flag YAP127/128/131/381A
FEATURES             Location/Qualifiers
     source          1..6231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             618..641
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      666..675
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             675..698
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2255..2877
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2454..2473)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2901..2921)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2906..3235
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3086..3221
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3273..3292)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3274..3292)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3306..3322)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3306..3322)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3319..3341)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3330..3346
                     /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(3354..3360,3361..3378,3379..3384))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3399..3420
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3537..3554)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3708..4296)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3788..3807)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4467..5258,5259..5327))
                     /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     5090..5109
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5328..5432)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5459..5914
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5546..5565)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5756..5777
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6040..6062
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6054..6071
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6055..6071
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattgatc taccatggac tacaaagacg atgacgacaa gcttgcggcc gcgaattcaa
      661 gcttagccac catggactac aaagacgatg acgataaagc aaggctcgaa tcggtaccta
      721 aggatcccgg gcagcagccg ccgcctcaac cggcccccca gggccaaggg cagccgcctt
      781 cgcagccccc gcaggggcag ggcccgccgt ccggacccgg gcaaccggca cccgcggcga
      841 cccaggcggc gccgcaggca ccccccgccg ggcatcagat cgtgcacgtc cgcggggact
      901 cggagaccga cctggaggcg ctcttcaacg ccgtcatgaa ccccaagacg gccaacgtgc
      961 cccagaccgt gcccatgagg ctccggaagc tgcccgactc cttcttcaag ccgccggagc
     1021 ccaaatccca ctcccgacag gccagtactg atgcaggcac tgcaggagcc ctgactccac
     1081 agcatgttcg agctcatgcc gctccagctg ctctgcagtt gggagctgtt tctcctggga
     1141 cactgacccc cactggagta gtctctggcc cagcagctac acccacagct cagcatcttc
     1201 gacagtcttc ttttgagata cctgatgatg tacctctgcc agcaggttgg gagatggcaa
     1261 agacatcttc tggtcagaga tacttcttaa atcacatcga tcagacaaca acatggcagg
     1321 accccaggaa ggccatgctg tcccagatga acgtcacagc ccccaccagt ccaccagtgc
     1381 agcagaatat gatgaactcg gcttcaggtc ctcttcctga tggatgggaa caagccatga
     1441 ctcaggatgg agaaatttac tatataaacc ataagaacaa gaccacctct tggctagacc
     1501 caaggcttga ccctcgtttt gccatgaacc agagaatcag tcagagtgct ccagtgaaac
     1561 agccaccacc cctggctccc cagagcccac agggaggcgt catgggtggc agcaactcca
     1621 accagcagca acagatgcga ctgcagcaac tgcagatgga gaaggagagg ctgcggctga
     1681 aacagcaaga actgcttcgg caggagttag ccctgcgtag ccagttacca acactggagc
     1741 aggatggtgg gactcaaaat ccagtgtctt ctcccgggat gtctcaggaa ttgagaacaa
     1801 tgacgaccaa tagctcagat cctttcctta acagtggcac ctatcactct cgagatgagg
     1861 ctacagacag tggactaagc atgagcagct acagtgtccc tcgaacccca gatgacttcc
     1921 tgaacagtgt ggatgagatg gatacaggtg atactatcaa ccaaagcacc ctgccctcac
     1981 agcagaaccg tttcccagac taccttgaag ccattcctgg gacaaatgtg gaccttggaa
     2041 cactggaagg agatggaatg aacatagaag gagaggagct gatgccaagt ctgcaggaag
     2101 ctttgagttc tgacatcctt aatgacatgg agtctgtttt ggctgccacc aagctagata
     2161 aagaaagctt tcttacatgg ttatagagcc ctcaggcaga ctgaattctg cagatatcca
     2221 gcacagtggc ggccgctcga gtctagagga tcccgggtgg catccctgtg acccctcccc
     2281 agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa
     2341 attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg
     2401 ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg
     2461 ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt
     2521 tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag
     2581 gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt
     2641 ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca
     2701 ggcgtgaacc actgctccct tccctgtcct tctgatttta aaataactat accagcagga
     2761 ggacgtccag acacagcata ggctacctgg ccatgcccaa ccggtgggac atttgagttg
     2821 cttgcttggc actgtcctct catgcgttgg gtccactcag tagatgcctg ttgaattggg
     2881 tacgcggcca gcttggctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     2941 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3001 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3061 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3121 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3181 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3241 cctcgaggaa ctgaaaaacc agaaagttaa ttccctatag tgagtcgtat taaattcgta
     3301 atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     3361 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     3421 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3481 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3541 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3601 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3661 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3721 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3781 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3841 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3901 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3961 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4021 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4081 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4141 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4201 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4261 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4321 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4381 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4441 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4501 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4561 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4621 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4681 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4741 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4801 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4861 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4921 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4981 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5041 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5101 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5161 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5221 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5281 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5341 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5401 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5461 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     5521 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     5581 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     5641 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5701 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5761 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5821 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5881 gcgaatttta acaaaatatt aacgcttaca atttccattc gccattcagg ctgcgcaact
     5941 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     6001 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     6061 cgacggccag tgccaagctg atctatacat tgaatcaata ttggcaatta gccatattag
     6121 tcattggtta tatagcataa atcaatattg gctattggcc attgcatacg ttgtatctat
     6181 atcataatat gtacatttat attggctcat gtccaatatg accgccatgt t
//
