LOCUS       sequence-340494-        8116 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression of FLAG-tagged WWC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8116)
  AUTHORS   Ma X, Mandausch FJ, Wu Y, Sahoo VK, Ma W, Leoni G, Hostiuc M, 
            Wintgens JP, Qiu J, Kannaiyan N, Rossner MJ, Wehr MC
  TITLE     Comprehensive split TEV based protein-protein interaction screening 
            reveals TAOK2 as a key modulator of Hippo signalling to limit 
            growth.
  JOURNAL   Cell Signal. 2024 Jan;113:110917. doi: 
            10.1016/j.cellsig.2023.110917. Epub 2023 Oct 7.
  PUBMED    37813295
REFERENCE   2  (bases 1 to 8116)
  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: "Cell 
            Signal."; date: "2024-01"; volume: "113:110917. doi"; pages: " 
            10.1016/j.cellsig.2023.110917. Epub 2023 Oct 7"
COMMENT     Sequence Label: pCMV7.1_3xFLAG-WWC1
FEATURES             Location/Qualifiers
     source          1..8116
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             931..996
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    1033..1057
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4400..4424)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    4457..5079
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4656..4675)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(5103..5123)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5108..5437
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5288..5423
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5475..5494)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5476..5494)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5508..5524)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5508..5524)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5521..5543)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5532..5548
                     /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(5556..5562,5563..5580,5581..5586))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5601..5622
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5739..5756)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5910..6498)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5990..6009)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6669..7460,7461..7529))
                     /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     7292..7311
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7530..7634)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7661..8089
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7748..7767)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7958..7979
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagcttg cggccgcgaa ttcatcgata
     1021 gatctgatat caacaagttt gtacaaaaaa gcaggctctc cccggccgga gctgcccctg
     1081 ccggagggct gggaggaggc gcgcgacttc gacggcaagg tctactacat agaccacacg
     1141 aaccgcacca ccagctggat cgacccgcgg gacaggtaca ccaaaccgct cacctttgct
     1201 gactgcatta gtgatgagtt gccgctagga tgggaagagg catatgaccc acaggttgga
     1261 gattacttca tagaccacaa caccaaaacc actcagattg aggatcctcg agtacaatgg
     1321 cggcgggagc aggaacatat gctgaaggat tacctggtgg tggcccagga ggctctgagt
     1381 gcacaaaagg agatctacca ggtgaagcag cagcgcctgg agcttgcaca gcaggagtac
     1441 cagcaactgc atgccgtctg ggagcataag ctgggctccc aggtcagctt ggtctctggt
     1501 tcatcatcca gctccaagta tgaccctgag atcctgaaag ctgaaattgc cactgcaaaa
     1561 tcccgggtca acaagctgaa gagagagatg gttcacctcc agcacgagct gcagttcaaa
     1621 gagcgtggct ttcagaccct gaagaaaatc gataagaaaa tgtctgatgc tcagggcagc
     1681 tacaaactgg atgaagctca ggctgtcttg agagaaacaa aagccatcaa aaaggctatt
     1741 acctgtgggg aaaaggaaaa gcaagatctc attaagagcc ttgccatgtt gaaggacggc
     1801 ttccgcactg acagggggtc tcactcagac ctgtggtcca gcagcagctc tctggagagt
     1861 tcgagtttcc cgctaccgaa acagtacctg gatgtgagct cccagacaga catctcggga
     1921 agcttcggca tcaacagcaa caatcagttg gcagagaagg tcagattgcg ccttcgatat
     1981 gaagaggcta agagaaggat cgccaacctg aagatccagc tggccaagct tgacagtgag
     2041 gcctggcctg gggtgctgga ctcagagagg gaccggctga tccttatcaa cgagaaggag
     2101 gagctgctga aggagatgcg cttcatcagc ccccgcaagt ggacccaggg ggaggtggag
     2161 cagctggaga tggcccggaa gcggctggaa aaggacctgc aggcagcccg ggacacccag
     2221 agcaaggcgc tgacggagag gttaaagtta aacagtaaga ggaaccagct tgtgagagaa
     2281 ctggaggaag ccacccggca ggtggcaact ctgcactccc agctgaaaag tctctcaagc
     2341 agcatgcagt ccctgtcctc aggcagcagc cccggatccc tcacgtccag ccggggctcc
     2401 ctggttgcat ccagcctgga ctcctccact tcagccagct tcactgacct ctactatgac
     2461 ccctttgagc agctggactc agagctgcag agcaaggtgg agttcctgct cctggagggg
     2521 gccaccggct tccggccctc aggctgcatc accaccatcc acgaggatga ggtggccaag
     2581 acccagaagg cagagggagg tggccgcctg caggctctgc gttccctgtc tggcacccca
     2641 aagtccatga cctccctatc cccacgttcc tctctctcct ccccctcccc accctgttcc
     2701 cctctcatgg ctgaccccct cctggctggt gatgccttcc tcaactcctt ggagtttgaa
     2761 gacccggagc tgagtgccac tctttgtgaa ctgagccttg gtaacagcgc ccaggaaaga
     2821 taccggctgg aggaaccagg aacggagggc aagcagctgg gccaagctgt gaatacggcc
     2881 caggggtgtg gcctgaaagt ggcctgtgtc tcagccgccg tatcggacga gtcagtggct
     2941 ggagacagtg gtgtgtacga ggcttccgtg cagagactgg gtgcttcaga agctgctgca
     3001 tttgacagtg acgaatcgga agcagtgggt gcgacccgaa ttcagattgc cctgaagtat
     3061 gatgagaaga ataagcaatt tgcaatatta atcatccagc tgagtaacct ttctgctctg
     3121 ttgcagcaac aagaccagaa agtgaatatc cgcgtggctg tccttccttg ctctgaaagc
     3181 acaacctgcc tgttccggac ccggcctctg gacgcctcag acactctagt gttcaatgag
     3241 gtgttctggg tatccatgtc ctatccagcc cttcaccaga agaccttaag agtcgatgtc
     3301 tgtaccaccg acaggagcca tctggaagag tgcctgggag gcgcccagat cagcctggcg
     3361 gaggtctgcc ggtctgggga gaggtcgact cgctggtaca accttctcag ctacaaatac
     3421 ttgaagaaac agagcaggga gctcaagcca gtgggagtca tggcccctgc ctcagggcct
     3481 gccagcacgg acgctgtgtc tgctctgttg gaacagacag cagtggagct ggagaagagg
     3541 caggagggca ggagcagcac acagacactg gaagacagct ggaggtatga ggagaccagt
     3601 gagaatgagg cagtagccga ggaagaggag gaggaggtgg aggaggagga gggagaagag
     3661 gatgttttca ccgagaaagc ctcacctgat atggatgggt acccagcatt aaaggtggac
     3721 aaagagacca acacggagac cccggcccca tcccccacag tggtgcgacc taaggaccgg
     3781 agagtgggca ccccgtccca ggggccattt cttcgaggga gcaccatcat ccgctctaag
     3841 accttctccc caggacccca gagccagtac gtgtgccggc tgaatcggag tgatagtgac
     3901 agctccactc tgtccaaaaa gccacctttt gttcgaaact ccctggagcg acgcagcgtc
     3961 cggatgaagc ggccttcctc ggtcaagtcg ctgcgctccg agcgtctgat ccgtacctcg
     4021 ctggacctgg agttagacct gcaggcgaca agaacctggc acagccaact gacccaggag
     4081 atctcggtgc tgaaggagct caaggagcag ctggaacaag ccaagagcca cggggagaag
     4141 gagctgccac agtggttgcg tgaggacgag cgtttccgcc tgctgctgag gatgctggag
     4201 aagcggcaga tggaccgagc ggagcacaag ggtgagcttc agacagacaa gatgatgagg
     4261 gcagctgcca aggatgtgca caggctccga ggccagagct gtaaggaacc cccagaagtt
     4321 cagtctttca gggagaagat ggcatttttc acccggcctc ggatgaatat cccagctctc
     4381 tctgcagatg acgtctaaga cccagctttc ttgtacaaag tggttgattc ggtaccagtc
     4441 gactctagag gatcccgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
     4501 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
     4561 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
     4621 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
     4681 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
     4741 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt
     4801 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca
     4861 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc
     4921 cttccctgtc cttctgattt taaaataact ataccagcag gaggacgtcc agacacagca
     4981 taggctacct ggccatgccc aaccggtggg acatttgagt tgcttgcttg gcactgtcct
     5041 ctcatgcgtt gggtccactc agtagatgcc tgttgaattg ggtacgcggc cagcttggct
     5101 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5161 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     5221 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     5281 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     5341 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     5401 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcctcgagg aactgaaaaa
     5461 ccagaaagtt aattccctat agtgagtcgt attaaattcg taatcatgtc atagctgttt
     5521 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5581 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5641 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5701 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5761 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5821 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5881 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5941 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6001 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6061 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6121 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6181 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6241 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6301 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6361 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6421 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6481 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6541 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6601 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6661 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6721 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6781 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6841 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6901 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6961 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     7021 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7081 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7141 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7201 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7261 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7321 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7381 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7441 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7501 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7561 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7621 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca
     7681 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     7741 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     7801 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     7861 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     7921 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     7981 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     8041 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     8101 ttaacgctta caattt
//
