LOCUS       sequence-340440-        8359 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression of FLAG-tagged WWC2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8359)
  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 8359)
  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-WWC2
FEATURES             Location/Qualifiers
     source          1..8359
                     /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(4643..4667)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    4700..5322
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4899..4918)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(5346..5366)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5351..5680
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5531..5666
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5718..5737)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5719..5737)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5751..5767)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5751..5767)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5764..5786)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5775..5791
                     /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(5799..5805,5806..5823,5824..5829))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5844..5865
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5982..5999)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6153..6741)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6233..6252)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6912..7703,7704..7772))
                     /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     7535..7554
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7773..7877)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7904..8332
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7991..8010)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8201..8222
                     /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 gcaggctcca ccatgcctag gagggccggg
     1081 agcggtcagc tgccgctgcc ccggggctgg gaggaggcca gggactacga cggcaaggtc
     1141 ttctacattg accacaacac caggaggacc agctggatcg acccccggga caggttaacg
     1201 aagcccttgt catttgctga ttgtgttggg gatgagctgc cgtggggatg ggaagcaggg
     1261 tttgaccctc agattggtgt ctactacatc gatcacatca acaaaaccac gcagatagaa
     1321 gatccaagaa aacaatggag gggggaacag gagaagatgc tcaaggacta cctctctgtg
     1381 gcacaggatg ccctccggac acagaaggaa ctgtaccatg tgaaggagca gaggctggcg
     1441 ctggccctgg atgaatacgt gcgattaaat gatgcctata aggaaaagtc aagttctcac
     1501 acaagcttat tctcaggatc ttcatccagt actaaatatg atcccgatat tttaaaagct
     1561 gagatctcca ctacaagatt aagggttaaa aagctaaaga gagagctctc acagatgaag
     1621 caggaactgc tctataaaga acaaggcttt gaaacattgc agcaaattga taaaaaaatg
     1681 tctggaggcc agagcgggta tgaactcagt gaagccaaag ccattctaac agaactaaaa
     1741 tctatcagaa aggcaattag ctcaggagaa aaagaaaaac aagatctgat gcagagtctt
     1801 gctaagctgc aggagcggtt tcatttggat cagaacattg gcagatctga gccagatttg
     1861 agatgtagtc ctgtgaactc tcatttatgt ctctccagac agacccttga tgctgggtca
     1921 caaacaagca tttccggaga tattggagta agaagtagat caaatttagc tgaaaaggtc
     1981 aggctaagcc tacagtatga agaagccaaa agaagtatgg ccaacttaaa aattgaactg
     2041 tcaaaattgg acagtgaggc ctggcctggg gcactggata ttgagaagga aaaactgatg
     2101 ctgattaatg aaaaagaaga acttttgaaa gagcttcagt tcgtcacccc acagaaacgt
     2161 acccaagatg aattagaacg cctagaagct gaaaggcagc ggctggaaga agagttgctg
     2221 tctgtgaggg gaacaccaag cagagctctg gccgagagat tgagattgga agagagaaga
     2281 aaagagctgc tacagaaact tgaagaaact actaaattaa ctacttattt gcattcacaa
     2341 cttaaaagcc tctctgccag caccctgtcc atgtcatctg ggagcagcct gggttccctg
     2401 gcatcgagtc ggggctctct gaacacctcc agcagagggt cactcaactc cctcagttcc
     2461 accgaactct attacagcag tcaaagtgat cagatagatg tggattatca gtataaactg
     2521 gacttccttc tgcaagagaa aagcggttac attccttctg gacccatcac caccatccat
     2581 gaaaacgagg tggtcaagtc ccctagccag cctggccaga gtggactctg tggagtggca
     2641 gctgcagcaa caggccacac tcctccactg gctgaggccc cgaagtctgt ggcctccctg
     2701 tcctcgaggt cctccctttc ctccttgtct cctccaggct ctcccttggt tttggaaggc
     2761 acgtttccca tgtcttcttc tcatgatgcc tctctccatc agttcactgc tgactttgaa
     2821 gactgtgagt tgagtagcca ttttgcagat atcagcctca tcgaaaatca gattttgctg
     2881 gattctgatt caggaggagc ctcccagtct ctttcagagg ataaagacct taatgaatgt
     2941 gctagggagc cattatatga aggaactgca gatgtggaaa aatcattacc aaaaagaaga
     3001 gtgatccact tgcttgggga gaaaaccact tgtgtgtcgg ctgctgtgtc tgatgagtct
     3061 gtggctggag acagtggggt ctatgaagct ttcgtgaaac aacctagtga aatggaagat
     3121 gtcacataca gtgaagagga tgtagccatt gtagagaccg cccaggttca gataggactc
     3181 agatacaatg caaaaagttc aagtttcatg gtgattatag cacagctccg aaaccttcat
     3241 gccttcttga tacctcatac ttcaaaagta tattttaggg ttgccgttct tccttcctca
     3301 actgatgtca gctgtctgtt tcgcacaaaa gttcatccgc ccacagaatc cattttattc
     3361 aatgatgtgt tcagagtcgc catttcccaa acagccttac aacagaagac actgagggta
     3421 gacctttgct ctgtcagtaa acaccgaagg gaagaatgcc tggctggaac tcagatcagc
     3481 ctggcagatt taccattttc cagtgaggtt ttcactctat ggtataactt gcttccttcc
     3541 aagcaaatgc cttgcaaaaa gaatgaagaa aatgaggact ctgtatttca accaaaccag
     3601 ccgttagtag attctataga cttggatgca gtgtcagcct tacttgcaag aacatcagct
     3661 gagttgttag ctgtggaaca agaattagca caagaagaag aagaagaatc aggacaagaa
     3721 gagccaaggg gcccagatgg agactggcta acaatgctaa gagaggcctc tgatgaaatt
     3781 gtggctgaaa aagaggctga agttaaattg ccagaggaca gtagctgtac agaagattta
     3841 agttcatgca ctagtgtgcc tgagatgaat gaagacggga acaggaaaga aagcaactgt
     3901 gccaaagacc tcagaagtca gccacctact agaataccaa cactggttga caaagagaca
     3961 aacactgatg aagccgctaa tgacaatatg gcagttcgcc ccaaagagcg cagcagcctg
     4021 agctctagac agcatccgtt tgtgaggagc agtgtgatag tgcgctcaca gaccttttct
     4081 ccaggagagc ggaaccagta catctgcagg ttaaatcgga gtgacagtga cagttcaacc
     4141 ctggctaaaa aatcactgtt tgtgagaaac tccaccgaac gccgcagttt gagggtcaaa
     4201 aggacggttt gccagtcagt ccttagaaga acaacacagg aatgcccagt gcggacatct
     4261 ctagacttag aactggacct tcaggcatct ctgacccggc agagccgcct caatgatgag
     4321 ctgcaggcgc tgagggactt gcggcagaag ctggaggaac tgaaagctca gggagagact
     4381 gaccttccac caggcgtgct ggaggatgag aggttccaga ggcttctgaa gcaagctgag
     4441 aagcaggctg aacagtccaa agaagagcag aagcaaggtc tgaatgcaga gaagttgatg
     4501 aggcaagtct ccaaggacgt gtgtcggctc cgggagcaga gccagaaggt gcctcggcag
     4561 gtgcagtcct tcagggagaa gattgcctac ttcaccagag caaagataag catcccatcc
     4621 ctgccagctg atgatgtgtg agacccagct ttcttgtaca aagtggttga ttcggtacca
     4681 gtcgactcta gaggatcccg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
     4741 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
     4801 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
     4861 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
     4921 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
     4981 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
     5041 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc
     5101 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
     5161 tcccttccct gtccttctga ttttaaaata actataccag caggaggacg tccagacaca
     5221 gcataggcta cctggccatg cccaaccggt gggacatttg agttgcttgc ttggcactgt
     5281 cctctcatgc gttgggtcca ctcagtagat gcctgttgaa ttgggtacgc ggccagcttg
     5341 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     5401 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     5461 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     5521 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     5581 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     5641 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcctcg aggaactgaa
     5701 aaaccagaaa gttaattccc tatagtgagt cgtattaaat tcgtaatcat gtcatagctg
     5761 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5821 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5881 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5941 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     6001 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6061 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6121 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6181 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6241 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6301 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6361 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6421 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6481 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6541 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6601 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6661 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6721 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6781 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6841 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6901 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6961 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7021 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7081 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7141 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7201 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7261 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7321 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7381 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7441 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7501 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7561 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7621 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7681 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7741 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7801 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7861 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc
     7921 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     7981 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     8041 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     8101 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     8161 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     8221 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     8281 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     8341 atattaacgc ttacaattt
//
