LOCUS       sequence-340445-        8434 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression of FLAG-tagged WWC3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8434)
  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 8434)
  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-WWC3
FEATURES             Location/Qualifiers
     source          1..8434
                     /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(4718..4742)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    4775..5397
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4974..4993)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(5421..5441)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5426..5755
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5606..5741
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5793..5812)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5794..5812)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5826..5842)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5826..5842)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5839..5861)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5850..5866
                     /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(5874..5880,5881..5898,5899..5904))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5919..5940
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6057..6074)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6228..6816)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6308..6327)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6987..7778,7779..7847))
                     /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     7610..7629
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7848..7952)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7979..8407
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8066..8085)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8276..8297
                     /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 ccatgccttg gctgagcggc
     1081 ggccggcggc gacggcgggg acagccgcgc gaggccccgc gggagccgcc gccgtccgcg
     1141 cagccccagc gggaaccgcc gccgccggcg ccccccgccg cggtccccac gcccccagct
     1201 ccttccgcgc cgccgccgcg ggcccgggag agcgccgagc tgccgctgcc cgccggctgg
     1261 gaggaggcgc gagactacga cggccgcgtc ttttacattg accacaacac gcgccagacg
     1321 tcgtggatcg acccccgcga ccggataaca aagccattga cctttgccga ttgtgttggg
     1381 gacgaacttc ctttaggatg ggaaaccgta tatgataaac aaattggagt ttattacatg
     1441 gaccacataa ataaacttac ccagattgag gatccaagag aacagtggag gcgagagcaa
     1501 gagcgcatgt tgaaggaata tttaattgta gctcaggagg ctctcaatgc caagaaagaa
     1561 atctaccaga ttaagcagca gcggttcgag ctggcccagg aggaatacca gcagctgcac
     1621 aaaatgtgtg aggatgacag ccgctcgtac gccagctcat tctctggata ttcaacaaat
     1681 acaaagtatg acccacacca aattaaagca gaaatagcaa gtcgtcggga taggctttcc
     1741 agattaaaac gagagctgac ccagatgaag caggaactgc agtacaaaga aaagggggtg
     1801 gagaccctgc aagagattga tcggaagatg tcaagtactc acaccagcta caaactggat
     1861 gaggcgcagg ctatcatgag tgagctccgg accatcaaga aggccatttg cacaggcgag
     1921 aaggagaggc gggacctgat gcatagcctg gccaagctga ccgatagctt caagaacagc
     1981 tgctccgtta ccgactctct agtggacttc cctcaccatg taggcgtgcc tggcgatgct
     2041 ggtgtacccc agcagttctg cgatgccggc tcccagactg atatcatcgg ggagtttgtc
     2101 tttgatgata aaacaagact tgtagaccga gtcagactta attggcaata tgaagaagcc
     2161 agaaagagag tcgccaacat ccagcagcag ctggcccggc ttgacaatga gtcctggcca
     2221 agcacggccg aggctgacag ggaccggctg cagctcatca aggagaagga ggccctgctg
     2281 caggagcttc agctcatcat tgcacagcgc cgctctgcgg gagatgtggc ccggctggag
     2341 gaggagcggg agcgcctaga ggaggagctg cggcgtgcac gggccacctc cgcacagggc
     2401 gccacggaga ggattctgct tcaggaaaaa aggaattgtc tacttatgca gctggaagaa
     2461 gccacccgct taacatccta cctccagtcc cagttaaaaa gcctgtgtgc cagcaccctg
     2521 acggtgtcct cggggagcag ccgcgggtcc ctagcctcca gccgtgggtc tctggcctcc
     2581 agccggggct ccctgagctc ggtcagcttc acggacatct acggcctccc gcagtacgag
     2641 aagcccgacg ctgagggcag ccagcttcta cgcttcgacc tcattccctt cgactctctg
     2701 gggcgagatg ctcccttttc agagccccca ggcccctcgg gcttccacaa gcagaggcgg
     2761 tccctggaca cgccccagtc cctggcatcg ctgtcctccc gctcctcgct gtcctcgctg
     2821 tcgcccccaa gctcgcccct ggacacgccc ttcctccctg cctcacggga ctcgccgctg
     2881 gcgcagctgg cggacagctg tgaggggcca ggcctgggcg ccctagacag actgcgggca
     2941 cacgcctcgg ctatggggga cgaagactta ccaggcatgg cggcccttca gccacacggg
     3001 gtccccgggg acggggaagg gccgcacgag cgaggacccc caccagccag cgctcccgtg
     3061 ggtggaacag tgactcttcg agaagacagt gccaagaggt tggagaggag ggcacgccgc
     3121 atctccgcat gtctgtcgga ttattcgcta gccagcgaca gtggggtgtt tgaacctcta
     3181 accaaaagga acgaagatgc cgaggagcct gcctacggag acacggccag taacggagat
     3241 ccccagatcc acgtgggact cctgcgcgac agtggcagcg agtgtctcct cgtgcacgtg
     3301 ctgcagctga agaacccggc ggggctggcg gtgaaggaag actgcaaagt ccacatccga
     3361 gtctatttgc ccccactgga ctcgggcacg cccaacactt actgctccaa ggctctggag
     3421 ttccaagtgc cgctggtgtt taacgaagtg ttcagaatcc ccgtgcattc cagcgcgttg
     3481 acactgaagt cacttcagtt atacgtgtgt tcagtgactc cgcagctgca ggaagaactg
     3541 ctgggcattg ctcagattaa cctggctgac tatgacagtt tgagtgagat gcagctgcgc
     3601 tggcattccg tgcaggtgtt caccagctct gaaccatcaa ggacgcggga ggctgggtgt
     3661 gcaggagaga gctccgcccg ggaccctgca cacaccatct ccatctccgg caagacggat
     3721 gcggtgacgg tgctcctggc cagaaccacg gcacagctgc aggcggtgga gagggaactg
     3781 gccgaggagc gggccaagct ggagtacacg gaggaggagg tcctggagat ggagcgcaag
     3841 gaggagcagg ccgaggccat atccgagcgg agttggcaag cggactcggt ggatagcggc
     3901 tgtagcaact gcacccagac cagccctccg tacccagagc cctgttgcat gggtatcgac
     3961 tccatcctgg gccacccatt tgctgctcag gcagggcctt acagccccga gaaatttcag
     4021 ccctcgcctc ttaaggttga taaggaaacc aacacggaag atctctttct ggaagaagca
     4081 gccagcctcg tgaaggagcg gcccagccgc cgggcccgag ggtcgccttt tgttcggagt
     4141 ggcacgattg tccgttccca gacattctcg cctggagcac gaagccagta tgtttgcaga
     4201 ctttatcgta gtgacagcga cagttcaacg ctgccccgga agtccccctt tgtccgaaat
     4261 actttggaaa gacgaaccct tcgctataag cagtcatgca ggtcttccct ggctgagctc
     4321 atggcccgca cctccctgga cttggagctg gatctccagg cgtcgagaac acggcagagg
     4381 cagctgaatg aggagctctg cgccctccgt gagctgcggc agcggttgga ggacgcccag
     4441 ctccgtggcc agactgacct cccaccctgg gtgcttcggg acgagcggct ccgtggcctg
     4501 ctgcgggagg ccgagcggca gacaagacag accaaacttg actaccgtca tgagcaggcg
     4561 gctgagaaga tgctgaagaa ggcctccaag gagatctacc agctgcgtgg gcagagccac
     4621 aaagagccca tccaagtgca gacctttagg gagaagatag cattcttcac aaggccaagg
     4681 atcaacatac ctcctctccc agccgacgac gtctgagacc cagctttctt gtacaaagtg
     4741 gttgattcgg taccagtcga ctctagagga tcccgggtgg catccctgtg acccctcccc
     4801 agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa
     4861 attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg
     4921 ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg
     4981 ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt
     5041 tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag
     5101 gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt
     5161 ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca
     5221 ggcgtgaacc actgctccct tccctgtcct tctgatttta aaataactat accagcagga
     5281 ggacgtccag acacagcata ggctacctgg ccatgcccaa ccggtgggac atttgagttg
     5341 cttgcttggc actgtcctct catgcgttgg gtccactcag tagatgcctg ttgaattggg
     5401 tacgcggcca gcttggctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     5461 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     5521 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     5581 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     5641 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     5701 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     5761 cctcgaggaa ctgaaaaacc agaaagttaa ttccctatag tgagtcgtat taaattcgta
     5821 atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5881 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5941 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     6001 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6061 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6121 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6181 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6241 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6301 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6361 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6421 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6481 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6541 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6601 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6661 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6721 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6781 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6841 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6901 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6961 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7021 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7081 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7141 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7201 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7261 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7321 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7381 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7441 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7501 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7561 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7621 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7681 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7741 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7801 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7861 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7921 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7981 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     8041 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     8101 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     8161 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     8221 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     8281 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     8341 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     8401 gcgaatttta acaaaatatt aacgcttaca attt
//
