LOCUS       sequence-335607-        7471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression of FLAG-tagged TAOK3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7471)
  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 7471)
  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-TAOK3
FEATURES             Location/Qualifiers
     source          1..7471
                     /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(3755..3779)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3812..4434
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4011..4030)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4458..4478)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4463..4792
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4643..4778
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4830..4849)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4831..4849)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4863..4879)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4863..4879)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4876..4898)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4887..4903
                     /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(4911..4917,4918..4935,4936..4941))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4956..4977
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5094..5111)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5265..5853)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5345..5364)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6024..6815,6816..6884))
                     /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     6647..6666
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6885..6989)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7016..7444
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7103..7122)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7313..7334
                     /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 gcaggctccc gtaaaggggt gctgaaggac
     1081 ccagagattg ccgatctatt ctacaaagat gatcctgagg aactttttat tggtttgcat
     1141 gaaattggac atggaagttt tggagcagtt tattttgcta caaatgctca caccagtgag
     1201 gtggtggcaa ttaagaagat gtcctatagt gggaagcaga cccatgagaa atggcaagat
     1261 attcttaagg aagttaaatt tttacgacaa ttgaagcatc ctaatactat tgagtacaaa
     1321 ggctgttact tgaaagaaca cactgcttgg ttggtgatgg aatattgctt aggctcagcc
     1381 tctgatttat tagaagttca taaaaaacca cttcaggaag tggagatcgc tgccattact
     1441 catggagcct tgcatggact agcctaccta cattctcatg cattgattca tagggatatt
     1501 aaagcaggaa atattcttct aacagagcca ggtcaggtaa aactagctga ttttggatct
     1561 gcttcaatgg cttctcctgc caactccttc gtgggcacac cttactggat ggctccagag
     1621 gtgatcttag ctatggatga aggacagtat gatgggaaag ttgatatttg gtcacttggc
     1681 atcacttgta ttgaattggc ggaacggaag ccgccccttt tcaacatgaa tgcaatgagt
     1741 gccttatatc acattgccca gaatgactcc ccaacgttac agtctaatga atggacagac
     1801 tcctttagga gatttgttga ttactgcttg cagaaaatac ctcaggaaag gccaacatca
     1861 gcagaactat taaggcatga ctttgttcga cgagaccggc cactacgtgt cctcattgac
     1921 ctcatacaga ggacaaaaga tgcagttcgt gagctagata acctacagta ccgaaaaatg
     1981 aaaaaaatac ttttccaaga gacacggaat ggacccttga atgagtcaca ggaggatgag
     2041 gaagacagtg aacatggaac cagcctgaac agggaaatgg acagcctggg cagcaaccat
     2101 tccattccaa gcatgtccgt gagcacaggc agccagagca gcagtgtgaa cagcatgcag
     2161 gaagtcatgg acgagagcag ttccgaactt gtcatgatgc acgatgacga aagcacaatc
     2221 aattccagct cctccgtcgt gcataagaaa gatcatgtat tcataaggga tgaggcgggc
     2281 cacggcgatc ccaggcctga gccgcggcct acccagtcag ttcagagcca ggccctccac
     2341 taccggaaca gagagcgctt tgccacgatc aaatcagcat ctttggttac acgacagatc
     2401 catgagcatg agcaggagaa cgagttgcgg gaacagatgt caggttataa gcggatgcgg
     2461 cgccagcacc agaagcagct gatcgccctg gagaacaagc tgaaggctga gatggacgag
     2521 caccgcctca agctacagaa ggaggtggag acgcatgcca acaactcgtc catcgagctg
     2581 gagaagctgg ccaagaagca agtggctatc atagaaaagg aggcaaaggt agctgcagca
     2641 gatgagaaga agttccagca acagatcttg gcccagcaga agaaagattt gacaactttc
     2701 ttagaaagtc agaagaagca gtataagatt tgtaaggaaa aaataaaaga ggaaatgaat
     2761 gaggaccata gcacacccaa gaaagagaag caagagcgga tctccaaaca taaagagaac
     2821 ttgcagcaca cacaggctga agaggaagcc caccttctca ctcaacagag actgtactac
     2881 gacaaaaatt gtcgtttctt caagcggaaa ataatgatca agcggcacga ggtggagcag
     2941 cagaacattc gggaggaact aaataaaaag aggacccaga aggagatgga gcatgccatg
     3001 ctaatccggc acgacgagtc cacccgagag ctagagtaca ggcagctgca cacgttacag
     3061 aagctacgca tggatctgat ccgtttacag caccagacgg aactggaaaa ccagctggag
     3121 tacaataaga ggcgagaaag agaactgcac agaaagcatg tcatggaact tcggcaacag
     3181 ccaaaaaact taaaggccat ggaaatgcaa attaaaaaac agtttcagga cacttgcaaa
     3241 gtacagacca aacagtataa agcactcaag aatcaccagt tggaagttac tccaaagaat
     3301 gagcacaaaa caatcttaaa gacactgaaa gatgagcaga caagaaaact tgccattttg
     3361 gcagagcagt atgaacagag tataaatgaa atgatggcct ctcaagcgtt acggctagat
     3421 gaggctcaag aagcagaatg ccaggccttg aggctacagc tccagcagga aatggagctg
     3481 ctcaacgcct accagagcaa aatcaagatg caaacagagg cacaacatga acgtgagctc
     3541 cagaagctag agcagagagt gtctctgcgc agagcacacc ttgagcagaa gattgaagag
     3601 gagctggctg cccttcagaa ggaacgcagc gagagaataa agaacctatt ggaaaggcaa
     3661 gagcgagaga ttgaaacttt tgacatggag agcctcagaa tgggatttgg gaatttggtt
     3721 acattagatt ttcctaagga ggactacaga tgagacccag ctttcttgta caaagtggtt
     3781 gattcggtac cagtcgactc tagaggatcc cgggtggcat ccctgtgacc cctccccagt
     3841 gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt
     3901 aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt
     3961 ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga
     4021 accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca
     4081 agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct
     4141 cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc
     4201 caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc
     4261 gtgaaccact gctcccttcc ctgtccttct gattttaaaa taactatacc agcaggagga
     4321 cgtccagaca cagcataggc tacctggcca tgcccaaccg gtgggacatt tgagttgctt
     4381 gcttggcact gtcctctcat gcgttgggtc cactcagtag atgcctgttg aattgggtac
     4441 gcggccagct tggctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4501 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4561 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4621 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4681 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     4741 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctcct
     4801 cgaggaactg aaaaaccaga aagttaattc cctatagtga gtcgtattaa attcgtaatc
     4861 atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4921 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4981 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5041 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5101 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5161 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5221 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5281 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5341 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5401 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5461 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5521 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5581 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5641 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5701 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5761 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5821 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5881 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5941 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6001 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6061 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6121 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6181 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6241 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6301 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6361 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6421 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6481 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6541 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6601 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6661 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6721 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6781 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6841 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6901 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6961 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg
     7021 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     7081 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     7141 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     7201 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     7261 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     7321 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     7381 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     7441 aattttaaca aaatattaac gcttacaatt t
//
