LOCUS       sequence-345853-        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     complement(121..141)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        126..455
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      306..441
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(493..512)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(494..512)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(526..542)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(526..542)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(539..561)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    550..566
                     /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(574..580,581..598,599..604))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    619..640
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(757..774)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(928..1516)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1008..1027)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1687..2478,2479..2547))
                     /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     2310..2329
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2548..2652)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2679..3134
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2766..2785)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2976..2997
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3260..3282
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3274..3291
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3275..3291
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        3452..3831
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3832..4035
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3986..4006
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4065..4130
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    4167..4191
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6889..6913)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    join(6946..7471,1..97)
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(7145..7164)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
ORIGIN
        1 ggctacctgg ccatgcccaa ccggtgggac atttgagttg cttgcttggc actgtcctct
       61 catgcgttgg gtccactcag tagatgcctg ttgaattggg tacgcggcca gcttggctgt
      121 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
      181 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
      241 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
      301 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
      361 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
      421 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cctcgaggaa ctgaaaaacc
      481 agaaagttaa ttccctatag tgagtcgtat taaattcgta atcatgtcat agctgtttcc
      541 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
      601 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
      661 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
      721 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
      781 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
      841 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
      901 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
      961 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     1021 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     1081 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     1141 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     1201 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     1261 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     1321 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     1381 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     1441 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     1501 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     1561 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     1621 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     1681 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     1741 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     1801 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     1861 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     1921 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     1981 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     2041 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     2101 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     2161 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     2221 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     2281 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     2341 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     2401 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     2461 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     2521 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     2581 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     2641 aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt
     2701 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2761 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2821 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2881 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2941 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3001 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     3061 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     3121 aacgcttaca atttccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     3181 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     3241 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
     3301 atctatacat tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa
     3361 atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat
     3421 attggctcat gtccaatatg accgccatgt tgacattgat tattgactag ttattaatag
     3481 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     3541 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     3601 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     3661 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct
     3721 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg
     3781 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     3841 ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     3901 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     3961 tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     4021 tatataagca gagctcgttt agtgaaccgt cagaattaac catggactac aaagaccatg
     4081 acggtgatta taaagatcat gacatcgatt acaaggatga cgatgacaag cttgcggccg
     4141 cgaattcatc gatagatctg atatcaacaa gtttgtacaa aaaagcaggc tcccgtaaag
     4201 gggtgctgaa ggacccagag attgccgatc tattctacaa agatgatcct gaggaacttt
     4261 ttattggttt gcatgaaatt ggacatggaa gttttggagc agtttatttt gctacaaatg
     4321 ctcacaccag tgaggtggtg gcaattaaga agatgtccta tagtgggaag cagacccatg
     4381 agaaatggca agatattctt aaggaagtta aatttttacg acaattgaag catcctaata
     4441 ctattgagta caaaggctgt tacttgaaag aacacactgc ttggttggtg atggaatatt
     4501 gcttaggctc agcctctgat ttattagaag ttcataaaaa accacttcag gaagtggaga
     4561 tcgctgccat tactcatgga gccttgcatg gactagccta cctacattct catgcattga
     4621 ttcataggga tattaaagca ggaaatattc ttctaacaga gccaggtcag gtaaaactag
     4681 ctgattttgg atctgcttca atggcttctc ctgccaactc cttcgtgggc acaccttact
     4741 ggatggctcc agaggtgatc ttagctatgg atgaaggaca gtatgatggg aaagttgata
     4801 tttggtcact tggcatcact tgtattgaat tggcggaacg gaagccgccc cttttcaaca
     4861 tgaatgcaat gagtgcctta tatcacattg cccagaatga ctccccaacg ttacagtcta
     4921 atgaatggac agactccttt aggagatttg ttgattactg cttgcagaaa atacctcagg
     4981 aaaggccaac atcagcagaa ctattaaggc atgactttgt tcgacgagac cggccactac
     5041 gtgtcctcat tgacctcata cagaggacaa aagatgcagt tcgtgagcta gataacctac
     5101 agtaccgaaa aatgaaaaaa atacttttcc aagagacacg gaatggaccc ttgaatgagt
     5161 cacaggagga tgaggaagac agtgaacatg gaaccagcct gaacagggaa atggacagcc
     5221 tgggcagcaa ccattccatt ccaagcatgt ccgtgagcac aggcagccag agcagcagtg
     5281 tgaacagcat gcaggaagtc atggacgaga gcagttccga acttgtcatg atgcacgatg
     5341 acgaaagcac aatcaattcc agctcctccg tcgtgcataa gaaagatcat gtattcataa
     5401 gggatgaggc gggccacggc gatcccaggc ctgagccgcg gcctacccag tcagttcaga
     5461 gccaggccct ccactaccgg aacagagagc gctttgccac gatcaaatca gcatctttgg
     5521 ttacacgaca gatccatgag catgagcagg agaacgagtt gcgggaacag atgtcaggtt
     5581 ataagcggat gcggcgccag caccagaagc agctgatcgc cctggagaac aagctgaagg
     5641 ctgagatgga cgagcaccgc ctcaagctac agaaggaggt ggagacgcat gccaacaact
     5701 cgtccatcga gctggagaag ctggccaaga agcaagtggc tatcatagaa aaggaggcaa
     5761 aggtagctgc agcagatgag aagaagttcc agcaacagat cttggcccag cagaagaaag
     5821 atttgacaac tttcttagaa agtcagaaga agcagtataa gatttgtaag gaaaaaataa
     5881 aagaggaaat gaatgaggac catagcacac ccaagaaaga gaagcaagag cggatctcca
     5941 aacataaaga gaacttgcag cacacacagg ctgaagagga agcccacctt ctcactcaac
     6001 agagactgta ctacgacaaa aattgtcgtt tcttcaagcg gaaaataatg atcaagcggc
     6061 acgaggtgga gcagcagaac attcgggagg aactaaataa aaagaggacc cagaaggaga
     6121 tggagcatgc catgctaatc cggcacgacg agtccacccg agagctagag tacaggcagc
     6181 tgcacacgtt acagaagcta cgcatggatc tgatccgttt acagcaccag acggaactgg
     6241 aaaaccagct ggagtacaat aagaggcgag aaagagaact gcacagaaag catgtcatgg
     6301 aacttcggca acagccaaaa aacttaaagg ccatggaaat gcaaattaaa aaacagtttc
     6361 aggacacttg caaagtacag accaaacagt ataaagcact caagaatcac cagttggaag
     6421 ttactccaaa gaatgagcac aaaacaatct taaagacact gaaagatgag cagacaagaa
     6481 aacttgccat tttggcagag cagtatgaac agagtataaa tgaaatgatg gcctctcaag
     6541 cgttacggct agatgaggct caagaagcag aatgccaggc cttgaggcta cagctccagc
     6601 aggaaatgga gctgctcaac gcctaccaga gcaaaatcaa gatgcaaaca gaggcacaac
     6661 atgaacgtga gctccagaag ctagagcaga gagtgtctct gcgcagagca caccttgagc
     6721 agaagattga agaggagctg gctgcccttc agaaggaacg cagcgagaga ataaagaacc
     6781 tattggaaag gcaagagcga gagattgaaa cttttgacat ggagagcctc agaatgggat
     6841 ttgggaattt ggttacatta gattttccta aggaggacta cagatgagac ccagctttct
     6901 tgtacaaagt ggttgattcg gtaccagtcg actctagagg atcccgggtg gcatccctgt
     6961 gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt
     7021 gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta taatattatg
     7081 gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc
     7141 ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc
     7201 gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca
     7261 tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg
     7321 gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg
     7381 ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt aaaataacta
     7441 taccagcagg aggacgtcca gacacagcat a
//
