LOCUS       sequence-340254-        8482 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression of FLAG-tagged TAOK2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8482)
  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 8482)
  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-TAOK2
FEATURES             Location/Qualifiers
     source          1..8482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             614..679
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    716..740
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4449..4473)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    4506..5128
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4705..4724)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(5152..5172)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5157..5486
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5337..5472
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5524..5543)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5525..5543)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5557..5573)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5557..5573)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5570..5592)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5581..5597
                     /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(5605..5611,5612..5629,5630..5635))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5650..5671
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5788..5805)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5959..6547)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6039..6058)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6718..7509,7510..7578))
                     /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     7341..7360
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7579..7683)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7710..8165
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7797..7816)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8007..8028
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8291..8313
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8305..8322
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8306..8322
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattaacc atggactaca aagaccatga cggtgattat aaagatcatg acatcgatta
      661 caaggatgac gatgacaagc ttgcggccgc gaattcatcg atagatctga tatcaacaag
      721 tttgtacaaa aaagcaggct ccccagctgg gggccgggcc gggagcctga aggacccaga
      781 tgtggctgag ctcttcttca aggatgaccc agaaaagctc ttctctgacc tccgggaaat
      841 tggccatggc agctttggag ccgtatactt tgcccgggat gtccggaata gtgaggtggt
      901 ggccatcaag aagatgtcct acagtgggaa gcagtccaat gagaaatggc aagacatcat
      961 caaggaggtg cggttcttac agaagctccg gcatcccaac accattcagt accggggctg
     1021 ttacctgagg gagcacacgg cttggctggt aatggagtat tgcctgggct cagcttctga
     1081 ccttctagaa gtgcacaaga aaccccttca ggaggtagag atcgcagctg tgacccacgg
     1141 ggcgcttcag ggcctggcat atctgcactc ccacaacatg atccataggg atgtgaaggc
     1201 tggaaacatc ctgctgtcag agccagggtt agtgaagcta ggggactttg gttctgcgtc
     1261 catcatggca cctgccaact ccttcgtggg caccccatac tggatggcac ccgaggtgat
     1321 cctggccatg gatgaggggc agtacgatgg caaagtggac gtctggtcct tggggataac
     1381 ctgcatcgag ctggctgaac ggaaaccacc gctctttaac atgaatgcga tgagtgcctt
     1441 ataccacatt gcacagaacg aatcccccgt gctccagtca ggacactggt ctgagtactt
     1501 ccggaatttt gtcgactcct gtcttcagaa aatccctcaa gacagaccaa cctcagaggt
     1561 tctcctgaag caccgctttg tgctccggga gcggccaccc acagtcatca tggacctgat
     1621 ccagaggacc aaggatgccg tgcgggagct ggacaacctg cagtaccgca agatgaagaa
     1681 gatcctgttc caagaggcac ccaacggccc tggtgccgag gccccagagg aggaagagga
     1741 ggccgagccc tacatgcacc gggccgggac tctgaccagc ctcgagagta gccactcagt
     1801 gcccagcatg tccatcagcg cctccagcca gagcagctcc gtcaacagcc tagcagatgc
     1861 ctcagacaac gaggaagagg aggaggagga ggaggaagag gaggaggagg aagaaggccc
     1921 tgaagcccgg gagatggcca tgatgcagga gggggagcac acagtcacct ctcacagctc
     1981 cattatccac cggctgccgg gctctgacaa cctatatgat gacccctacc agccagagat
     2041 aacccccagc cctctccagc cgcctgcagc cccagctccc acttccacca cctcttccgc
     2101 ccgccgccgg gcctactgcc gtaaccgaga ccactttgcc accatccgaa ccgcctccct
     2161 ggtcagccgt cagatccagg agcatgagca ggactctgcg ctgcgggagc agctgagcgg
     2221 ctataagcgg atgcgacgac agcaccagaa gcagctgctg gccctggagt cacggctgag
     2281 gggtgaacgg gaggagcaca gtgcacggct gcagcgggag cttgaggcgc agcgggctgg
     2341 ctttggggca gaggcagaaa agctggcccg gcggcaccag gccataggtg agaaggaggc
     2401 acgagctgcc caggccgagg agcggaagtt ccagcagcac atccttgggc agcagaagaa
     2461 ggagctggct gccctgctgg aggcacagaa gcggacctac aaacttcgca aggaacagct
     2521 gaaggaggag ctccaggaga accccagcac tcccaagcgg gagaaggccg agtggctgct
     2581 gcggcagaag gagcagctcc agcagtgcca ggcggaggag gaagcagggc tgctgcggcg
     2641 gcagcgccag tactttgagc tgcagtgtcg ccagtacaag cgcaagatgt tgctggctcg
     2701 gcacagcctg gaccaggacc tgctgcggga ggacctgaac aagaagcaga cccagaagga
     2761 cttggagtgt gcactgctgc ttcggcagca cgaggccacg cgggagctgg agctgcggca
     2821 gctccaggcc gtgcagcgca cgcgggctga gctcacccgc ctgcagcacc agacggagct
     2881 gggcaaccag ctggagtaca acaagcggcg tgagcaagag ttgcggcaga agcatgcggc
     2941 ccaggttcgc cagcagccca agagcctcaa agtacgtgca ggccagcgcc ccccgggcct
     3001 tccactcccc attcctgggg ctctgggccc acccaacaca ggcaccccta tagaacagca
     3061 gccctgctca cctggccagg aggcagtcct ggaccaaaga atgcttggcg aggaggagga
     3121 agcagttgga gagagaagga ttctgggaaa ggaaggggcc actttggagc ccaagcagca
     3181 gaggattctg ggggaagaat caggagcccc tagtcccagt ccacaaaaac atgggagcct
     3241 ggttgatgag gaagtttggg gtctgcctga ggagatagag gagcttaggg tgccctccct
     3301 tgtaccccag gagaggagca ttgttggcca ggaggaggct gggacatgga gcttgtgggg
     3361 gaaggaggat gagagtcttc tggatgagga gtttgagctt ggctgggtcc agggcccagc
     3421 actgactccc gtccctgagg aggaggaaga agaggaagag ggggctccga ttgggacccc
     3481 tagggatcct ggagatggtt gtccttcccc cgacatccct cctgaacccc ctccaacaca
     3541 cctgaggccc tgccctgcca gccagctccc tggactcctg tcccatggcc tcctggccgg
     3601 cctctccttt gcagtggggt cctcctctgg cctcctgccc ctcctgctgc tgctgctgct
     3661 tccattgctg gcagcccagg gtgggggtgg cctgcaggca gcgctgctgg cccttgaggt
     3721 ggggctggtg ggtctggggg cctcctacct gctcctttgt acagccctgc acctgccctc
     3781 cagtcttttc ctactcctgg cccagggtac cgcactgggg gccgtcctgg gcctgagctg
     3841 gcgccgaggc ctcatgggtg ttcccctggg ccttggagct gcctggctct tagcttggcc
     3901 aggcctagct ctacctctgg tggctatggc agcggggggc agatgggtgc ggcagcaggg
     3961 cccccgggtg cgccggggca tatctcgact ctggttgcgg gttctgctgc gcctgtcacc
     4021 catggccttc cgggccctgc agggctgtgg ggctgtgggg gaccggggtc tgtttgcact
     4081 gtaccccaaa accaacaagg atggcttccg cagccgcctg cccgtccctg ggccccggcg
     4141 gcgtaatccc cgcaccaccc aacacccatt agctctgttg gcaagggtct gggtcctgtg
     4201 caagggctgg aactggcgtc tggcacgggc cagccagggt ttagcatccc acttgccccc
     4261 gtgggccatc cacacactgg ccagctgggg cctgcttcgg ggtgaacggc ccacccgaat
     4321 cccccggcta ctaccacgca gccagcgcca gctagggccc cctgcctccc gccagccact
     4381 gccagggact ctagccgggc ggaggtcacg cacccgccag tcccgggccc tgcccccctg
     4441 gaggtaggac ccagctttct tgtacaaagt ggttgattcg gtaccagtcg actctagagg
     4501 atcccgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca
     4561 ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg
     4621 tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga
     4681 agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat
     4741 cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg
     4801 agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga
     4861 gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc
     4921 caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc
     4981 ttctgatttt aaaataacta taccagcagg aggacgtcca gacacagcat aggctacctg
     5041 gccatgccca accggtggga catttgagtt gcttgcttgg cactgtcctc tcatgcgttg
     5101 ggtccactca gtagatgcct gttgaattgg gtacgcggcc agcttggctg tggaatgtgt
     5161 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5221 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5281 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5341 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5401 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     5461 tttttggagg cctaggcttt tgcaaaaagc tcctcgagga actgaaaaac cagaaagtta
     5521 attccctata gtgagtcgta ttaaattcgt aatcatgtca tagctgtttc ctgtgtgaaa
     5581 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5641 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5701 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5761 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5821 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5881 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5941 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6001 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6061 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6121 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6181 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6241 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6301 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6361 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6421 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6481 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6541 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6601 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6661 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6721 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6781 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6841 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6901 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6961 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7021 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7081 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7141 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7201 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7261 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7321 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7381 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7441 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7501 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7561 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7621 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7681 gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc
     7741 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     7801 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     7861 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     7921 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     7981 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     8041 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     8101 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     8161 aatttccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     8221 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     8281 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct gatctataca
     8341 ttgaatcaat attggcaatt agccatatta gtcattggtt atatagcata aatcaatatt
     8401 ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta tattggctca
     8461 tgtccaatat gaccgccatg tt
//
