LOCUS       sequence-340436-        6238 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression of FLAG-tagged STK4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6238)
  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 6238)
  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-STK4
FEATURES             Location/Qualifiers
     source          1..6238
                     /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(2522..2546)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2579..3201
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2778..2797)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3225..3245)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3230..3559
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3410..3545
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3597..3616)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3598..3616)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3630..3646)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3630..3646)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3643..3665)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3654..3670
                     /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(3678..3684,3685..3702,3703..3708))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3723..3744
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3861..3878)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4032..4620)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4112..4131)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4791..5582,5583..5651))
                     /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     5414..5433
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5652..5756)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5783..6211
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5870..5889)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6080..6101
                     /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 gcaggctctg agacggtaca gctgaggaac
     1081 ccgccgcgcc ggcagctgaa aaagttggat gaagatagtt taaccaaaca accagaagaa
     1141 gtatttgatg tcttagagaa acttggagaa gggtcctatg gcagcgtata caaagctatt
     1201 cataaagaga ccggccagat tgttgctatt aagcaagttc ctgtggaatc agacctccag
     1261 gagataatca aagaaatctc tataatgcag caatgtgaca gccctcatgt agtcaaatat
     1321 tatggcagtt attttaagaa cacagactta tggatcgtta tggagtactg tggggctggt
     1381 tctgtatctg atatcattcg attacgaaat aaaacgttaa cagaagatga aatagctaca
     1441 atattacaat caactcttaa gggacttgaa taccttcatt ttatgagaaa aatacaccga
     1501 gatatcaagg caggaaatat tttgctaaat acagaaggac atgcaaaact tgcagatttt
     1561 ggggtagcag gtcaacttac agataccatg gccaagcgga atacagtgat aggaacacca
     1621 ttttggatgg ctccagaagt gattcaggaa attggataca actgtgtagc agacatctgg
     1681 tccctgggaa taactgccat agaaatggct gaaggaaagc ccccttatgc tgatatccat
     1741 ccaatgaggg caatcttcat gattcctaca aatcctcctc ccacattccg aaaaccagag
     1801 ctatggtcag ataactttac agattttgtg aaacagtgtc ttgtaaagag ccctgagcag
     1861 agggccacag ccactcagct cctgcagcac ccatttgtca ggagtgccaa aggagtgtca
     1921 atactgcgag acttaattaa tgaagccatg gatgtgaaac tgaaacgcca ggaatcccag
     1981 cagcgggaag tggaccagga cgatgaagaa aactcagaag aggatgaaat ggattctggc
     2041 acgatggttc gagcagtggg tgatgagatg ggcactgtcc gagtagccag caccatgact
     2101 gatggagcca atactatgat tgagcacgat gacacgttgc catcacaact gggcaccatg
     2161 gtgatcaatg cagaggatga ggaagaggaa ggaactatga aaagaaggga tgagaccatg
     2221 cagcctgcga aaccatcctt tcttgaatat tttgaacaaa aagaaaagga aaaccagatc
     2281 aacagctttg gcaagagtgt acctggtcca ctgaaaaatt cttcagattg gaaaatacca
     2341 caggatggag actacgagtt tcttaagagt tggacagtgg aggaccttca gaagaggctc
     2401 ttggccctgg accccatgat ggagcaggag attgaagaga tccggcagaa gtaccagtcc
     2461 aagcggcagc ccatcctgga tgccatagag gctaagaaga gacggcaaca aaacttctga
     2521 gacccagctt tcttgtacaa agtggttgat tcggtaccag tcgactctag aggatcccgg
     2581 gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt
     2641 gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt
     2701 ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac
     2761 ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct
     2821 cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt
     2881 gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg
     2941 tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg
     3001 gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat
     3061 tttaaaataa ctataccagc aggaggacgt ccagacacag cataggctac ctggccatgc
     3121 ccaaccggtg ggacatttga gttgcttgct tggcactgtc ctctcatgcg ttgggtccac
     3181 tcagtagatg cctgttgaat tgggtacgcg gccagcttgg ctgtggaatg tgtgtcagtt
     3241 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3301 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3361 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3421 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3481 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3541 aggcctaggc ttttgcaaaa agctcctcga ggaactgaaa aaccagaaag ttaattccct
     3601 atagtgagtc gtattaaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat
     3661 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     3721 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     3781 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3841 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3901 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3961 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4021 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4081 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4141 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4201 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4261 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4321 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4381 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4441 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4501 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4561 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4621 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4681 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4741 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4801 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4861 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4921 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4981 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5041 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5101 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5161 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5221 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5281 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5341 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5401 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5461 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5521 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5581 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5641 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5701 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5761 tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     5821 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     5881 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     5941 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     6001 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     6061 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     6121 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     6181 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaattt
//
