LOCUS       sequence-340495-        5943 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression of FLAG-tagged STK3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5943)
  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 5943)
  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-STK3
FEATURES             Location/Qualifiers
     source          1..5943
                     /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"
     RBS             1710..1718
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     protein_bind    complement(2534..2558)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2591..3213
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2790..2809)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3237..3257)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3242..3571
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3422..3557
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3609..3628)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3610..3628)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3642..3658)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3642..3658)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3655..3677)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3666..3682
                     /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(3690..3696,3697..3714,3715..3720))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3735..3756
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3873..3890)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4044..4632)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4124..4143)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4803..5594,5595..5663))
                     /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     5426..5445
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5664..5768)
                     /gene="bla"
                     /label=AmpR promoter
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 agcagccgcc ggcgcctaag
     1081 agtaaactaa aaaagctgag tgaagacagt ttgactaagc agcctgaaga agtttttgat
     1141 gtattagaga agcttggaga agggtcttat ggaagtgtat ttaaagcaat acacaaggaa
     1201 tccggtcaag ttgtcgcaat taaacaagta cctgttgaat cagatcttca ggaaataatc
     1261 aaagaaattt ccataatgca gcaatgtgac agcccatatg ttgtaaagta ctatggcagt
     1321 tattttaaga atacagacct ctggattgtt atggagtact gtggcgctgg ctctgtctca
     1381 gacataatta gattacgaaa caagacatta atagaagatg aaattgcaac cattcttaaa
     1441 tctacattga aaggactaga atatttgcac tttatgagaa aaatacacag agatataaaa
     1501 gctggaaata ttctcctcaa tacagaagga catgcaaaat tggcagattt tggagtggct
     1561 ggtcagttaa cagatacaat ggcaaaacgc aatactgtaa taggaactcc attttggatg
     1621 gctcctgagg tgattcaaga aataggctat aactgtgtgg ccgacatctg gtcccttggc
     1681 attacttcta tagaaatggc tgaaggaaaa cctccttatg ctgatataca tccaatgagg
     1741 gctattttta tgattcccac aaatccacca ccaacattca gaaagccaga actttggtcc
     1801 gatgatttca ccgattttgt taaaaagtgt ttggtgaaga atcctgagca gagagctact
     1861 gcaacacaac ttttacagca tccttttatc aagaatgcca aacctgtatc aatattaaga
     1921 gacctgatca cagaagctat ggagatcaaa gctaaaagac atgaggaaca gcaacgagaa
     1981 ttggaagagg aagaagaaaa ttcggatgaa gatgagctgg attcccacac catggtgaag
     2041 actagtgtgg agagtgtggg caccatgcgg gccacaagca cgatgagtga aggggcccag
     2101 accatgattg aacataatag cacgatgttg gaatccgact tggggaccat ggtgataaac
     2161 agtgaggatg aggaagaaga agatggaact atgaaaagaa atgcaacctc accacaagta
     2221 caaagaccat ctttcatgga ctactttgat aagcaagact tcaagaataa gagtcacgaa
     2281 aactgtaatc agaacatgca tgagcccttc cctatgtcca aaaacgtttt tcctgataac
     2341 tggaaagttc ctcaagatgg agactttgac tttttgaaaa atctaagttt agaagaacta
     2401 cagatgcggt taaaagcact ggaccccatg atggaacggg agatagaaga acttcgtcag
     2461 agatacactg cgaaaagaca gcccattctg gatgcgatgg atgcaaagaa aagaaggcag
     2521 caaaactttt gagacccagc tttcttgtac aaagtggttg attcggtacc agtcgactct
     2581 agaggatccc gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt
     2641 tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga
     2701 ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt
     2761 tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc
     2821 acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc
     2881 tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg
     2941 gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat
     3001 ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc
     3061 tgtccttctg attttaaaat aactatacca gcaggaggac gtccagacac agcataggct
     3121 acctggccat gcccaaccgg tgggacattt gagttgcttg cttggcactg tcctctcatg
     3181 cgttgggtcc actcagtaga tgcctgttga attgggtacg cggccagctt ggctgtggaa
     3241 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3301 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3361 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3421 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3481 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3541 aggctttttt ggaggcctag gcttttgcaa aaagctcctc gaggaactga aaaaccagaa
     3601 agttaattcc ctatagtgag tcgtattaaa ttcgtaatca tgtcatagct gtttcctgtg
     3661 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     3721 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     3781 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3841 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3901 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3961 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4021 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4081 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4141 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4201 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4261 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4321 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4381 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4441 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4501 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4561 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4621 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4681 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4741 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4801 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4861 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4921 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4981 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5041 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5101 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5161 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5221 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5281 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5341 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5401 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5461 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5521 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5581 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5641 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5701 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5761 gttccgcgca catttccccg aaaagtgsnc wcytgwyscr mmctgkarcr acactcaacc
     5821 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     5881 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     5941 ttt
//
