LOCUS       sequence-340454-        5425 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression of FLAG-tagged MOB1A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5425)
  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 5425)
  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-MOB1A
FEATURES             Location/Qualifiers
     source          1..5425
                     /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(1709..1733)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1766..2388
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1965..1984)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2412..2432)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2417..2746
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2597..2732
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2784..2803)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2785..2803)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2817..2833)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2817..2833)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2830..2852)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2841..2857
                     /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(2865..2871,2872..2889,2890..2895))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2910..2931
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3048..3065)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3219..3807)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3299..3318)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3978..4769,4770..4838))
                     /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     4601..4620
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4839..4943)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4970..5398
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5057..5076)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5267..5288
                     /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 gcaggctcca gcttcctctt cagcagccgc
     1081 tcttctaaaa cattcaaacc aaagaagaat atccctgaag gatctcatca gtatgaactc
     1141 ttaaaacatg cagaagcaac tctaggaagt gggaatctga gacaagctgt tatgttgcct
     1201 gagggagagg atctcaatga atggattgct gtgaacactg tggatttctt taaccagatc
     1261 aacatgttat atggaactat tacagaattc tgcactgaag caagctgtcc agtcatgtct
     1321 gcaggtccga gatatgaata tcactgggca gatggtacta atattaaaaa gccaatcaaa
     1381 tgttctgcac caaaatacat tgactatttg atgacttggg ttcaagatca gcttgatgat
     1441 gaaactcttt ttccttctaa gattggtgtc ccatttccca aaaactttat gtctgtggca
     1501 aagactattc taaagcgtct gttcagggtt tatgcccata tttatcacca gcactttgat
     1561 tctgtgatgc agctgcaaga ggaggcccac ctcaacacct cctttaagca ctttattttc
     1621 tttgttcagg agtttaatct gattgatagg cgtgagctgg cacctcttca agaattaata
     1681 gagaaacttg gatcaaaaga cagataagac ccagctttct tgtacaaagt ggttgattcg
     1741 gtaccagtcg actctagagg atcccgggtg gcatccctgt gacccctccc cagtgcctct
     1801 cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg
     1861 catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg
     1921 gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag
     1981 ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat
     2041 tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta
     2101 atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc
     2161 ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac
     2221 cactgctccc ttccctgtcc ttctgatttt aaaataacta taccagcagg aggacgtcca
     2281 gacacagcat aggctacctg gccatgccca accggtggga catttgagtt gcttgcttgg
     2341 cactgtcctc tcatgcgttg ggtccactca gtagatgcct gttgaattgg gtacgcggcc
     2401 agcttggctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     2461 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     2521 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     2581 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     2641 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     2701 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcctcgagga
     2761 actgaaaaac cagaaagtta attccctata gtgagtcgta ttaaattcgt aatcatgtca
     2821 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     2881 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     2941 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     3001 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     3061 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3121 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3181 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3241 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3301 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3361 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3421 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3481 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3541 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3601 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3661 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3721 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3781 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3841 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3901 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3961 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4021 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4081 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4141 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4201 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4261 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4321 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4381 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4441 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4501 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4561 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4621 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4681 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4741 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4801 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4861 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4921 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt
     4981 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     5041 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     5101 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     5161 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     5221 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     5281 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     5341 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     5401 aacaaaatat taacgcttac aattt
//
