LOCUS       sequence-340591-        8041 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression of FLAG-tagged LATS2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8041)
  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 8041)
  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-LATS2
FEATURES             Location/Qualifiers
     source          1..8041
                     /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(4325..4349)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    4382..5004
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4581..4600)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(5028..5048)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5033..5362
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5213..5348
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5400..5419)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5401..5419)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5433..5449)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5433..5449)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5446..5468)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5457..5473
                     /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(5481..5487,5488..5505,5506..5511))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5526..5547
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5664..5681)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5835..6423)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5915..5934)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6594..7385,7386..7454))
                     /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     7217..7236
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7455..7559)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7586..8014
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7673..7692)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7883..7904
                     /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 gcaggctcta ggccaaagac ttttcctgcc
     1081 acgacttatt ctggaaatag ccggcagcga ctgcaagaga ttcgtgaggg gttaaagcag
     1141 ccatccaagt cttcggttca ggggctaccc gcaggaccaa acagtgacac ttccctggat
     1201 gccaaagtcc tggggagcaa agatgccacc aggcagcagc agcagatgag agccacccca
     1261 aagttcggac cttatcagaa agccttgagg gaaatcagat attccttgtt gccttttgct
     1321 aatgaatcgg gcacctctgc agctgcagaa gtgaaccggc aaatgctgca ggaactggtg
     1381 aacgcaggat gcgaccagga gatggctggc cgagctctca agcagactgg cagcaggagc
     1441 atcgaggccg ccctggagta catcagcaag atgggctacc tggacccgag gaatgagcag
     1501 attgtgcggg tcattaagca gacctcccca ggaaaggggc tcatgccaac cccagtgacg
     1561 cggaggccca gcttcgaagg aaccggcgat tcgtttgcgt cctaccacca gctgagcggt
     1621 accccctacg agggcccaag cttcggcgct gacggcccca cggcgctgga ggagatgccg
     1681 cggccgtacg tggactacct tttccccgga gtcggccccc acgggcccgg ccaccagcac
     1741 cagcacccac ccaagggcta cggtgccagc gtagaggcag caggggcaca cttcccgctg
     1801 cagggcgcgc actacgggcg gccgcacctg ctggtgcctg gggaacccct gggctacgga
     1861 gtgcagcgca gcccctcctt ccagagcaag acgccgccgg agaccggggg ttacgccagc
     1921 ctgcccacga agggccaggg aggaccgcca ggcgccggcc tcgctttccc accccctgcc
     1981 gccgggctct acgtgccgca cccacaccac aagcaggccg gtcccgtggc ccaccagctg
     2041 catgtgctgg gctcccgcag ccaggtgttc gccagcgaca gccccccgca gagcctgctc
     2101 actccctcgc ggaacagcct caacgtggac ctgtatgaat tgagcagcac ctccgtccag
     2161 cagtggccgg ctgccaccct ggcccgccgg gactccctgc agaagccggg cctggaggcg
     2221 ccgccgcgcg cgcacgtggc cttccggcct gactgcccag tgcccagcag gaccaactcc
     2281 ttcaacagcc accagccgcg gcccggtccg cctggcaagg ccgagccctc cctgcccgcc
     2341 cccaacaccg tgacggctgt cacggccgcg cacatcttgc acccggtgaa gagcgtgcgt
     2401 gtgctgaggc cggagccgca gacggctgtg gggccctcgc accccgcctg ggtgcccgcg
     2461 cctgccccgg cccccgcccc cgcccccgcc ccggctgcgg agggcttgga cgccaaggag
     2521 gagcatgccc tggcgctggg cggcgcaggc gccttcccgc tggacgtgga gtacggaggc
     2581 ccagaccgga ggtgcccgcc tccgccctac ccgaagcacc tgctgctgcg cagcaagtcg
     2641 gagcagtacg acctggacag cctgtgcgca ggcatggagc agagcctccg tgcgggcccc
     2701 aacgagcccg agggcggcga caagagccgc aaaagcgcca agggggacaa aggcggaaag
     2761 gataaaaagc agattcagac ctctcccgtt cccgtccgca aaaacagcag agacgaagag
     2821 aagagagagt cacgcatcaa gagctactcg ccatacgcct ttaagttctt catggagcag
     2881 cacgtggaga atgtcatcaa aacctaccag cagaaggtta accggaggct gcagctggag
     2941 caagaaatgg ccaaagctgg actctgtgaa gctgagcagg agcagatgcg gaagatcctc
     3001 taccagaaag agtctaatta caacaggtta aagagggcca agatggacaa gtctatgttt
     3061 gtcaagatca aaaccctggg gatcggtgcc tttggagaag tgtgccttgc ttgtaaggtg
     3121 gacactcacg ccctgtacgc catgaagacc ctaaggaaaa aggatgtcct gaaccggaat
     3181 caggtggccc acgtcaaggc cgagagggac atcctggccg aggcagacaa tgagtgggtg
     3241 gtcaaactct actactcctt ccaagacaaa gacagcctgt actttgtgat ggactacatc
     3301 cctggtgggg acatgatgag cctgctgatc cggatggagg tcttccctga gcacctggcc
     3361 cggttctaca tcgcagagct gactttggcc attgagagtg tccacaagat gggcttcatc
     3421 caccgagaca tcaagcctga taacattttg atagatctgg atggtcacat taaactcaca
     3481 gatttcggcc tctgcactgg gttcaggtgg actcacaatt ccaaatatta ccagaaaggg
     3541 agccatgtca gacaggacag catggagccc agcgacctct gggatgatgt gtctaactgt
     3601 cggtgtgggg acaggctgaa gaccctagag cagagggcgc ggaagcagca ccagaggtgc
     3661 ctggcacatt cactggtggg gactccaaac tacatcgcac ccgaggtgct cctccgcaaa
     3721 gggtacactc aactctgtga ctggtggagt gttggagtga ttctcttcga gatgctggtg
     3781 gggcagccgc cctttttggc acctactccc acagaaaccc agctgaaggt gatcaactgg
     3841 gagaacacgc tccacattcc agcccaggtg aagctgagcc ctgaggccag ggacctcatc
     3901 accaagctgt gctgctccgc agaccaccgc ctggggcgga atggggccga tgacctgaag
     3961 gcccacccct tcttcagcgc cattgacttc tccagtgaca tccggaagca gccagccccc
     4021 tacgttccca ccatcagcca ccccatggac acctcgaatt tcgaccccgt agatgaagaa
     4081 agcccttgga acgatgccag cgaaggtagc accaaggcct gggacacact cacctcgccc
     4141 aataacaagc atcctgagca cgcattttac gaattcacct tccgaaggtt ctttgatgac
     4201 aatggctacc cctttcgatg cccaaagcct tcaggagcag aagcttcaca ggctgagagc
     4261 tcagatttag aaagctctga tctggtggat cagactgaag gctgccagcc tgtgtacgtg
     4321 taggacccag ctttcttgta caaagtggtt gattcggtac cagtcgactc tagaggatcc
     4381 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc
     4441 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc
     4501 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac
     4561 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg
     4621 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt
     4681 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg
     4741 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc
     4801 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct
     4861 gattttaaaa taactatacc agcaggagga cgtccagaca cagcataggc tacctggcca
     4921 tgcccaaccg gtgggacatt tgagttgctt gcttggcact gtcctctcat gcgttgggtc
     4981 cactcagtag atgcctgttg aattgggtac gcggccagct tggctgtgga atgtgtgtca
     5041 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5101 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     5161 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     5221 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     5281 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     5341 tggaggccta ggcttttgca aaaagctcct cgaggaactg aaaaaccaga aagttaattc
     5401 cctatagtga gtcgtattaa attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt
     5461 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5521 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5581 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5641 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5701 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5761 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5821 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5881 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5941 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6001 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6061 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6121 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6181 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6241 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     6301 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6361 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6421 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6481 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6541 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6601 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6661 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6721 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6781 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6841 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6901 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6961 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7021 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7081 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7141 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7201 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7261 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7321 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7381 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7441 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7501 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7561 acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt
     7621 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     7681 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     7741 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     7801 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     7861 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     7921 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     7981 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     8041 t
//
