LOCUS       sequence-340300-        8167 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression of FLAG-tagged LATS1 .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8167)
  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 8167)
  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-LATS1
FEATURES             Location/Qualifiers
     source          1..8167
                     /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(4451..4475)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    4508..5130
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4707..4726)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(5154..5174)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5159..5488
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5339..5474
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5526..5545)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5527..5545)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5559..5575)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5559..5575)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5572..5594)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5583..5599
                     /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(5607..5613,5614..5631,5632..5637))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5652..5673
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5790..5807)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5961..6549)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6041..6060)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6720..7511,7512..7580))
                     /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     7343..7362
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7581..7685)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7712..8140
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7799..7818)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8009..8030
                     /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 agaggagtga aaagccagaa
     1081 ggatatagac aaatgaggcc taagaccttt cctgccagta actatactgt cagtagccgg
     1141 caaatgttac aagaaattcg ggaatccctt aggaatttat ctaaaccatc tgatgctgct
     1201 aaggctgagc ataacatgag taaaatgtca accgaagatc ctcgacaagt cagaaatcca
     1261 cccaaatttg ggacgcatca taaagccttg caggaaattc gaaactctct gcttccattt
     1321 gcaaatgaaa caaattcttc tcggagtact tcagaagtta atccacaaat gcttcaagac
     1381 ttgcaagctg ctggatttga tgaggatatg gttatacaag ctcttcagaa aactaacaac
     1441 agaagtatag aagcagcaat tgaattcatt agtaaaatga gttaccaaga tcctcgacga
     1501 gagcagatgg ctgcagcagc tgccagacct attaatgcca gcatgaaacc agggaatgtg
     1561 cagcaatcag ttaaccgcaa acagagctgg aaaggttcta aagaatcctt agttcctcag
     1621 aggcatggcc cgccactagg agaaagtgtg gcctatcatt ctgagagtcc caactcacag
     1681 acagatgtag gaagaccttt gtctggatct ggtatatcag catttgttca agctcaccct
     1741 agcaacggac agagagtgaa ccccccacca ccacctcaag taaggagtgt tactcctcca
     1801 ccacctccaa gaggccagac tccccctcca agaggtacaa ctccacctcc cccttcatgg
     1861 gaaccaaact ctcaaacaaa gcgctattct ggaaacatgg aatacgtaat ctcccgaatc
     1921 tctcctgtcc cacctggggc atggcaagag ggctatcctc caccacctct caacacttcc
     1981 cccatgaatc ctcctaatca aggacagaga ggcattagtt ctgttcctgt tggcagacaa
     2041 ccaatcatca tgcagagttc tagcaaattt aactttccat cagggagacc tggaatgcag
     2101 aatggtactg gacaaactga tttcatgata caccaaaatg ttgtccctgc tggcactgtg
     2161 aatcggcagc caccacctcc atatcctctg acagcagcta atggacaaag cccttctgct
     2221 ttacaaacag ggggatctgc tgctccttcg tcatatacaa atggaagtat tcctcagtct
     2281 atgatggtgc caaacagaaa tagtcataac atggaactat ataacattag tgtacctgga
     2341 ctgcaaacaa attggcctca gtcatcttct gctccagccc agtcatcccc gagcagtggg
     2401 catgaaatcc ctacatggca acctaacata ccagtgaggt caaattcttt taataaccca
     2461 ttaggaaata gagcaagtca ctctgctaat tctcagcctt ctgctacaac agtcactgca
     2521 attacaccag ctcctattca acagcctgtg aaaagtatgc gtgtattaaa accagagcta
     2581 cagactgctt tagcacctac acacccttct tggataccac agccaattca aactgttcaa
     2641 cccagtcctt ttcctgaggg aaccgcttca aatgtgactg tgatgccacc tgttgctgaa
     2701 gctccaaact atcaaggacc accaccaccc tacccaaaac atctgctgca ccaaaaccca
     2761 tctgttcctc catacgagtc aatcagtaag cctagcaaag aggatcagcc aagcttgccc
     2821 aaggaagatg agagtgaaaa gagttatgaa aatgttgata gtggggataa agaaaagaaa
     2881 cagattacaa cttcacctat tactgttagg aaaaacaaga aagatgaaga gcgaagggaa
     2941 tctcgtattc aaagttattc tcctcaagca tttaaattct ttatggagca acatgtagaa
     3001 aatgtactca aatctcatca gcagcgtcta catcgtaaaa aacaattaga gaatgaaatg
     3061 atgcgggttg gattatctca agatgcccag gatcaaatga gaaagatgct ttgccaaaaa
     3121 gaatctaatt acatccgtct taaaagggct aaaatggaca agtctatgtt tgtgaagata
     3181 aagacactag gaataggagc atttggtgaa gtctgtctag caagaaaagt agatactaag
     3241 gctttgtatg caacaaaaac tcttcgaaag aaagatgttc ttcttcgaaa tcaagtcgct
     3301 catgttaagg ctgagagaga tatcctggct gaagctgaca atgaatgggt agttcgtcta
     3361 tattattcat tccaagataa ggacaattta tactttgtaa tggactacat tcctgggggt
     3421 gatatgatga gcctattaat tagaatgggc atctttccag aaagtctggc acgattctac
     3481 atagcagaac ttacctgtgc agttgaaagt gttcataaaa tgggttttat tcatagagat
     3541 attaaacctg ataatatttt gattgatcgt gatggtcata ttaaattgac tgactttggc
     3601 ctctgcactg gcttcagatg gacacacgat tctaagtact atcagagtgg tgaccatcca
     3661 cggcaagata gcatggattt cagtaatgaa tggggggatc cctcaagctg tcgatgtgga
     3721 gacagactga agccattaga gcggagagct gcacgccagc accagcgatg tctagcacat
     3781 tctttggttg ggactcccaa ttatattgca cctgaagtgt tgctacgaac aggatacaca
     3841 cagttgtgtg attggtggag tgttggtgtt attctttttg aaatgttggt gggacaacct
     3901 cctttcttgg cacaaacacc attagaaaca caaatgaagg ttatcaactg gcaaacatct
     3961 cttcacattc caccacaagc taaactcagt cctgaagctt ctgatcttat tattaaactt
     4021 tgccgaggac ccgaagatcg cttaggcaag aatggtgctg atgaaataaa agctcatcca
     4081 ttttttaaaa caattgactt ctccagtgac ctgagacagc agtctgcttc atacattcct
     4141 aaaatcacac acccaacaga tacatcaaat tttgatcctg ttgatcctga taaattatgg
     4201 agtgatgata acgaggaaga aaatgtaaat gacactctca atggatggta taaaaatgga
     4261 aagcatcctg aacatgcatt ctatgaattt accttccgaa ggttttttga tgacaatggc
     4321 tacccatata attatccgaa gcctattgaa tatgaataca ttaattcaca aggctcagag
     4381 cagcagtcgg atgaagatga tcaaaacaca ggctcagaga ttaaaaatcg cgatctagta
     4441 tatgtttaag acccagcttt cttgtacaaa gtggttgatt cggtaccagt cgactctaga
     4501 ggatcccggg tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc
     4561 cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta
     4621 ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg
     4681 gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca
     4741 atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc
     4801 cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta
     4861 gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta
     4921 cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt
     4981 ccttctgatt ttaaaataac tataccagca ggaggacgtc cagacacagc ataggctacc
     5041 tggccatgcc caaccggtgg gacatttgag ttgcttgctt ggcactgtcc tctcatgcgt
     5101 tgggtccact cagtagatgc ctgttgaatt gggtacgcgg ccagcttggc tgtggaatgt
     5161 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     5221 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     5281 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     5341 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     5401 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     5461 cttttttgga ggcctaggct tttgcaaaaa gctcctcgag gaactgaaaa accagaaagt
     5521 taattcccta tagtgagtcg tattaaattc gtaatcatgt catagctgtt tcctgtgtga
     5581 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     5641 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     5701 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5761 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5821 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5881 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5941 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6001 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6061 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6121 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6181 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6241 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6301 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6361 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     6421 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6481 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6541 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6601 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6661 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6721 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6781 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6841 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6901 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6961 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7021 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7081 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7141 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7201 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7261 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7321 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7381 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7441 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7501 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7561 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7621 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7681 ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg
     7741 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     7801 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     7861 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     7921 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     7981 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     8041 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     8101 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     8161 acaattt
//
