LOCUS       sequence-402477-        8959 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Myc-tagged LATS1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8959)
  AUTHORS   Bao Y, Nakagawa K, Yang Z, Ikeda M, Withanage K, Ishigami-Yuasa M, 
            Okuno Y, Hata S, Nishina H, Hata Y
  TITLE     A cell-based assay to screen stimulators of the Hippo pathway 
            reveals the inhibitory effect of dobutamine on the YAP-dependent 
            gene transcription.
  JOURNAL   J Biochem. 2011 Aug;150(2):199-208. doi: 10.1093/jb/mvr063. Epub 
            2011 May 17.
  PUBMED    21586534
REFERENCE   2  (bases 1 to 8959)
  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"; doi: "10.1093/jb/mvr063";
            journalName: "J Biochem"; date: "2011-08"; volume: "150"; issue: 
            "2"; pages: "199-208"
COMMENT     Sequence Label: pClneoMyc-LATS1
FEATURES             Location/Qualifiers
     source          1..8959
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1103..1132
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    4663..4784
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4674..4693)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4728..4747
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4970..5425
                     /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"
     promoter        5542..5899
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5542..5733
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5565..5585)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5750..5885
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5950..6744
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6004..6023)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6614..6633
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6808..6856
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6897..6916)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7016..7038
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7076..7094)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7162..7266
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7267..7335,7336..8127)
                     /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     complement(7485..7504)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8298..8886
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8787..8806
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctaga ccccccaaca tggagcagaa gcttatcagc gaggaggacc tgctcgagga
     1141 attcacgcgt atgaagagga gtgaaaagcc agaaggatat agacaaatga ggcctaagac
     1201 ctttcctgcc agtaactata ctgtcagtag ccggcaaatg ttacaagaaa ttcgggaatc
     1261 ccttaggaat ttatctaaac tatctgatgc tgctaaggct gagcataaca tgagtaaaat
     1321 gtcaaccgaa gatcctcgac aagtcagaaa tccacccaaa tttgggacgc atcataaagc
     1381 cttgcaggaa attcgaaact ctctgcttcc atttgcaaat gaaacaaatt cttctcggag
     1441 tacttcagaa gttaatccac aaatgcttca agacttgcaa gctgctggat ttgatgagga
     1501 tatggttata caagctcttc agaaaactaa caacagaagt atagaagcag caattgaatt
     1561 cattagtaaa atgagttacc aagatcctcg acgagagcag atggctgcag cagctgccag
     1621 acctattaat gccagcatga aaccagggaa tgtgcagcaa tcagttaacc gcaaacagag
     1681 ctggaaaggt tctaaagaat ccttagttcc tcagaggcat ggcccgccac taggagaaag
     1741 tgtggcctat cattctgaga gtcccaactc acagacagat gtaggaagac ctttgtctgg
     1801 atctggtata tcagcatttg ttcaagctca ccctagcaac ggacagagag tgaacccccc
     1861 accaccacct caagtaagga gtgttactcc tccaccacct ccaagaggcc agactccccc
     1921 tccaagaggt acaactccac ctcccccttc atgggaacca aactctcaaa caaagcgcta
     1981 ttctggaaac atggaatacg taatctcccg aatctctcct gtcccacctg gggcatggca
     2041 agagggctat cctccaccac ctctcaacac ttcccccatg aatcctccta atcaaggaca
     2101 gagaggcatt agttctgttc ctgttggcag acaaccaatc atcatgcaga gttctagcaa
     2161 atttaacttt ccatcaggga gacctggaat gcagaatggt actggacaaa ctgatttcat
     2221 gatacaccaa aatgttgtcc ctgctggcac tgtgaatcgg cagccaccac ctccatatcc
     2281 tctgacagca gctaatggac aaagcccttc tgctttacaa acagggggat ctgctgctcc
     2341 ttcgtcatat acaaatggaa gtattcctca gtctatgatg gtgccaaaca gaaatagtca
     2401 taacatggaa ctatataaca ttagtgtacc tggactgcaa acaaattggc ctcagtcatc
     2461 ttctgctcca gcccagtcat ccccgagcag tgggcatgaa atccctacat ggcaacctaa
     2521 cataccagtg aggtcaaatt cttttaataa cccattagga aatagagcaa gtcactctgc
     2581 taattctcag ccttctgcta caacagtcac tgcaattaca ccagctccta ttcaacagcc
     2641 tgtgaaaagt atgcgtgtat taaaaccaga gctacagact gctttagcac ctacacaccc
     2701 ttcttggata ccacagccaa ttcaaactgt tcaacccagt ccttttcctg agggaaccgc
     2761 ttcaaatgtg actgtgatgc cacctgttgc tgaagctcca aactatcaag gaccaccacc
     2821 accctaccca aaacatctgc tgcaccaaaa cccatctgtt cctccatacg agtcaatcag
     2881 taagcctagc aaagaggatc agccaagctt gcccaaggaa gatgagagtg aaaagagtta
     2941 tgaaaatgtt gatagtgggg ataaagaaaa gaaacagatt acaacttcac ctattactgt
     3001 taggaaaaac aagaaagatg aagagcgaag ggaatctcgt attcaaagtt attctcctca
     3061 agcatttaaa ttctttatgg agcaacatgt agaaaatgta ctcaaatctc atcagcagcg
     3121 tctacatcgt aaaaaacaat tagagaatga aatgatgcgg gttggattat ctcaagatgc
     3181 ccaggatcaa atgagaaaga tgctttgcca aaaagaatct aattacatcc gtcttaaaag
     3241 ggctaaaatg gacaagtcta tgtttgtgaa gataaagaca ctaggaatag gagcatttgg
     3301 tgaagtctgt ctagcaagaa aagtagatac taaggctttg tatgcaacaa aaactcttcg
     3361 aaagaaagat gttcttcttc gaaatcaagt cgctcatgtt aaggctgaga gagatatcct
     3421 ggctgaagct gacaatgaat gggtagttcg tctatattat tcattccaag ataaggacaa
     3481 tttatacttt gtaatggact acattcctgg gggtgatatg atgagcctat taattagaat
     3541 gggcatcttt ccagaaagtc tggcacgatt ctacatagca gaacttacct gtgcagttga
     3601 aagtgttcat aaaatgggtt ttattcatag agatattaaa cctgataata ttttgattga
     3661 tcgtgatggt catattaaat tgactgactt tggcctctgc actggcttca gatggacaca
     3721 cgattctaag tactatcaga gtggtgacca tccacggcaa gatagcatgg atttcagtaa
     3781 tgaatggggg gatccctcaa gctgtcgatg tggagacaga ctgaagccat tagagcggag
     3841 agctgcacgc cagcaccagc gatgtctagc acattctttg gttgggactc ccaattatat
     3901 tgcacctgaa gtgttgctac gaacaggata cacacagttg tgtgattggt ggagtgttgg
     3961 tgttattctt tttgaaatgt tggtgggaca acctcctttc ttggcacaaa caccattaga
     4021 aacacaaatg aaggttatca actggcaaac atctcttcac attccaccac aagctaaact
     4081 cagtcctgaa gcttctgatc ttattattaa actttgccga ggacccgaag atcgcttagg
     4141 caagaatggt gctgatgaaa taaaagctca tccatttttt aaaacaattg acttctccag
     4201 tgacctgaga cagcagtctg cttcatacat tcctaaaatc acacacccaa cagatacatc
     4261 aaattttgat cctgttgatc ctgataaatt atggagtgat gataacgagg aagaaaatgt
     4321 aaatgacact ctcaatggat ggtataaaaa tggaaagcat cctgaacatg cattctatga
     4381 atttaccttc cgaaggtttt ttgatgacaa tggctaccca tataattatc cgaagcctat
     4441 tgaatatgaa tacattaatt cacaaggctc agagcagcag tcggatgaag atgatcaaaa
     4501 cacaggctca gagattaaaa atcgcgatct agtatatgtt taacacacta gtaaataaat
     4561 gtaatgagga tttgtaaaag ggcctgaaat gcgaggtgtt ttgaggtacc gtcgacgcgg
     4621 ccgcttccct ttagtgaggg ttaatgcttc gagcagacat gataagatac attgatgagt
     4681 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     4741 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     4801 ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc
     4861 tctacaaatg tggtaaaatc cgataaggat cgatccgggc tggcgtaata gcgaagaggc
     4921 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga 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 aatttcctga tgcggtattt tctccttacg catctgtgcg
     5461 gtatttcaca ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag
     5521 aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     5581 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     5641 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5701 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     5761 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     5821 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     5881 aggcctaggc ttttgcaaaa agcttgattc ttctgacaca acagtctcga acttaaggct
     5941 agagccacca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     6001 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     6061 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     6121 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     6181 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     6241 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     6301 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     6361 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     6421 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     6481 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     6541 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     6601 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     6661 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     6721 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     6781 cgcccaacct gccatcacga tggccgcaat aaaatatctt tattttcatt acatctgtgt
     6841 gttggttttt tgtgtgaatc gatagcgata aggatccgcg tatggtgcac tctcagtaca
     6901 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     6961 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     7021 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     7081 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     7141 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     7201 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     7261 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     7321 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     7381 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     7441 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     7501 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     7561 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     7621 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     7681 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     7741 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     7801 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     7861 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     7921 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     7981 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     8041 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     8101 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     8161 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     8221 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     8281 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     8341 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     8401 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     8461 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     8521 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     8581 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     8641 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     8701 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     8761 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     8821 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     8881 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     8941 cacatggctc gacagatct
//
