LOCUS       sequence-354444-        8133 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8133)
  AUTHORS   Oka T, Mazack V, Sudol M
  TITLE     Mst2 and Lats kinases regulate apoptotic function of Yes 
            kinase-associated protein (YAP).
  JOURNAL   J Biol Chem. 2008 Oct 10;283(41):27534-27546. doi: 
            10.1074/jbc.M804380200. Epub 2008 Jul 17.
  PUBMED    18640976
REFERENCE   2  (bases 1 to 8133)
  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: 
            "10.1074/jbc.M804380200"; journalName: "J Biol Chem"; date: 
            "2008-10-10- 10"; volume: "283"; issue: "41"; pages: "27534-27546"
COMMENT     Sequence Label: pEGFP C3-Lats1
FEATURES             Location/Qualifiers
     source          1..8133
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    537..584
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     609..628
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(816..1610)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(816..1610)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(927..946)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1537..1556
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1645..2002)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1659..1794
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        1811..2002
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     1959..1979
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(2004..2108)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2135..2590
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2222..2241)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2432..2453
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2597..2718
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2608..2627)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2662..2681
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6310..7020,7021..7023,7024..7026))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(6310..7020,7021..7023,7024..7026))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6352..6373)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     6701..6720
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6960..6981
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      7023..7032
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7071..7274)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(7100..7120)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        7275..7578
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(join(7753..8133,1..208))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7833..7852)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
       61 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
      121 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
      181 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
      241 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
      301 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
      361 gtaacctgag gctatggcag ggcctgccgc cccgacgttg gctgcgagcc ctgggccttc
      421 acccgaactt ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt ggccccaatg
      481 gggtctcggt ggggtatcga cagagtgcca gccctgggac cgaaccccgc gtttatgaac
      541 aaacgaccca acaccgtgcg ttttattctg tctttttatt gccgtcatag cgcgggttcc
      601 ttccggtatt gtctccttcc gtgtttcagt tagcctcccc ctagggtggg cgaagaactc
      661 cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa
      721 gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca ggttgggcgt
      781 cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc aagaaggcga
      841 tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca
      901 gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag
      961 cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc
     1021 atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg
     1081 ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga
     1141 tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc
     1201 gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca
     1261 gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc
     1321 acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg
     1381 caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc
     1441 agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg
     1501 aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc
     1561 tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat
     1621 cctcatcctg tctcttgatc gatctttgca aaagcctagg cctccaaaaa agcctcctca
     1681 ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat
     1741 tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg
     1801 agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     1861 tgctggggag cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg
     1921 catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca
     1981 cagctggttc tttccgcctc aggactcttc ctttttcaat attattgaag catttatcag
     2041 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     2101 gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc
     2161 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2221 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2281 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2341 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2401 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2461 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2521 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     2581 cttacaattt acgcgttaag atacattgat gagtttggac aaaccacaac tagaatgcag
     2641 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     2701 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     2761 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat
     2821 gatcagttat ctagaggatc tactagtcat atggatagaa gaagatctag aaacatatac
     2881 tagatcgcga tttttaatct ctgagcctgt gttttgatca tcttcatcca actgctgctc
     2941 tgagccttgt gaattaatgt attcatattc aataggcttc ggataattat atgggtagcc
     3001 attgtcatca aaaaaccttc ggaaggtaaa ttcatagaat gcatgttcag gatgctttcc
     3061 atttttatac catccattga gagtgtcatt tacattttct tcctcgttat catcactcca
     3121 taatttatca ggatcaacag gatcaaaatt tgatgtatct gttgggtgtg tgattttagg
     3181 aatgtatgaa gcagactgct gtctcaggtc actggagaaa tcaattgttt taaaaaatgg
     3241 atgagctttt atttcatcag caccattctt gcctaagcga tcttcgggtc ctcggcaaag
     3301 tttaataata agatcagaag cttcaggact gagtttagct tgtggtggaa tgtgaagaga
     3361 tgtttgccag ttgataacct tcatttgtgt ttctaatggt gtttgtgcca agaaaggagg
     3421 ttgtcccacc aacatttcaa aaagaataac accaacactc caccaatcac acaactgtgt
     3481 gtatcctgtt cgtagcaaca cttcaggtgc aatataattg ggagtcccaa ccaaagaatg
     3541 tgctagacat cgctggtgct ggcgtgcagc tctccgctct aatggcttca gtctgtctcc
     3601 acatcgacag cttgagggat ccccccattc attactgaaa tccatgctat cttgccgtgg
     3661 atggtcacca ctctgatagt acttagaatc gtgtgtccat ctgaagccag tgcagaggcc
     3721 aaagtcagtc aatttaatat gaccatcacg atcaatcaaa atattatcag gtttaatatc
     3781 tctatgaata aaacccattt tatgaacact ttcaactgca caggtaagtt ctgctatgta
     3841 gaatcgtgcc agactttctg gaaagatgcc cattctaatt aataggctca tcatatcacc
     3901 cccaggaatg tagtccatta caaagtataa attgtcctta tcttggaatg aataatatag
     3961 acgaactacc cattcattgt cagcttcagc caggatatct ctctcagcct taacatgagc
     4021 gacttgattt cgaagaagaa catctttctt tcgaagagtt tttgttgcat acaaagcctt
     4081 agtatctact tttcttgcta gacagacttc accaaatgct cctattccta gtgtctttat
     4141 cttcacaaac atagacttgt ccattttagc ccttttaaga cggatgtaat tagattcttt
     4201 ttggcaaagc atctttctca tttgatcctg ggcatcttga gataatccaa cccgcatcat
     4261 ttcattctct aattgttttt tacgatgtag acgctgctga tgagatttga gtacattttc
     4321 tacatgttgc tccataaaga atttaaatgc ttgaggagaa taactttgaa tacgagattc
     4381 ccttcgctct tcatctttct tgtttttcct aacagtaata ggtgaagttg taatctgttt
     4441 cttttcttta tccccactat caacattttc ataactcttt tcactctcat cttccttggg
     4501 caagcttggc tgatcctctt tgctaggctt actgattgac tcgtatggag gaacagatgg
     4561 gttttggtgc agcagatgtt ttgggtaggg tggtggtggt ccttgatagt ttggagcttc
     4621 agcaacaggt ggcatcacag tcacatttga agcggttccc tcaggaaaag gactgggttg
     4681 aacagtttga attggctgtg gtatccaaga agggtgtgta ggtgctaaag cagtctgtag
     4741 ctctggtttt aatacacgca tacttttcac aggctgttga ataggagctg gtgtaattgc
     4801 agtgactgtt gtagcggaag gctgagaatt agcagagtga cttgctctat ttcctaatgg
     4861 gttattaaaa gaatttgacc tcactggtat gttaggttgc catgtaggga tttcatgccc
     4921 actgctcggg gatgactggg ctggagcaga agatgactga ggccaatttg tttgcagtcc
     4981 aggtacacta atgttatata gttccatgtt atgactattt ctgtttggca ccatcataga
     5041 ctgaggaata cttccatttg tatatgacga aggagcagca gatccccctg tttgtaaagc
     5101 agaagggctt tgtccattag ctgctgtcag aggatatgga ggtggtggct gccgattcac
     5161 agtgccagca gggacaacat tttggtgtat catgaaatca gtttgtccag taccattctg
     5221 cattccaggt ctccctgatg gaaagttaaa tttgctagaa ctctgcatga tgattggttg
     5281 tctgccaaca ggaacagaac taatgcctct ctgtccttga ttaggaggat tcatggggga
     5341 agtgttgaga ggtggtggag gatagccctc ttgccatgcc ccaggtggga caggagagat
     5401 tcgggagatt acgtattcca tgtttccaga atagcgcttt gtttgagagt ttggttccca
     5461 tgaaggggga ggtggagttg tacctcttgg agggggagtc tggcctcttg gaggtggtgg
     5521 aggagtaaca ctccttactt gaggtggtgg tggggggttc actctctgtc cgttgctagg
     5581 gtgagcttga acaaatgctg atataccaga tccagacaaa ggtcttccta catctgtctg
     5641 tgagttggga ctctcagaat gataggccac actttctcct agtggcgggc catgcctctg
     5701 aggaactaag gattctttag aacctttcca gctctgtttg cggttaactg attgctgcac
     5761 attccctggt ttcatgctgg cattaatagg tctggcagct gctgcagcca tctgctctcg
     5821 tcgaggatct tggtaactca ttttactaat gaattcaatt gctgcttcta tacttctgtt
     5881 gttagttttc tgaagagctt gtataaccat atcctcatca aatccagcag cttgcaagtc
     5941 ttgaagcatt tgtggattaa cttctgaagt actccgagaa gaatttgttt catttgcaaa
     6001 tggaagcaga gagtttcgaa tttcctgcaa ggctttatga tgcgtcccaa atttgggtgg
     6061 atttctgact tgtcgaggat cttcggttga cattttactc atgttatgct cagccttagc
     6121 agcatcagat ggtttagata aattcctaag ggattcccga atttcttgta acatttgccg
     6181 gctactgaca gtatagttac tggcaggaaa ggtcttaggc ctcatttgtc tatatccttc
     6241 tggcttttca ctcctcttca ttggtaccgt cgactgcaga attcgaagct tgagctcgag
     6301 atctgagtac ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc
     6361 cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg actgggtgct
     6421 caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta
     6481 gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttcacctt
     6541 gatgccgttc ttctgcttgt cggccatgat atagacgttg tggctgttgt agttgtactc
     6601 cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg
     6661 gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc
     6721 cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc
     6781 gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg tcagggtggt
     6841 cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag
     6901 cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac
     6961 gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct
     7021 caccatggtg gcgaccggta gcgctagcgg atctgacggt tcactaaacc agctctgctt
     7081 atatagacct cccaccgtac acgcctaccg cccatttgcg tcaatggggc ggagttgtta
     7141 cgacattttg gaaagtcccg ttgattttgg tgccaaaaca aactcccatt gacgtcaatg
     7201 gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt gatgtactgc
     7261 caaaaccgca tcaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga
     7321 aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg
     7381 tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac
     7441 cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca
     7501 tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt
     7561 accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt
     7621 gattactatt aataactaat gcatggcggt aatacggtta tccacagaat caggggataa
     7681 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     7741 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7801 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7861 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7921 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7981 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     8041 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     8101 agcagccact ggtaacagga ttagcagagc gag
//
