LOCUS       sequence-237604-        6109 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 6109)
  AUTHORS   Basu S, Totty NF, Irwin MS, Sudol M, Downward J
  TITLE     Akt phosphorylates the Yes-associated protein, YAP, to induce 
            interaction with 14-3-3 and attenuation of p73-mediated apoptosis.
  JOURNAL   Mol Cell. 2003 Jan . 11(1):11-23.
  PUBMED    12535517
REFERENCE   2  (bases 1 to 6109)
  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: "Mol Cell. 
            2003 Jan . 11(1):11-23."
COMMENT     Sequence Label: pEGFP-C3-hYAP1
FEATURES             Location/Qualifiers
     source          1..6109
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        70..427
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        70..261
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(93..113)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      278..413
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             462..1256
                     /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             462..1256
                     /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(516..535)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1126..1145
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1444..1463)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1488..1535
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1864..2452
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2353..2372
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2627..2930
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2931..3134
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3085..3105
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3173..3182
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3179..3181,3182..3184,3185..3895)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3179..3181,3182..3184,3185..3895)
                     /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(3224..3245)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3485..3504)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3832..3853
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5463..5584
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5500..5519)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5554..5573
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5591..6046)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5728..5749)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5940..5959
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(6073..6109,1..68)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
       61 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
      121 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
      181 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
      241 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
      301 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
      361 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
      421 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
      481 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
      541 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
      601 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
      661 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
      721 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
      781 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
      841 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
      901 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
      961 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     1021 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     1081 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     1141 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     1201 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     1261 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     1321 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     1381 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     1441 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     1501 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     1561 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     1621 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     1681 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     1741 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     1801 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     1861 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     1921 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     1981 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     2041 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2101 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2161 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2221 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2281 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2341 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2401 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2461 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     2521 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt
     2581 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     2641 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     2701 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     2761 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     2821 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     2881 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     2941 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     3001 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     3061 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     3121 atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat
     3181 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     3241 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     3301 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     3361 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     3421 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     3481 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     3541 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     3601 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     3661 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     3721 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     3781 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     3841 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccgg
     3901 actcagatct cgagctcaag cttcgaattg tacaggatcc ccgcatatgg atcccgggca
     3961 gcagccgccg cctcaaccgg ccccccaggg ccaagggcag ccgccttcgc agcccccgca
     4021 ggggcagggc ccgccgtccg gacccgggca accggcaccc gcggcgaccc aggcggcgcc
     4081 gcaggcaccc cccgccgggc atcagatcgt gcacgtccgc ggggactcgg agaccgacct
     4141 ggaggcgctc ttcaacgccg tcatgaaccc caagacggcc aacgtgcccc agaccgtgcc
     4201 catgaggctc cggaagctgc ccgactcctt cttcaagccg ccggagccca aatcccactc
     4261 ccgacaggcc agtactgatg caggcactgc aggagccctg actccacagc atgttcgagc
     4321 tcattcctct ccagcttctc tgcagttggg agctgtttct cctgggacac tgacccccac
     4381 tggagtagtc tctggcccag cagctacacc cacagctcag catcttcgac agtcttcttt
     4441 tgagatacct gatgatgtac ctctgccagc aggttgggag atggcaaaga catcttctgg
     4501 tcagagatac ttcttaaatc acatcgatca gacaacaaca tggcaggacc ccaggaaggc
     4561 catgctgtcc cagatgaacg tcacagcccc caccagtcca ccagtgcagc agaatatgat
     4621 gaactcggct tcagccatga accagagaat cagtcagagt gctccagtga aacagccacc
     4681 acccctggct ccccagagcc cacagggagg cgtcatgggt ggcagcaact ccaaccagca
     4741 gcaacagatg cgactgcagc aactgcagat ggagaaggag aggctgcggc tgaaacagca
     4801 agaactgctt cggcaggtga ggccacagga gttagccctg cgtagccagt taccaacact
     4861 ggagcaggat ggtgggactc aaaatccagt gtcttctccc gggatgtctc aggaattgag
     4921 aacaatgacg accaatagct cagatccttt ccttaacagt ggcacctatc actctcgaga
     4981 tgagagtaca gacagtggac taagcatgag cagctacagt gtccctcgaa ccccagatga
     5041 cttcctgaac agtgtggatg agatggatac aggtgatact atcaaccaaa gcaccctgcc
     5101 ctcacagcag aaccgtttcc cagactacct tgaagccatt cctgggacaa atgtggacct
     5161 tggaacactg gaaggagatg gaatgaacat agaaggagag gagctgatgc caagtctgca
     5221 ggaagctttg agttctgaca tccttaatga catggagtct gttttggctg ccaccaagct
     5281 agataaagaa agctttctta catggttata gagccctcag gcagactgaa ttcagatcca
     5341 ccggatctag ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     5401 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     5461 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5521 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5581 cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     5641 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     5701 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     5761 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     5821 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     5881 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     5941 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     6001 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     6061 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacatt
//
