LOCUS       sequence-254273-        6235 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 6235)
  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 6235)
  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-hYAP2
FEATURES             Location/Qualifiers
     source          1..6235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        54..357
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        358..561
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     512..532
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      600..609
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(606..608,609..611,612..1322)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(606..608,609..611,612..1322)
                     /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(651..672)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(912..931)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1259..1280
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1328..1384
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3016..3137
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3053..3072)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3107..3126
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3144..3599)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3281..3302)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3493..3512
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3626..3730
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3732..4089
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3732..3923
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3755..3775)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3940..4075
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4124..4918
                     /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             4124..4918
                     /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(4178..4197)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4788..4807
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5106..5125)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5150..5197
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5526..6114
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6015..6034
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
       61 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      121 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      181 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      241 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      301 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      361 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      421 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      481 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      541 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg
      601 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
      661 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
      721 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
      781 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
      841 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
      901 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
      961 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1021 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     1081 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     1141 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1201 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     1261 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1321 agtactcaga tctcgagctc aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg
     1381 atcccgggca gcagccgccg cctcaaccgg ccccccaggg ccaagggcag ccgccttcgc
     1441 agcccccgca ggggcagggc ccgccgtccg gacccgggca accggcaccc gcggcgaccc
     1501 aggcggcgcc gcaggcaccc cccgccgggc atcagatcgt gcacgtccgc ggggactcgg
     1561 agaccgacct ggaggcgctc ttcaacgccg tcatgaaccc caagacggcc aacgtgcccc
     1621 agaccgtgcc catgaggctc cggaagctgc ccgactcctt cttcaagccg ccggagccca
     1681 aatcccactc ccgacaggcc agtactgatg caggcactgc aggagccctg actccacagc
     1741 atgttcgagc tcattcctct ccagcttctc tgcagttggg agctgtttct cctgggacac
     1801 tgacccccac tggagtagtc tctggcccag cagctacacc cacagctcag catcttcgac
     1861 agtcttcttt tgagatacct gatgatgtac ctctgccagc aggttgggag atggcaaaga
     1921 catcttctgg tcagagatac ttcttaaatc acatcgatca gacaacaaca tggcaggacc
     1981 ccaggaaggc catgctgtcc cagatgaacg tcacagcccc caccagtcca ccagtgcagc
     2041 agaatatgat gaactcggct tcaggtcctc ttcctgatgg atgggaacaa gccatgactc
     2101 aggatggaga aatttactat ataaaccata agaacaagac cacctcttgg ctagacccaa
     2161 ggcttgaccc tcgttttgcc atgaaccaga gaatcagtca gagtgctcca gtgaaacagc
     2221 caccacccct ggctccccag agcccacagg gaggcgtcat gggtggcagc aactccaacc
     2281 agcagcaaca gatgcgactg cagcaactgc agatggagaa ggagaggctg cggctgaaac
     2341 agcaagaact gcttcggcag gagttagccc tgcgtagcca gttaccaaca ctggagcagg
     2401 atggtgggac tcaaaatcca gtgtcttctc ccgggatgtc tcaggaattg agaacaatga
     2461 cgaccaatag ctcagatcct ttccttaaca gtggcaccta tcactctcga gatgagagta
     2521 cagacagtgg actaagcatg agcagctaca gtgtccctcg aaccccagat gacttcctga
     2581 acagtgtgga tgagatggat acaggtgata ctatcaacca aagcaccctg ccctcacagc
     2641 agaaccgttt cccagactac cttgaagcca ttcctgggac aaatgtggac cttggaacac
     2701 tggaaggaga tggaatgaac atagaaggag aggagctgat gccaagtctg caggaagctt
     2761 tgagttctga catccttaat gacatggagt ctgttttggc tgccaccaag ctagataaag
     2821 aaagctttct tacatggtta tagagccctc aggcagactg aattctgcag atatccagca
     2881 cagtggcggc cgctcgagtc tagataactg atcataatca gccataccac atttgtagag
     2941 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     3001 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3061 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3121 ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     3181 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     3241 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     3301 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     3361 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     3421 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     3481 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     3541 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     3601 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3661 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3721 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     3781 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3841 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3901 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3961 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4021 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4081 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4141 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4201 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4261 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     4321 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4381 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4441 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4501 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4561 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4621 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     4681 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4741 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4801 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4861 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     4921 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     4981 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5041 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     5101 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     5161 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     5221 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     5281 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     5341 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     5401 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5461 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5521 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5581 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5641 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5701 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5761 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5821 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5881 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5941 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6001 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6061 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6121 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6181 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta gttat
//
