LOCUS       sequence-8576-ex        3024 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for N-term eGFP fusion, frame C3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3024)
  AUTHORS   Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO, 
            Campisi J, Yaswen P, Cooper PK, Kaufman PD.
  TITLE     A versatile viral system for expression and depletion of proteins in
            mammalian cells.
  JOURNAL   PLoS One. 2009 Aug 6;4(8):e6529.
  PUBMED    19657394
REFERENCE   2  (bases 1 to 3024)
  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: "PLoS One.";
            date: "2009-08-6"; volume: "4(8)"; pages: "e6529"
COMMENT     Sequence Label: pENTR4-GFP-C3 (w393-1)
FEATURES             Location/Qualifiers
     source          1..3024
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(463..465,466..468,469..1179)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(463..465,466..468,469..1179)
                     /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(508..529)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(769..788)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1116..1137
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1185..1241
                     /label=MCS
                     /note="multiple cloning site"
     protein_bind    complement(1253..1352)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1464..1483)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1475..2284
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1542..1561)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2374..2962
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2863..2882
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggctcca ccatggtgag caagggcgag
      481 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      541 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
      601 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
      661 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
      721 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
      781 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
      841 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
      901 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
      961 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     1021 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     1081 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1141 gccgccggga tcactctcgg catggacgag ctgtacaagt actcagatct cgagctcaag
     1201 cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agacccagct
     1261 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
     1321 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgcagctct ggcccgtgtc
     1381 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     1441 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     1501 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     1561 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     1621 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     1681 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     1741 tggtgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     1801 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     1861 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     1921 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     1981 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     2041 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     2101 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     2161 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     2221 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     2281 ctaatcagaa ttggttaatt ggttgtaaca ttattcagat tgggccccgt tccactgagc
     2341 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2401 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2461 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2521 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2581 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2641 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2701 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2761 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2821 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2881 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2941 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3001 ttgctggcct tttgctcaca tgtt
//
