LOCUS       sequence-8931-ex        4016 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector, V5-Luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4016)
  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 4016)
  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-V5-LUC (w468-1)
FEATURES             Location/Qualifiers
     source          1..4016
                     /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             469..510
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     regulatory      580..589
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             586..2238
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(652..670)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     protein_bind    complement(2245..2344)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2456..2475)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2467..3276
                     /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(2534..2553)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3366..3954
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3855..3874
                     /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 ccatggaagg taagcctatc
      481 cctaaccctc tcctcggtct cgattctacg ggatccatgg gaaccaattc agtccctagg
      541 cttttgcaaa aagcttggca ttccggtact gttggtaaag ccaccatgga agacgccaaa
      601 aacataaaga aaggcccggc gccattctat ccgctggaag atggaaccgc tggagagcaa
      661 ctgcataagg ctatgaagag atacgccctg gttcctggaa caattgcttt tacagatgca
      721 catatcgagg tggacatcac ttacgctgag tacttcgaaa tgtccgttcg gttggcagaa
      781 gctatgaaac gatatgggct gaatacaaat cacagaatcg tcgtatgcag tgaaaactct
      841 cttcaattct ttatgccggt gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg
      901 aacgacattt ataatgaacg tgaattgctc aacagtatgg gcatttcgca gcctaccgtg
      961 gtgttcgttt ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa gctcccaatc
     1021 atccaaaaaa ttattatcat ggattctaaa acggattacc agggatttca gtcgatgtac
     1081 acgttcgtca catctcatct acctcccggt tttaatgaat acgattttgt gccagagtcc
     1141 ttcgataggg acaagacaat tgcactgatc atgaactcct ctggatctac tggtctgcct
     1201 aaaggtgtcg ctctgcctca tagaactgcc tgcgtgagat tctcgcatgc cagagatcct
     1261 atttttggca atcaaatcat tccggatact gcgattttaa gtgttgttcc attccatcac
     1321 ggttttggaa tgtttactac actcggatat ttgatatgtg gatttcgagt cgtcttaatg
     1381 tatagatttg aagaagagct gtttctgagg agccttcagg attacaagat tcaaagtgcg
     1441 ctgctggtgc caaccctatt ctccttcttc gccaaaagca ctctgattga caaatacgat
     1501 ttatctaatt tacacgaaat tgcttctggt ggcgctcccc tctctaagga agtcggggaa
     1561 gcggttgcca agaggttcca tctgccaggt atcaggcaag gatatgggct cactgagact
     1621 acatcagcta ttctgattac acccgagggg gatgataaac cgggcgcggt cggtaaagtt
     1681 gttccatttt ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct gggcgttaat
     1741 caaagaggcg aactgtgtgt gagaggtcct atgattatgt ccggttatgt aaacaatccg
     1801 gaagcgacca acgccttgat tgacaaggat ggatggctac attctggaga catagcttac
     1861 tgggacgaag acgaacactt cttcatcgtt gaccgcctga agtctctgat taagtacaaa
     1921 ggctatcagg tggctcccgc tgaattggaa tccatcttgc tccaacaccc caacatcttc
     1981 gacgcaggtg tcgcaggtct tcccgacgat gacgccggtg aacttcccgc cgccgttgtt
     2041 gttttggagc acggaaagac gatgacggaa aaagagatcg tggattacgt cgccagtcaa
     2101 gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt accgaaaggt
     2161 cttaccggaa aactcgacgc aagaaaaatc agagagatcc tcataaaggc caagaagggc
     2221 ggaaagatcg ccgtgtaatt ctagacccag ctttcttgta caaagttggc attataagaa
     2281 agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta
     2341 tttgccatcc agctgcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
     2401 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
     2461 ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat
     2521 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     2581 tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     2641 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     2701 cttccgacca tcaagcattt tatccgtact cctggtgatg catggttact caccactgcg
     2761 atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     2821 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     2881 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     2941 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     3001 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     3061 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     3121 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     3181 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     3241 ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa
     3301 cattattcag attgggcccc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3361 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3421 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3481 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3541 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3601 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3661 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3721 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3781 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3841 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3901 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3961 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtt
//
