LOCUS       sequence-8638-ex        3892 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector, Firefly Luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3892)
  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 3892)
  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: pENTR-LUC (w158-1)
FEATURES             Location/Qualifiers
     source          1..3892
                     /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             463..2115
                     /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(529..547)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     protein_bind    complement(2122..2221)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2333..2352)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2344..3153
                     /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(2411..2430)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3243..3831
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3732..3751
                     /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 ccatggaaga cgccaaaaac
      481 ataaagaaag gcccggcgcc attctatccg ctggaagatg gaaccgctgg agagcaactg
      541 cataaggcta tgaagagata cgccctggtt cctggaacaa ttgcttttac agatgcacat
      601 atcgaggtgg acatcactta cgctgagtac ttcgaaatgt ccgttcggtt ggcagaagct
      661 atgaaacgat atgggctgaa tacaaatcac agaatcgtcg tatgcagtga aaactctctt
      721 caattcttta tgccggtgtt gggcgcgtta tttatcggag ttgcagttgc gcccgcgaac
      781 gacatttata atgaacgtga attgctcaac agtatgggca tttcgcagcc taccgtggtg
      841 ttcgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc aaaaaaagct cccaatcatc
      901 caaaaaatta ttatcatgga ttctaaaacg gattaccagg gatttcagtc gatgtacacg
      961 ttcgtcacat ctcatctacc tcccggtttt aatgaatacg attttgtgcc agagtccttc
     1021 gatagggaca agacaattgc actgatcatg aactcctctg gatctactgg tctgcctaaa
     1081 ggtgtcgctc tgcctcatag aactgcctgc gtgagattct cgcatgccag agatcctatt
     1141 tttggcaatc aaatcattcc ggatactgcg attttaagtg ttgttccatt ccatcacggt
     1201 tttggaatgt ttactacact cggatatttg atatgtggat ttcgagtcgt cttaatgtat
     1261 agatttgaag aagagctgtt tctgaggagc cttcaggatt acaagattca aagtgcgctg
     1321 ctggtgccaa ccctattctc cttcttcgcc aaaagcactc tgattgacaa atacgattta
     1381 tctaatttac acgaaattgc ttctggtggc gctcccctct ctaaggaagt cggggaagcg
     1441 gttgccaaga ggttccatct gccaggtatc aggcaaggat atgggctcac tgagactaca
     1501 tcagctattc tgattacacc cgagggggat gataaaccgg gcgcggtcgg taaagttgtt
     1561 ccattttttg aagcgaaggt tgtggatctg gataccggga aaacgctggg cgttaatcaa
     1621 agaggcgaac tgtgtgtgag aggtcctatg attatgtccg gttatgtaaa caatccggaa
     1681 gcgaccaacg ccttgattga caaggatgga tggctacatt ctggagacat agcttactgg
     1741 gacgaagacg aacacttctt catcgttgac cgcctgaagt ctctgattaa gtacaaaggc
     1801 tatcaggtgg ctcccgctga attggaatcc atcttgctcc aacaccccaa catcttcgac
     1861 gcaggtgtcg caggtcttcc cgacgatgac gccggtgaac ttcccgccgc cgttgttgtt
     1921 ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg attacgtcgc cagtcaagta
     1981 acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg acgaagtacc gaaaggtctt
     2041 accggaaaac tcgacgcaag aaaaatcaga gagatcctca taaaggccaa gaagggcgga
     2101 aagatcgccg tgtaattcta gacccagctt tcttgtacaa agttggcatt ataagaaagc
     2161 attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt
     2221 gccatccagc tgcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat
     2281 aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
     2341 gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct
     2401 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     2461 cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     2521 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     2581 ccgaccatca agcattttat ccgtactcct ggtgatgcat ggttactcac cactgcgatc
     2641 cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     2701 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     2761 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     2821 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     2881 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     2941 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     3001 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     3061 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     3121 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacat
     3181 tattcagatt gggccccgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3241 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3301 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3361 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     3421 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3481 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3541 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3601 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3661 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3721 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3781 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3841 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
