LOCUS       sequence-8623-ex        4262 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector, Degron N-term Luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4262)
  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 4262)
  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: pL106P-LUC-ENTR1A(w451-1)
FEATURES             Location/Qualifiers
     source          1..4262
                     /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"
     regulatory      488..497
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(494..496,497..817)
                     /codon_start=1
                     /product="destabilization domain that can be stabilized by 
                     Shield1 in the ProteoTuner(TM) system"
                     /label=DD
                     /note="L106P mutant of FKBP12"
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKPE"
     CDS             824..2476
                     /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
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(890..908)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     protein_bind    complement(2491..2590)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2702..2721)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2713..3522
                     /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
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2780..2799)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3612..4200
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4101..4120
                     /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 gcaggcttta aaggaaccaa ttcagtcgac
      481 tggatccgcc accatgggag tgcaggtgga aaccatctcc ccaggagacg ggcgcacctt
      541 ccccaagcgc ggccagacct gtgtggtgca ctacaccggg atgcttgaag atggaaagaa
      601 agtcgattcc tcccgggaca gaaacaagcc ctttaagttt atgctaggca agcaggaggt
      661 gatccgaggc tgggaagaag gggttgccca gatgagtgtg ggtcagagag ccaaactgac
      721 tatatctcca gattatgcct atggtgccac tgggcaccca ggcatcatcc caccacatgc
      781 cactctcgtc ttcgatgtgg agcttctaaa accggaagaa ttcatggaag acgccaaaaa
      841 cataaagaaa ggcccggcgc cattctatcc gctggaagat ggaaccgctg gagagcaact
      901 gcataaggct atgaagagat acgccctggt tcctggaaca attgctttta cagatgcaca
      961 tatcgaggtg gacatcactt acgctgagta cttcgaaatg tccgttcggt tggcagaagc
     1021 tatgaaacga tatgggctga atacaaatca cagaatcgtc gtatgcagtg aaaactctct
     1081 tcaattcttt atgccggtgt tgggcgcgtt atttatcgga gttgcagttg cgcccgcgaa
     1141 cgacatttat aatgaacgtg aattgctcaa cagtatgggc atttcgcagc ctaccgtggt
     1201 gttcgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg caaaaaaagc tcccaatcat
     1261 ccaaaaaatt attatcatgg attctaaaac ggattaccag ggatttcagt cgatgtacac
     1321 gttcgtcaca tctcatctac ctcccggttt taatgaatac gattttgtgc cagagtcctt
     1381 cgatagggac aagacaattg cactgataat gaactcctct ggatctactg gtctgcctaa
     1441 aggtgtcgct ctgcctcata gaactgcctg cgtgagattc tcgcatgcca gagatcctat
     1501 ttttggcaat caaatcattc cggatactgc gattttaagt gttgttccat tccatcacgg
     1561 ttttggaatg tttactacac tcggatattt gatatgtgga tttcgagtcg tcttaatgta
     1621 tagatttgaa gaagagctgt ttctgaggag ccttcaggat tacaagattc aaagtgcgct
     1681 gctggtgcca accctattct ccttcttcgc caaaagcact ctgattgaca aatacgattt
     1741 atctaattta cacgaaattg cttctggtgg cgctcccctc tctaaggaag tcggggaagc
     1801 ggttgccaag aggttccatc tgccaggtat caggcaagga tatgggctca ctgagactac
     1861 atcagctatt ctgattacac ccgaggggga tgataaaccg ggcgcggtcg gtaaagttgt
     1921 tccatttttt gaagcgaagg ttgtggatct ggataccggg aaaacgctgg gcgttaatca
     1981 aagaggcgaa ctgtgtgtga gaggtcctat gattatgtcc ggttatgtaa acaatccgga
     2041 agcgaccaac gccttgattg acaaggatgg atggctacat tctggagaca tagcttactg
     2101 ggacgaagac gaacacttct tcatcgttga ccgcctgaag tctctgatta agtacaaagg
     2161 ctatcaggtg gctcccgctg aattggaatc catcttgctc caacacccca acatcttcga
     2221 cgcaggtgtc gcaggtcttc ccgacgatga cgccggtgaa cttcccgccg ccgttgttgt
     2281 tttggagcac ggaaagacga tgacggaaaa agagatcgtg gattacgtcg ccagtcaagt
     2341 aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg gacgaagtac cgaaaggtct
     2401 taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc ataaaggcca agaagggcgg
     2461 aaagtccaaa ttgtaactcg agatatctag acccagcttt cttgtacaaa gttggcatta
     2521 taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa
     2581 tcattatttg ccatccagct gcagctctgg cccgtgtctc aaaatctctg atgttacatt
     2641 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     2701 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac
     2761 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     2821 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     2881 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     2941 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     3001 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     3061 aatattgttg atgcgctggc agtgtccctg cgccggttgc attcgattcc tgtttgtaat
     3121 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     3181 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     3241 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     3301 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     3361 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     3421 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     3481 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
     3541 ttgtaacatt attcagattg ggccccgttc cactgagcgt cagacccggt agaaaagatc
     3601 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3661 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3721 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     3781 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3841 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3901 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3961 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4021 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4081 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4141 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4201 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4261 tt
//
