LOCUS       sequence-8588-ex        2624 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for Degron, C-term.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2624)
  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 2624)
  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: pENTR1A-FKBP12DD C-term (w350-2)
FEATURES             Location/Qualifiers
     source          1..2624
                     /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             524..844
                     /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="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KPE"
     protein_bind    complement(853..952)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1064..1083)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1075..1884
                     /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(1142..1161)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1974..2562
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2463..2482
                     /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 tggatccggt accgaattcg cggccgcact cgagatatct agaggagtgc aggtggaaac
      541 catctcccca ggagacgggc gcaccttccc caagcgcggc cagacctgtg tggtgcacta
      601 caccgggatg cttgaagatg gaaagaaagt cgattcctcc cgggacagaa acaagccctt
      661 taagtttatg ctaggcaagc aggaggtgat ccgaggctgg gaagaagggg ttgcccagat
      721 gagtgtgggt cagagagcca aactgactat atctccagat tatgcctatg gtgccactgg
      781 gcacccaggc atcatcccac cacatgccac tctcgtcttc gatgtggagc ttctaaaacc
      841 ggaatgagct agacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat
      901 caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag
      961 ctgcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
     1021 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     1081 catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat
     1141 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat
     1201 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     1261 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     1321 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa
     1381 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     1441 gcagtgtccc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     1501 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     1561 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     1621 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     1681 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     1741 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     1801 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     1861 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ttattcagat
     1921 tgggccccgt tccactgagc gtcagacccg gtagaaaaga tcaaaggatc ttcttgagat
     1981 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2041 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2101 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     2161 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2221 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2281 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2341 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2401 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2461 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2521 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2581 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
