LOCUS       sequence-8589-ex        2606 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for Degron, N-term, frame A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2606)
  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 2606)
  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-FKBP12DD N-term A (B3-1)
FEATURES             Location/Qualifiers
     source          1..2606
                     /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..786)
                     /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"
     protein_bind    complement(835..934)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1046..1065)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1057..1866
                     /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(1124..1143)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1956..2544
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2445..2464
                     /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 ccatgggagt gcaggtggaa
      481 accatctccc caggagacgg gcgcaccttc cccaagcgcg gccagacctg tgtggtgcac
      541 tacaccggga tgcttgaaga tggaaagaaa gtcgattcct cccgggacag aaacaagccc
      601 tttaagttta tgctaggcaa gcaggaggtg atccgaggct gggaagaagg ggttgcccag
      661 atgagtgtgg gtcagagagc caaactgact atatctccag attatgccta tggtgccact
      721 gggcacccag gcatcatccc accacatgcc actctcgtct tcgatgtgga gcttctaaaa
      781 ccggaagtcg actggatccg gtaccgaatt cgcggccgca ctcgagatat ctagacccag
      841 ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac
      901 aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgcagct ctggcccgtg
      961 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa
     1021 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
     1081 tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     1141 gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca
     1201 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     1261 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     1321 cctggtgatg catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta
     1381 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     1441 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
     1501 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     1561 aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg
     1621 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     1681 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     1741 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     1801 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     1861 ttctaatcag aattggttaa ttggttgtaa cattattcag attgggcccc gttccactga
     1921 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1981 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2041 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     2101 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     2161 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2221 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2281 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2341 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2401 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     2461 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     2521 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     2581 ttttgctggc cttttgctca catgtt
//
