LOCUS       sequence-72752-e        1056 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Targets beta-Tubulin for Cre-lox mediated knockout in T. brucei, 
            Puromycin selectable.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1056)
  AUTHORS   Kim HS, Li Z, Boothroyd C, Cross GA
  TITLE     Strategies to construct null and conditional null Trypanosoma brucei
            mutants using Cre-recombinase and loxP.
  JOURNAL   Mol Biochem Parasitol. 2013 Aug 13;191(1):16-19. doi: 
            10.1016/j.molbiopara.2013.08.001.
  PUBMED    23954366
REFERENCE   2  (bases 1 to 1056)
  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"; doi: 
            "10.1016/j.molbiopara.2013.08"; journalName: "Mol Biochem 
            Parasitol"; date: "2013-08-13- 13"; volume: "191"; issue: "1"; 
            pages: "16-19"
COMMENT     Sequence Label: pHJ33
FEATURES             Location/Qualifiers
     source          1..1056
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    222..255
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(292..888)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(372..392)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     869..888
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatctgg ctagcttgca
       61 tgcctgcaag ggggctcgaa tccccccatt ttcttctttt acatcatccc cccttcctaa
      121 ccaacctgca ggcgcacctc cctgctgtgc catcagatta ctccgggaac ctttccacgg
      181 aaaagacacg agacaaggga aaagccaaat gggcaggatc cataacttcg tataatgtat
      241 gctatacgaa gttatgaatt ctcagttagc ctcccccatc tcccggctag aggcaccggg
      301 cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg cggtgacggt
      361 gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca ggaaggcggg
      421 caccccggcg cgctcggccg cctccactcc ggggagcacg acggcgctgc ccagaccctt
      481 gccctggtgg tcgggcgaga caccgacggt ggccaggaac cacgcgggct ccttgggccg
      541 gtgcggcgcc aggaggcctt ccatctgttg ctgcgcggcc agccgggaac cgctcaactc
      601 ggccatgcgc gggccgatct cggcgaacac cgcccccgct tcgacgctct ccggcgtggt
      661 ccagaccgcc actgctgcgc cgtcgtccgc gacccacacc ttgccgatgt cgagcccgac
      721 gcgcgtgagg aagagttctt gcagctcggt gacccgctcg atgtggcggt ctggatcgac
      781 ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta cggccctggg
      841 gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac tcggtcatgg tagatctgta
      901 atgtcagtgc gagcgatcaa gaatgtaaca caaaggaata ccaagtttgt gaattttatt
      961 ttttggtgta attgaagtct tcaagttgaa aaaaatgggg aaaaggaaaa aaaactcgaa
     1021 ggcgcgtgca gcggccggcc gacgagaacg tgtacc
//
