LOCUS       sequence-66896-e         961 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 961)
  AUTHORS   Maier CJ, Maier RH, Hintner H, Bauer JW, Onder K
  TITLE     Coupled yeast 2-hybrid-mammalian 2-hybrid reading-frame-independent 
            and site-specific recombinational cloning vector system.
  JOURNAL   Assay Drug Dev Technol. 2010 Oct;8(5):625-9. doi: 
            10.1089/adt.2009.0266. Epub 2010 Jun 1.
  PUBMED    20515414
REFERENCE   2  (bases 1 to 961)
  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: 
            "10.1089/adt.2009.0266"; journalName: "Assay Drug Dev Technol"; 
            date: "2010-10"; volume: "8"; issue: "5"; pages: "625-9"
COMMENT     Sequence Label: pMAD-Gate5
FEATURES             Location/Qualifiers
     source          1..961
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        46..64
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             128..148
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    341..465
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(506..811)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(608..627)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
ORIGIN
        1 ggtgtccact cccagttcaa ttacagctct taaggctaga gtacttaata cgactcacta
       61 taggctagcc agcttgaagc aagcctcctg aaagatgaag ctactgtctt ctatcgaaca
      121 agcatgccca aaaaagaaga gaaaggtaga tgaattcccg gggatctcga cggccccccc
      181 gaccgatgtc agcctggggg acgagctcca cttagacggc gaggacgtgg cgatggcgca
      241 tgccgacgcg ctagacgatt tcgatctgga catgttgggg gacggggatt ccccgggtcc
      301 gggatcgcca gggatccgtc gacttgacgc gttgatatca accactttgt acaagaaagc
      361 tgaacgagaa acgtaaaatg atataaatat caatatatta aattagattt tgcataaaaa
      421 acagactaca taatactgta aaacacaaca tatccagtca ctatggtcga cctgcagact
      481 ggctgtgtat aagggagcct gacatttata ttccccagaa catcaggtta atggcgtttt
      541 tgatgtcatt ttcgcggtgg ctgagatcag ccacttcttc cccgataacg gagaccggca
      601 cactggccat atcggtggtc atcatgcgcc agctttcatc cccgatatgc accaccgggt
      661 aaagttcacg ggagacttta tctgacagca gacgtgcact ggccaggggg atcaccatcc
      721 gtcgcccggg cgtgtcaata atatcactct gtacatccac aaacagacga taacggctct
      781 ctcttttata ggtgtaaacc ttaaactgca tttcaccagc ccctgttctc gtcagcaaaa
      841 gagccgttca tttcaataaa ccgggcgacc tcagccatcc cttcctgatt ttccgctttc
      901 cagcgttcgg cacgcagacg acgggcttca ttctgcatgg ttgtgcttac cagaccggag
      961 a
//
