LOCUS       sequence-121782-         854 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Gateway destination vector for cell-free expression with N-term 
            8xHis tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 854)
  AUTHORS   Gagoski D, Mureev S, Giles N, Johnston W, Dahmer-Heath M, Skalamera 
            D, Gonda TJ, Alexandrov K
  TITLE     Gateway-compatible vectors for high-throughput protein expression in
            pro- and eukaryotic cell-free systems.
  JOURNAL   J Biotechnol. 2015 Feb 10;195:1-7. doi: 
            10.1016/j.jbiotec.2014.12.006. Epub 2014 Dec 19.
  PUBMED    25529348
REFERENCE   2  (bases 1 to 854)
  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: "J 
            Biotechnol."; date: "2015-02-10"; volume: "195:1-7. doi"; pages: " 
            10.1016/j.jbiotec.2014.12.006. Epub 2014 Dec 19"
COMMENT     Sequence Label: pCellFree_G01
FEATURES             Location/Qualifiers
     source          1..854
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             107..130
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     protein_bind    134..258
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(305..610)
                     /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(407..426)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
ORIGIN
        1 tattttattt attttatttt attttatttt attttatttt atttaaccat gacagctaat
       61 gtataaagtc tgtaaagaca ttaaacacgt aagtgaaacc atgggccacc accatcacca
      121 ccatcaccac ggcacaagtt tgtacaaaaa agctgaacga gaaacgtaaa atgatataaa
      181 tatcaatata ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca
      241 acatatccag tcactatggg gcccgtcgac ctgcagactg gctgtgtata agggagcctg
      301 acatttatat tccccagaac atcaggttaa tggcgttttt gatgtcattt tcgcggtggc
      361 tgagatcagc cacttcttcc ccgataacgg agaccggcac actggccata tcggtggtca
      421 tcatgcgcca gctttcatcc ccgatatgca ccaccgggta aagttcacgg gagactttat
      481 ctgacagcag acgtgcactg gccaggggga tcaccatccg tcgcccgggc gtgtcaataa
      541 tatcactctg tacatccaca aacagacgat aacggctctc tcttttatag gtgtaaacct
      601 taaactgcat ttcaccagcc cctgttctcg tcagcaaaag agccgttcat ttcaataaac
      661 cgggcgacct cagccatccc ttcctgattt tccgctttcc agcgttcggc acgcagacga
      721 cgggcttcat tctgcatggt tgtgcttacc agaccggaga tattgacatc atatatgcct
      781 tgagcaactg atagctgtcg ctgtcaactg tcactgtaat acgctgcttc atagcatacc
      841 tccttttttg acat
//
