LOCUS       sequence-121136-         985 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Gateway destination vector for cell-free expression with N-term 
            8xHis eGFP tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 985)
  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 985)
  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_G03
FEATURES             Location/Qualifiers
     source          1..985
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             16..39
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     protein_bind    40..164
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(211..516)
                     /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(313..332)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
ORIGIN
        1 gacgagctat acaagctgga ggtgctcttc cagggtccga caagtttgta caaaaaagct
       61 gaacgagaaa cgtaaaatga tataaatatc aatatattaa attagatttt gcataaaaaa
      121 cagactacat aatactgtaa aacacaacat atccagtcac tatggggccc gtcgacctgc
      181 agactggctg tgtataaggg agcctgacat ttatattccc cagaacatca ggttaatggc
      241 gtttttgatg tcattttcgc ggtggctgag atcagccact tcttccccga taacggagac
      301 cggcacactg gccatatcgg tggtcatcat gcgccagctt tcatccccga tatgcaccac
      361 cgggtaaagt tcacgggaga ctttatctga cagcagacgt gcactggcca gggggatcac
      421 catccgtcgc ccgggcgtgt caataatatc actctgtaca tccacaaaca gacgataacg
      481 gctctctctt ttataggtgt aaaccttaaa ctgcatttca ccagcccctg ttctcgtcag
      541 caaaagagcc gttcatttca ataaaccggg cgacctcagc catcccttcc tgattttccg
      601 ctttccagcg ttcggcacgc agacgacggg cttcattctg catggttgtg cttaccagac
      661 cggagatatt gacatcatat atgccttgag caactgatag ctgtcgctgt caactgtcac
      721 tgtaatacgc tgcttcatag catacctctt tttgacatac ttcgggtata catatcagta
      781 tatattctta taccgcaaaa atcagcgcgc aaatacgcat actgttatct ggcttttagt
      841 aagccggatc cacgcgttta cgccccgccc tgccactcat cgcagtactg ttgtaattca
      901 ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc
      961 ggcatcagca ccttgtcgcc ttgcg
//
