LOCUS       sequence-121785-         987 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Gateway destination vector for cell-free expression with N-term 
            8xHis mCherry tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 987)
  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 987)
  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_G05
FEATURES             Location/Qualifiers
     source          1..987
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             108..131
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             138..839
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="SKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQ
                     TAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFED
                     GGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGE
                     IKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAEG
                     RHSTGGMDELYK"
     primer_bind     complement(277..295)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     545..564
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             840..863
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     protein_bind    864..983
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 tttattttat tttattttat tttattttat tttatttaac catgacagta atgtataaag
       61 tctgtaaaga cattaaacac gtaagtgaaa ccatgggcac ctctggtcac caccatcacc
      121 accatcacca cggcacaagc aagggcgagg aggataacat ggccatcatc aaggagttca
      181 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
      241 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
      301 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct
      361 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
      421 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
      481 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
      541 acggccccgt aatgcagaag aagactatgg gctgggaggc ctcctccgag cggatgtacc
      601 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
      661 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
      721 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
      781 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagc
      841 tggaggtgct cttccagggt ccgacaagtt tgtacaaaaa agctgaacga gaaacgtaaa
      901 atgatataaa tatcaatata ttaaattaga ttttgcataa aaaacagact acataatact
      961 gtaaaacaca acatatccag tcactat
//
