LOCUS       sequence-122196-         916 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Standard for cell-free expression benchmarking..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 916)
  AUTHORS   Gagoski D, Polinkovsky ME, Mureev S, Kunert A, Johnston W, Gambin Y,
            Alexandrov K
  TITLE     Performance benchmarking of four cell-free protein expression 
            systems.
  JOURNAL   Biotechnol Bioeng. 2015 Aug 24. doi: 10.1002/bit.25814.
  PUBMED    26301602
REFERENCE   2  (bases 1 to 916)
  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: "Biotechnol 
            Bioeng. 2015 Aug 24. doi: 10.1002/bit.25814."
COMMENT     Sequence Label: pCellFree_G03 TP53
FEATURES             Location/Qualifiers
     source          1..916
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             123..146
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             153..863
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     CDS             153..863
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     primer_bind     complement(192..213)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(453..472)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     800..821
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             864..887
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     protein_bind    888..912
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 tttattttat tttattttat tttattttat tttattttat tttattttat ttaaccatga
       61 cagtaatgta taaagtctgt aaagacatta aacacgtaag tgaaaccatg ggcacctctg
      121 gtcaccacca tcaccaccat caccacggca caagcaaggg cgaggagctg ttcaccgggg
      181 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
      241 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
      301 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
      361 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
      421 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
      481 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
      541 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
      601 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
      661 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
      721 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
      781 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
      841 tcggcatgga cgagctatac aagctggagg tgctcttcca gggtccgaca agtttgtaca
      901 aaaaagttgg catgga
//
