LOCUS       sequence-119701-        1076 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cell-free expression control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1076)
  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 1076)
  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_G05 sfGFP
FEATURES             Location/Qualifiers
     source          1..1076
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    166..190
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(192..902,903..905))
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             complement(192..905)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(232..252)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     847..875
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     protein_bind    complement(908..932)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(933..956)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
ORIGIN
        1 taccacgcgt acccaacaca cggacacatg cacgaacacg catgcagaga gatcatacat
       61 catctataga gaagtacacg taaaagtaag actcgtcgtc tgccagctac aaccgagaag
      121 gcgacgtgaa aggaacaaga aaggaggagg agggcggccg cttacaccac tttgtacaag
      181 aaagctgggt ttttgtagag ctcatccatg ccatgtgtaa tcccagcagc agttacaaac
      241 tcaagaagga ccatgtggtc acgcttttcg ttgggatctt tcgaaaggac agattgtgtc
      301 gacaggtaat ggttgtctgg taaaaggaca gggccatcgc caattggagt attttgttga
      361 taatggtctg ctagttgaac ggaaccatct tcaacgttgt ggcgaatttt gaagttagct
      421 ttgattccat tcttttgttt gtctgccgtg atgtatacat tgtgtgagtt aaagttgtac
      481 tcgagtttgt gtccaagaat gtttccatct tctttaaaat caataccctt taactcgata
      541 cgattaacaa gggtatcacc ttcaaacttg acttcagcac gcgtcttgta ggtcccgtca
      601 tctttgaaag atatagtgcg ttcctgtaca taaccttcgg gcatggcact cttgaaaaag
      661 tcatgccgtt tcatgtgatc cggataacgg gaaaagcatt gaacaccata ggtcagagta
      721 gtgacaagtg ttggccacgg aacaggtagt tttccagtag tgcaaataaa tttaagggtg
      781 agttttccgt ttgtagcatc accttcaccc tctccacgga cagaaaattt gtgcccatta
      841 acatcaccat ctaattcaac aagaattggg acaactccag tgaaaagttc ttctcctttg
      901 ctcatgaagc ctgctttttt gtacaaactt gtcggaccct ggaagagcac ctccagcttg
      961 tacagctcgt ccatgccgcc ggtggagtgg cggccctcgg cccgttcgta ctgttccacg
     1021 atggtgtagt cctcgttggg ggaggtgatg tccaacttga tgttgacgtt gtaggc
//
