LOCUS       sequence-120275-         922 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 922)
  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 922)
  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 CSNK2A1
FEATURES             Location/Qualifiers
     source          1..922
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             121..144
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             151..861
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     CDS             151..861
                     /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(190..211)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(451..470)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     798..819
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             862..885
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     protein_bind    886..910
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ttatttattt tattttattt tattttattt tattttattt tattttattt aaccatgaca
       61 gtaatgtata aagtctgtaa agacattaaa cacgtaagtg aaaccatggg cacctctggt
      121 caccaccatc accaccatca ccacggcaca agcaagggcg aggagctgtt caccggggtg
      181 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
      241 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
      301 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
      361 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
      421 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
      481 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
      541 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
      601 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
      661 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
      721 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
      781 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
      841 ggcatggacg agctatacaa gctggaggtg ctcttccagg gtccgacaag tttgtacaaa
      901 aaagttggca tctcgggaac cc
//
