LOCUS       sequence-120834-        1080 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 1080)
  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 1080)
  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 HDAC1
FEATURES             Location/Qualifiers
     source          1..1080
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             119..142
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             149..859
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     CDS             149..859
                     /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(188..209)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(449..468)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     796..817
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             860..883
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     protein_bind    884..908
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 atttatttta ttttatttta ttttatttta ttttatttta ttttatttaa ccatgacagt
       61 aatgtataaa gtctgtaaag acattaaaca cgtaagtgaa accatgggca cctctggtca
      121 ccaccatcac caccatcacc acggcacaag caagggcgag gagctgttca ccggggtggt
      181 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
      241 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
      301 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
      361 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
      421 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
      481 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
      541 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
      601 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
      661 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
      721 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
      781 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
      841 catggacgag ctatacaagc tggaggtgct cttccagggt ccgacaagtt tgtacaaaaa
      901 agttggcatg gcgcagacgc agggcacccg gaggaaagtc tgttactact acgacgggga
      961 tgttggaaat tactattatg gacaaggcca cccaatgaag cctcaccgaa tccgcatgac
     1021 tcataatttg ctgctcaact atggttctct accgaaaaat ggaaatctat cgccctccaa
//
