LOCUS       sequence-119813-         919 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 919)
  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 919)
  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 TCEB2
FEATURES             Location/Qualifiers
     source          1..919
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             16..39
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     protein_bind    40..64
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(419..443)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      667..713
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(791..807)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(791..807)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(804..826)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    815..831
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(839..845,846..863,864..869))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    884..905
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gacgagctat acaagctgga ggtgctcttc cagggtccga caagtttgta caaaaaagtt
       61 ggcatggacg tgttcctcag gatccggcgc cacaagacca ccatcttcac ggacgccaag
      121 gagtccagca cggtgttcga actgaagcgc atcgtcgagg gcatcctcaa gcggcctcct
      181 gacgagcagc ggctgtacaa ggatgaccaa ctcttggatg atggcaagac actgggcgag
      241 tgtggcttca ccagtcaaac agcacggcca caggccccag ccacagtggg gctggccttc
      301 cgggcagatg acacctttga ggccctgtgc atcgagccgt tttccagccc gccagagctg
      361 cccgatgtga tgaagcccca ggactcggga agcagtgcca atgaaaaagc cgtgcagtgc
      421 ccaactttct tgtacaaagt ggtgtaagcg gccgccctcc tcctcctttc ttgttccttt
      481 cacgtcgcct tctcggttgt agctggcaga cgacgagtct tacttttacg tgtacttctc
      541 tatagatgat gtatgatctc tctgcatgcg tgttcgtgca tgtgtccgtg tgttgggtac
      601 gcgtggtacc ctgcaggaag gaagctgagt tggctgctgc caccgctgag caataactag
      661 taattactag cataacccct tggggcctct aaacgggtct tgagggggtt ttttgctgaa
      721 aggaggacag ctgatgattg tcatgcttgc catctgtttt cttgcaaggt cagaggaatt
      781 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
      841 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
      901 cattaattgc gttgcgctc
//
