LOCUS       sequence-119808-         897 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 897)
  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 897)
  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 S100A1
FEATURES             Location/Qualifiers
     source          1..897
                     /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(347..371)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      595..641
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(719..735)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(719..735)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(732..754)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    743..759
                     /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(767..773,774..791,792..797))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    812..833
                     /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 ggcatgggct ctgagctgga gacggcgatg gagaccctca tcaacgtgtt ccacgcccac
      121 tcgggcaaag agggggacaa gtacaagctg agcaagaagg agctgaaaga gctgctgcag
      181 acggagctct ctggcttcct ggatgcccag aaggatgtgg atgctgtgga caaggtgatg
      241 aaggagctag acgagaatgg agacggggag gtggacttcc aggagtatgt ggtgcttgtg
      301 gctgctctca cagtggcctg taacaatttc ttctgggaga acagttgccc aactttcttg
      361 tacaaagtgg tgtaagcggc cgccctcctc ctcctttctt gttcctttca cgtcgccttc
      421 tcggttgtag ctggcagacg acgagtctta cttttacgtg tacttctcta tagatgatgt
      481 atgatctctc tgcatgcgtg ttcgtgcatg tgtccgtgtg ttgggtacgc gtggtaccct
      541 gcaggaagga agctgagttg gctgctgcca ccgctgagca ataactagta attactagca
      601 taaccccttg gggcctctaa acgggtcttg agggggtttt ttgctgaaag gaggacagct
      661 gatgattgtc atgcttgcca tctgttttct tgcaaggtca gaggaattcg taatcatggt
      721 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
      781 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
      841 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaa
//
