LOCUS       sequence-119836-         989 bp    DNA     linear   UNA 15-APR-2025
DEFINITION  Standard for cell-free expression benchmarking..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 989)
  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 989)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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 TFF1
FEATURES             Location/Qualifiers
     source          1..989
                     /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"
     regulatory      67..76
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(317..341)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      565..611
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(689..705)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(689..705)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(702..724)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    713..729
                     /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(737..743,744..761,762..767))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    782..803
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(920..937)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gacgagctat acaagctgga ggtgctcttc cagggtccga caagtttgta caaaaaagtt
       61 ggcatggcca ccatggagaa caaggtgatc tgcgccctgg tcctggtgtc catgctggcc
      121 ctcggcaccc tggccgaggc ccagacagag acgtgtacag tggccccccg tgaaagacag
      181 aattgtggtt ttcctggtgt cacgccctcc cagtgtgcaa ataagggctg ctgtttcgac
      241 gacaccgttc gtggggtccc ctggtgcttc tatcctaata ccatcgacgt ccctccagaa
      301 gaggagtgtg aattttaccc aactttcttg tacaaagtgg tgtaagcggc cgccctcctc
      361 ctcctttctt gttcctttca cgtcgccttc tcggttgtag ctggcagacg acgagtctta
      421 cttttacgtg tacttctcta tagatgatgt atgatctctc tgcatgcgtg ttcgtgcatg
      481 tgtccgtgtg ttgggtacgc gtggtaccct gcaggaagga agctgagttg gctgctgcca
      541 ccgctgagca ataactagta attactagca taaccccttg gggcctctaa acgggtcttg
      601 agggggtttt ttgctgaaag gaggacagct gatgattgtc atgcttgcca tctgttttct
      661 tgcaaggtca gaggaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc
      721 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
      781 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
      841 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
      901 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
      961 gagcggtatc agctcactca aaggcggta
//
