LOCUS       sequence-119995-         980 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 980)
  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 980)
  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 ISG15
FEATURES             Location/Qualifiers
     source          1..980
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             12..35
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     protein_bind    36..60
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(556..580)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      804..850
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(928..944)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(928..944)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(941..963)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    952..968
                     /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)."
ORIGIN
        1 gagagcttca gctggaggtg ctcttccagg gtccgacaag tttgtacaaa aaagttggca
       61 tgggctggga cctgacggtg aagatgctgg cgggcaacga attccaggtg tccctgagca
      121 gctccatgtc ggtgtcagag ctgaaggcgc agatcaccca gaagatcggc gtgcacgcct
      181 tccagcagcg tctggctgtc cacccgagcg gtgtggcgct gcaggacagg gtcccccttg
      241 ccagccaggg cctgggcccc ggcagcacgg tcctgctggt ggtggacaaa tgcgacgaac
      301 ctctgaacat cctggtgagg aataacaagg gccgcagcag cacctacgag gtgcggctga
      361 cgcagaccgt ggcccacctg aagcagcaag tgagcgggct ggagggtgtg caggacgacc
      421 tgttctggct gaccttcgag gggaagcccc tggaggacca gctcccgctg ggggagtacg
      481 gcctcaagcc cctgagcacc gtgttcatga atctgcgcct gcggggaggc ggcacagagc
      541 ctggcgggcg gagctaccca actttcttgt acaaagtggt gtaagcggcc gccctcctcc
      601 tcctttcttg ttcctttcac gtcgccttct cggttgtagc tggcagacga cgagtcttac
      661 ttttacgtgt acttctctat agatgatgta tgatctctct gcatgcgtgt tcgtgcatgt
      721 gtccgtgtgt tgggtacgcg tggtaccctg caggaaggaa gctgagttgg ctgctgccac
      781 cgctgagcaa taactagtaa ttactagcat aaccccttgg ggcctctaaa cgggtcttga
      841 gggggttttt tgctgaaagg aggacagctg atgattgtca tgcttgccat ctgttttctt
      901 gcaaggtcag aggaattcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
      961 gctcacaatt ccacacaaca
//
