LOCUS       sequence-119829-         992 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 992)
  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 992)
  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 E-Cad
FEATURES             Location/Qualifiers
     source          1..992
                     /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(512..536)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      760..806
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(884..900)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(884..900)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(897..919)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    908..924
                     /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(932..938,939..956,957..962))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 gacgagctat acaagctgga ggtgctcttc cagggtccga caagtttgta caaaaaagca
       61 ggcttcgcgg tggtcaaaga gcccttactg cccccagagg atgacacccg ggacaacgtt
      121 tattactatg atgaagaagg aggcggagaa gaggaccagg actttgactt gagccagctg
      181 cacaggggcc tggacgctcg gcctgaagtg actcgtaacg acgttgcacc aaccctcatg
      241 agtgtccccc ggtatcttcc ccgccctgcc aatcccgatg aaattggaaa ttttattgat
      301 gaaaatctga aagcggctga tactgacccc acagccccgc cttatgattc tctgctcgtg
      361 tttgactatg aaggaagcgg ttccgaagct gctagtctga gctccctgaa ctcctcagag
      421 tcagacaaag accaggacta tgactacttg aacgaatggg gcaatcgctt caagaagctg
      481 gctgacatgt acggaggcgg cgaggacgac aacccagctt tcttgtacaa agtggtgtaa
      541 gcggccgccc tcctcctcct ttcttgttcc tttcacgtcg ccttctcggt tgtagctggc
      601 agacgacgag tcttactttt acgtgtactt ctctatagat gatgtatgat ctctctgcat
      661 gcgtgttcgt gcatgtgtcc gtgtgttggg tacgcgtggt accctgcagg aaggaagctg
      721 agttggctgc tgccaccgct gagcaataac tagtaattac tagcataacc ccttggggcc
      781 tctaaacggg tcttgagggg gttttttgct gaaaggagga cagctgatga ttgtcatgct
      841 tgccatctgt tttcttgcaa ggtcagagga attcgtaatc atggtcatag ctgtttcctg
      901 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
      961 aagcctgggg gtgcctaatg agtgagctaa ct
//
