LOCUS       sequence-120868-        1010 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 1010)
  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 1010)
  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 AKT1
FEATURES             Location/Qualifiers
     source          1..1010
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(21..44)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(187..206)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(188..206)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        238..342
                     /gene="bla"
                     /label=AmpR promoter
     CDS             527..905
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     complement(561..580)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ctcccgccct tgcttgtgcc gtggtgatgg tggtgatggt ggtgaccaga ggtgcccatg
       61 gtttcactta cgtgtttaat gtctttacag actttataca ttactgtcat ggttaaataa
      121 aataaaataa aataaaataa aataaaataa aataaaataa aataaaataa aataaactta
      181 agatgtccct atagtgagtc gtattagacg tcaggtggca cttttcgggg aaatgtgcgc
      241 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
      301 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
      361 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
      421 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
      481 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
      541 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
      601 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
      661 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
      721 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
      781 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
      841 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
      901 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
      961 gactgggatg gaggcggata aagttgcagg accacttctg cgctcggccc
//
