LOCUS       sequence-121137-         924 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Gateway destination vector for cell-free expression with C-term eGFP
            8xHis tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 924)
  AUTHORS   Gagoski D, Mureev S, Giles N, Johnston W, Dahmer-Heath M, Skalamera 
            D, Gonda TJ, Alexandrov K
  TITLE     Gateway-compatible vectors for high-throughput protein expression in
            pro- and eukaryotic cell-free systems.
  JOURNAL   J Biotechnol. 2015 Feb 10;195:1-7. doi: 
            10.1016/j.jbiotec.2014.12.006. Epub 2014 Dec 19.
  PUBMED    25529348
REFERENCE   2  (bases 1 to 924)
  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: "J 
            Biotechnol."; date: "2015-02-10"; volume: "195:1-7. doi"; pages: " 
            10.1016/j.jbiotec.2014.12.006. Epub 2014 Dec 19"
COMMENT     Sequence Label: pCellFree_G04
FEATURES             Location/Qualifiers
     source          1..924
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             26..49
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      272..318
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(396..412)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(396..412)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(409..431)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    420..436
                     /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(444..450,451..468,469..474))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    489..510
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(627..644)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(878..897)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggacgagcta tacaagacct ctggtcacca ccatcaccac catcaccact aagcggccgc
       61 cctcctcctc ctttcttgtt cctttcacgt cgccttctcg gttgtagctg gcagacgacg
      121 agtcttactt ttacgtgtac ttctctatag atgatgtatg atctctctgc atgcgtgttc
      181 gtgcatgtgt ccgtgtgttg ggtacgcgtg gtaccctgca ggaaggaagc tgagttggct
      241 gctgccaccg ctgagcaata actagtaatt actagcataa ccccttgggg cctctaaacg
      301 ggtcttgagg gggttttttg ctgaaaggag gacagctgat gattgtcatg cttgccatct
      361 gttttcttgc aaggtcagag gaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat
      421 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
      481 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
      541 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
      601 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
      661 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
      721 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
      781 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
      841 cgctcaagtc agaagtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
      901 ggaagctccc tcgtgcgctc tcct
//
