LOCUS       sequence-73161-e         881 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  FX cloning E.coli expression vector with T7 promoter and C-terminal 
            3C protease cleavage site, GFP and 10x His tags.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 881)
  AUTHORS   Geertsma ER, Dutzler R
  TITLE     A versatile and efficient high-throughput cloning tool for 
            structural biology.
  JOURNAL   Biochemistry. 2011 Apr 19;50(15):3272-8. Epub 2011 Mar 25.
  PUBMED    21410291
REFERENCE   2  (bases 1 to 881)
  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: 
            "Biochemistry."; date: "2011-04-19"; volume: "50(15)"; pages: 
            "3272-8. Epub 2011 Mar 25"
COMMENT     Sequence Label: p7XC3GH
FEATURES             Location/Qualifiers
     source          1..881
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(20..37)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(74..103)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             complement(110..820)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGTDELYK"
     primer_bind     complement(150..170)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     765..793
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     CDS             complement(827..850)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
ORIGIN
        1 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gaggatgagt ttttgttcta
       61 gatggtcgac ttaatgatgg tgatgatgat ggtgatgatg atgactagtt ttgtagagct
      121 catccgtgcc atgtgtaatc ccagcagcag ttacaaactc aagaaggacc atgtggtcac
      181 gcttttcgtt gggatctttc gaaagggcag attgtgtcga caggtaatgg ttgtctggta
      241 aaaggacagg gccatcgcca attggagtat tttgttgata atggtctgct agttgaacgg
      301 aaccatcttc aatgttgtgg cgaattttga agttagcttt gattccattc ttttgtttgt
      361 ctgccgtgat gtatacattg tgtgagttat agttgtactc tagtttgtgt ccgagaatgt
      421 ttccatcttc tttaaaatca atacccttta actcgatacg attaacaagg gtatcacctt
      481 caaacttgac ttcagcacgc gtcttgtagt tcccgtcatc tttgaaagat atagtgcgtt
      541 cctgtacata accttcgggc atggcactct tgaaaaagtc atgccgtttc atgtgatccg
      601 gataacggga aaagcattga acaccatagg tcagagtagt gacaagtgtt ggccaaggaa
      661 caggtagttt tccagtagtg caaataaatt taagggtgag ttttccgtat gtagcatcac
      721 cttcaccctc tccactgaca gaaaatttgt gcccattaac atcaccatct aattcaacaa
      781 gaattgggac aactccagtg aaaagttctt ctcctttgct gaattgtgga ccttgaaaca
      841 aaacttctaa tgcagaagag ctgaactagt ggatccccaa a
//
