LOCUS       sequence-73082-e         944 bp    DNA     linear   UNA 14-APR-2025
DEFINITION  FX cloning E.coli expression vector with T7 promoter and N-terminal 
            10x His tag and 3C protease cleavage site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 944)
  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 944)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: p7XNH3
FEATURES             Location/Qualifiers
     source          1..944
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             6..28
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             39..68
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             69..92
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(141..446)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(243..262)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
ORIGIN
        1 aatattttgt ttaactttaa gaaggagata tacatatgca ccatcatcat caccatcatc
       61 atcatcattt agaagttttg tttcaaggtc caagtagaag agcgagctgc agactggctg
      121 tgtataaggg agcctgacat ttatattccc cagaacatca ggttaatggc gtttttgatg
      181 tcattttcgc ggtggctgag atcagccact tcttccccga taacggagac cggcacactg
      241 gccatatcgg tggtcatcat gcgccagctt tcatccccga tatgcaccac cgggtaaagt
      301 tcacgggaga ctttatctga cagcagacgt gcactggcca gggggatcac catccgtcgc
      361 ccgggcgtgt caataatatc actctgtaca tccacaaaca gacgataacg gctctctctt
      421 ttataggtgt aaaccttaaa ctgcatttca ccagcccctg ttctcgtcag caaaagagcc
      481 gttcatttca ataaaccggg cgacctcagc catcccttcc tgattttccg ctttccagcg
      541 ttcggcacgc agacgacggg cttcattctg catggttgtg cttaccagac cggagatatt
      601 gacatcatat atgccttgag caactgatag ctgtcgctgt caactgtcac tgtaatacgc
      661 tgcttcatag catacctctt tttgacatac ttcgggtata catatcagta tatattctta
      721 taccgcaaaa atcagcgcgc aaatacgcat actgttatct ggcttttagt aagccggatc
      781 cacgcgttta cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc
      841 tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca
      901 ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggg
//
