LOCUS       sequence-59812-e         822 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  pKW32 converted Gateway Destination vector for expressing hemoglobin
            tagged fusion proteins in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 822)
  AUTHORS   Guo F, Chiang MY, Wang Y, Zhang YZ
  TITLE     An in vitro recombination method to convert restriction- and 
            ligation-independent expression vectors.
  JOURNAL   Biotechnol J. 2008 Mar . 3(3):370-7.
  PUBMED    18064608
REFERENCE   2  (bases 1 to 822)
  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 
            J. 2008 Mar . 3(3):370-7."
COMMENT     Sequence Label: pEZYHb
FEATURES             Location/Qualifiers
     source          1..822
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     83..101
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(125..139,140..145,146..162,163..169)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    145..161
                     /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)."
     protein_bind    177..193
                     /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)."
     RBS             212..223
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             242..259
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             731..748
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
ORIGIN
        1 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
       61 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcacctcga
      121 gaaatcataa aaaatttatt tgctttgtga gcggataaca attataatag attcaattgt
      181 gagcggataa caatttcaca cagaattcat taaagaggag aaattaacta tgagaggatc
      241 gcatcaccat caccatcacg gatccatgtt agaccagcaa accattaaca tcatcaaagc
      301 cactgttcct gtattgaagg agcatggcgt taccattacc acgacttttt ataaaaactt
      361 gtttgccaaa caccctgaag tacgtccttt gtttgatatg ggtcgccaag aatctttgga
      421 gcagcctaag gctttggcga tgacggtatt ggcggcagcg caaaacattg aaaatttgcc
      481 agctattttg cctgcggtca aaaaaattgc agtcaaacat tgtcaagcag gcgtggcagc
      541 agcgcattat ccgattgtcg gtcaagaatt gttgggtgcg attaaagaag tattgggcga
      601 tgccgcaacc gatgacattt tggacgcgtg gggcaaggct tatggcgtga ttgcagatgt
      661 gtttattcaa gtggaagcag atttgtacgc tcaagcggtt gaaccaggaa tatcaggcgg
      721 aggcggaggc ttggtccctc gagggagtga acataataca agtttgtaca aaaaaagctg
      781 aacgagaaac gtaaaatgat ataaatatca atatattaaa tt
//
