LOCUS       sequence-146511-         636 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses artificially dimerized Hook domain of Hook3 in bacteria 
            for purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 636)
  AUTHORS   Schroeder CM, Vale RD
  TITLE     Assembly and activation of dynein-dynactin by the cargo adaptor 
            protein Hook3.
  JOURNAL   J Cell Biol. 2016 Aug 1;214(3):309-18. doi: 10.1083/jcb.201604002.
  PUBMED    27482052
REFERENCE   2  (bases 1 to 636)
  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"; doi: "10.1083/jcb"; 
            journalName: "J Cell Biol"; date: "2016-08-1- 1"; volume: "214"; 
            issue: "3"; pages: "309-18"
COMMENT     Sequence Label: pET28a-Hook3 aa 1-160-GCN4
FEATURES             Location/Qualifiers
     source          1..636
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    72..96
                     /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             111..133
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             151..168
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             178..195
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             199..231
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(247..270,271..306,307..330)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="WSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     primer_bind     complement(367..395)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa nttaatacna
       61 ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac
      121 tttaagaagg agatatacat gggcagcagc catcatcatc atcatcacag cagcggcctg
      181 gtgccgcgcg gcagccatat ggctagcatg actggtggac agcaaatggg tcgcggatcc
      241 gaattctgga gtcatccaca atttgagaag ggcggcggca gtggcggcgg cagtggcggc
      301 agtgcctgga gccacccgca gttcgaaaaa ggcagtatga gcaaaggaga agaacttttc
      361 actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct
      421 gtccgtggag agggtgaagg tgatgctaca aacggaaaac tcacccttaa atttatttgc
      481 actactggaa aactacctgt tccgtggcca acacttgtca ctactctgac ctatggtgtt
      541 caatgctttt cccgttatcc ggatcacatg aaacggcatg actttttcaa gagtgccatg
      601 cccgaaggtt atgtacagga acgcactata tctttc
//
