LOCUS       sequence-146508-         922 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses Hook domain of Hook3 in bacteria for purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 922)
  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 922)
  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
FEATURES             Location/Qualifiers
     source          1..922
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(2..79)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     285..304
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     388..407
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        388..406
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    407..431
                     /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             446..468
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             486..503
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             513..530
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             534..566
                     /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(582..605,606..641,642..665)
                     /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(702..730)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
       61 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
      121 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
      181 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
      241 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
      301 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
      361 aggatcgaga tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg
      421 ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata tacatgggca
      481 gcagccatca tcatcatcat cacagcagcg gcctggtgcc gcgcggcagc catatggcta
      541 gcatgactgg tggacagcaa atgggtcgcg gatccgaatt ctggagtcat ccacaatttg
      601 agaagggcgg cggcagtggc ggcggcagtg gcggcagtgc ctggagccac ccgcagttcg
      661 aaaaaggcag tatgagcaaa ggagaagaac ttttcactgg agttgtccca attcttgttg
      721 aattagatgg tgatgttaat gggcacaaat tttctgtccg tggagagggt gaaggtgatg
      781 ctacaaacgg aaaactcacc cttaaattta tttgcactac tggaaaacta cctgttccgt
      841 ggccaacact tgtcactact ctgacctatg gtgttcaatg cttttcccgt tatccggatc
      901 acatgaaacg gcatgacttt tt
//
