LOCUS       sequence-146515-         949 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses a triple mutant truncation of Hook3 in bacteria for 
            purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 949)
  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 949)
  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 N68A, W69A, K77A
FEATURES             Location/Qualifiers
     source          1..949
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     63..82
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        63..81
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    82..106
                     /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             121..143
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             161..178
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             188..205
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             209..241
                     /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(257..280,281..316,317..340)
                     /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(377..405)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa
       61 attaatacga ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat
      121 tttgtttaac tttaagaagg agatatacat gggcagcagc catcatcatc atcatcacag
      181 cagcggcctg gtgccgcgcg gcagccatat ggctagcatg actggtggac agcaaatggg
      241 tcgcggatcc gaattctgga gtcatccaca atttgagaag ggcggcggca gtggcggcgg
      301 cagtggcggc agtgcctgga gccacccgca gttcgaaaaa ggcagtatga gcaaaggaga
      361 agaacttttc actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca
      421 caaattttct gtccgtggag agggtgaagg tgatgctaca aacggaaaac tcacccttaa
      481 atttatttgc actactggaa aactacctgt tccgtggcca acacttgtca ctactctgac
      541 ctatggtgtt caatgctttt cccgttatcc ggatcacatg aaacggcatg actttttcaa
      601 gagtgccatg cccgaaggtt atgtacagga acgcactata tctttcaaag atgacgggac
      661 ctacaagacg cgtgctgaag tcaagtttga aggtgatacc cttgttaatc gtatcgagtt
      721 aaagggtatt gattttaaag aagatggaaa cattcttgga cacaaactcg aatacaactt
      781 taactcacac aatgtataca tcacggcaga caaacaaaag aatggaatca aagctaactt
      841 caaaattcgc cacaacgttg aagatggttc cgttcaacta gcagaccatt atcaacaaaa
      901 tactccaatt ggcgatggcc ctgtcctttt accagacaac cattacctg
//
