LOCUS       sequence-93632-e         808 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  expresses CKK domain of CAMSAP3 in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 808)
  AUTHORS   Hendershott MC, Vale RD
  TITLE     Regulation of microtubule minus-end dynamics by CAMSAPs and 
            Patronin.
  JOURNAL   Proc Natl Acad Sci U S A. 2014 Apr 22;111(16):5860-5. doi: 
            10.1073/pnas.1404133111. Epub 2014 Mar 26.
  PUBMED    24706919
REFERENCE   2  (bases 1 to 808)
  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: 
            "10.1073/pnas.1404133111"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2014-04-22- 22"; volume: "111"; issue: "16"; pages: "5860-5"
COMMENT     Sequence Label: pET-28a+GFP-CAMSAP3 CKK
FEATURES             Location/Qualifiers
     source          1..808
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(join(16..39,40..75,76..99))
                     /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"
     CDS             complement(115..147)
                     /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             complement(151..168)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(178..195)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             213..235
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    250..274
                     /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)."
     primer_bind     complement(274..293)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(275..293)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(377..396)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        602..679
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(699..718)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tttgctcata ctgccttttt cgaactgcgg gtggctccag gcactgccgc cactgccgcc
       61 gccactgccg ccgcccttct caaattgtgg atgactccag aattcggatc cgcgacccat
      121 ttgctgtcca ccagtcatgc tagccatatg gctgccgcgc ggcaccaggc cgctgctgtg
      181 atgatgatga tgatggctgc tgcccatgta tatctccttc ttaaagttaa acaaaattat
      241 ttctagaggg gaattgttat ccgctcacaa ttcccctata gtgagtcgta ttaatttcgc
      301 gggatcgaga tctcgatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca
      361 ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac
      421 ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga
      481 ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc
      541 aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg tcgagatccc
      601 ggacaccatc gaatggcgca aaacctttcg cggtatggca tgatagcgcc cggaagagag
      661 tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg
      721 tgtctcttat cagaccgttt cccgcgtggn gaaccaggcc agccacgttt ctgcgaaaac
      781 gcggggaaaa gtggaaagcg gcgatggc
//
