LOCUS       sequence-146526-         946 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  used for expression in E. coli and purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 946)
  AUTHORS   Schroeder CM, Ostrem JM, Hertz NT, Vale RD
  TITLE     A Ras-like domain in the light intermediate chain bridges the dynein
            motor to a cargo-binding region.
  JOURNAL   Elife. 2014 Oct 1;3:e03351. doi: 10.7554/eLife.03351.
  PUBMED    25272277
REFERENCE   2  (bases 1 to 946)
  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: "Elife."; 
            date: "2014-10-1"; volume: "3:e03351. doi"; pages: " 
            10.7554/eLife.03351"
COMMENT     Sequence Label: pET28a-RILP
FEATURES             Location/Qualifiers
     source          1..946
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     69..88
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        69..87
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    88..112
                     /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             127..149
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             168..185
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             195..212
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             join(219..242,243..278,279..302)
                     /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(339..367)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc
       61 cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga
      121 aataattttg tttaacttta agaaggagat ataccatggg cagcagccat catcatcatc
      181 atcacagcag cggcctggtg ccgcgcggca gccatatgtg gagtcatcca caatttgaga
      241 agggcggcgg cagtggcggc ggcagtggcg gcagtgcctg gagccacccg cagttcgaaa
      301 aaggcagtat gagcaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat
      361 tagatggtga tgttaatggg cacaaatttt ctgtccgtgg agagggtgaa ggtgatgcta
      421 caaacggaaa actcaccctt aaatttattt gcactactgg aaaactacct gttccgtggc
      481 caacacttgt cactactctg acctatggtg ttcaatgctt ttcccgttat ccggatcaca
      541 tgaaacggca tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaacgcacta
      601 tatctttcaa agatgacggg acctacaaga cgcgtgctga agtcaagttt gaaggtgata
      661 cccttgttaa tcgtatcgag ttaaagggta ttgattttaa agaagatgga aacattcttg
      721 gacacaaact cgaatacaac tttaactcac acaatgtata catcacggca gacaaacaaa
      781 agaatggaat caaagctaac ttcaaaattc gccacaacgt tgaagatggt tccgttcaac
      841 tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca
      901 accattacct gtcgacacaa tctgtccttt cgaaagatcc caacga
//
