LOCUS       sequence-382077-        2013 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression plasmid for His tag-based purification of 
            Drosophila melanogaster kinesin heavy chain [1-421] with C-terminal 
            GFP tag and C-terminal 6xHis tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2013)
  AUTHORS   Tas RP, Chazeau A, Cloin BMC, Lambers MLA, Hoogenraad CC, Kapitein 
            LC
  TITLE     Differentiation between Oppositely Oriented Microtubules Controls 
            Polarized Neuronal Transport.
  JOURNAL   Neuron. 2017 Dec 20;96(6):1264-1271.e5. doi: 
            10.1016/j.neuron.2017.11.018. Epub 2017 Nov 30.
  PUBMED    29198755
REFERENCE   2  (bases 1 to 2013)
  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: 
            "10.1016/j.neuron.2017.11.018"; journalName: "Neuron"; date: 
            "2017-12-20- 20"; volume: "96"; issue: "6"; pages: "1264-1271.e5"
COMMENT     Sequence Label: pET28a_DmKHC[1-421]-1xGFP-6xHis
FEATURES             Location/Qualifiers
     source          1..2013
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             join(1270..1272,1273..1275,1276..1986)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1315..1336)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1576..1595)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1923..1944
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1993..2010
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 atgagcgcag aacgagaaat tccagccgag gacagcatca aagtggtctg ccgattccga
       61 ccgctgaacg acagcgaaga gaaggccggc tccaagttcg tggtcaagtt ccccaacaat
      121 gtggaggaga actgcatatc catagcgggc aaggtgtatt tgttcgacaa ggtcttcaaa
      181 ccgaatgcat cccaggaaaa ggtctacaat gaggcggcca agtccattgt tacggatgtc
      241 ctggccgggt acaatggaac gatattcgca tacggtcaga cgtcctccgg aaaaacgcat
      301 acgatggagg gcgtgatcgg ggactccgta aaacagggta tcatacctag gatcgtcaac
      361 gacattttca atcacatcta cgcgatggag gtgaacctgg agtttcacat caaggtctcc
      421 tactacgaga tctacatgga caagattcga gatctgttgg acgtctccaa ggtgaacctt
      481 agtgtgcacg aggataagaa ccgggtgccg tacgtcaagg gcgctacgga acggttcgtc
      541 tcgtcgccgg aggatgtttt cgaggtgatc gaggagggca aatccaatcg tcacatcgct
      601 gtgacaaaca tgaacgagca ttcttcgcga tcccactcag tattccttat caatgtgaag
      661 caggagaacc tggagaacca gaagaaacta tccggcaaac tctacctggt ggatttggcc
      721 ggatccgaga aggtttccaa gactggagcg gagggaaccg ttcttgatga agccaagaac
      781 atcaacaagt cgctgtcggc cttgggcaac gtaatttctg ccctggcgga cggaaacaaa
      841 acgcacatcc cctaccgtga ttccaagcta acgcgcatcc tgcaggagtc gctgggaggc
      901 aacgcacgca caaccatcgt catctgctgc tctccagcca gtttcaacga atctgaaacg
      961 aagtcaacgc tggacttcgg tcgtagagcc aagacagtga agaacgtggt ctgcgttaac
     1021 gaggagctta ctgccgagga atggaagcga cgctatgaaa aggagaagga aaagaacgcc
     1081 cgactaaagg gtaaggtgga gaagcttgag atcgagcttg cgcgctggag agcgggtgaa
     1141 actgttaagg cggaggagca aatcaacatg gaggatctca tggaggcaag cacgcccaac
     1201 ctggaagtgg aggcagcaca gacggcggcg gccgaggccg ctttggccgc ccagcgaacg
     1261 gctgaattca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1321 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1381 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     1441 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     1501 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1561 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     1621 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     1681 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     1741 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     1801 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1861 gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga gaagcgcgat
     1921 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     1981 tacaagctcg agcaccacca ccaccaccac tga
//
