LOCUS       sequence-214272-         934 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses MBP-GFP-DHFR; construct generated for pull-down and 
            localization experiments.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 934)
  AUTHORS   Caldwell RM, Bermudez JG, Thai D, Aonbangkhen C, Schuster BS, 
            Courtney T, Deiters A, Hammer DA, Chenoweth DM, Good MC
  TITLE     Optochemical Control of Protein Localization and Activity within 
            Cell-like Compartments.
  JOURNAL   Biochemistry. 2018 May 8;57(18):2590-2596. doi: 
            10.1021/acs.biochem.8b00131. Epub 2018 Apr 19.
  PUBMED    29671583
REFERENCE   2  (bases 1 to 934)
  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: 
            "10.1021/acs.biochem.8b00131"; journalName: "Biochemistry"; date: 
            "2018-05-8- 8"; volume: "57"; issue: "18"; pages: "2590-2596"
COMMENT     Sequence Label: pJB03
FEATURES             Location/Qualifiers
     source          1..934
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             49..60
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             103..123
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(148..150,151..153,154..864)
                     /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(193..214)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(454..473)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     801..822
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 aattcgagct cgaacaacaa caacaataac aataacaaca acctcgggat cgagggaagg
       61 atttcaacta gtggatctgg tggtggtggc ggttccatga gcgagaatct ttattttcag
      121 ggatccagcg gcggttctag cggccgcatg gtgagcaagg gcgaggagct gttcaccggg
      181 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
      241 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
      301 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
      361 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
      421 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
      481 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
      541 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
      601 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
      661 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
      721 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac
      781 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
      841 ctcggcatgg acgagctgta caagctcgag gacgagctgt ataaggccgg cctggagtct
      901 agaatcagtc tgattgcggc gttagcggta gatc
//
