LOCUS       sequence-342251-        2730 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  LgBit fused to LAF-1 RGG domain with N-terminal MBP tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2730)
  AUTHORS   Guan M, Garabedian MV, Leutenegger M, Schuster BS, Good MC, Hammer 
            DA
  TITLE     Incorporation and Assembly of a Light-Emitting Enzymatic Reaction 
            into Model Protein Condensates.
  JOURNAL   Biochemistry. 2021 Oct 14. doi: 10.1021/acs.biochem.1c00373.
  PUBMED    34648259
REFERENCE   2  (bases 1 to 2730)
  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: 
            "Biochemistry. 2021 Oct 14. doi: 10.1021/acs.biochem.1c00373."
COMMENT     Sequence Label: MBP-RGG-LgBit-RGG
FEATURES             Location/Qualifiers
     source          1..2730
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             4..21
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      18..27
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             34..1134
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1108..1131
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1165..1188
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2182..2199
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2710..2727
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 atgcaccatc accatcacca tggcggcgga tctatgaaaa tcgaagaagg taaactggta
       61 atctggatta acggcgataa aggctataac ggtctcgctg aagtcggtaa gaaattcgag
      121 aaagataccg gaattaaagt caccgttgag catccggata aactggaaga gaaattccca
      181 caggttgcgg caactggcga tggccctgac attatcttct gggcacacga ccgctttggt
      241 ggctacgctc aatctggcct gttggctgaa atcaccccgg acaaagcgtt ccaggacaag
      301 ctgtatccgt ttacctggga tgccgtacgt tacaacggca agctgattgc ttacccgatc
      361 gctgttgaag cgttatcgct gatttataac aaagatctgc tgccgaaccc gccaaaaacc
      421 tgggaagaga tcccggcgct ggataaagaa ctgaaagcga aaggtaagag cgcgctgatg
      481 ttcaacctgc aagaaccgta cttcacctgg ccgctgattg ctgctgacgg gggttatgcg
      541 ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa cgctggcgcg
      601 aaagcgggtc tgaccttcct ggttgacctg attaaaaaca aacacatgaa tgcagacacc
      661 gattactcca tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac catcaacggc
      721 ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt atggtgtaac ggtactgccg
      781 accttcaagg gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg tattaacgcc
      841 gccagtccga acaaagagct ggcaaaagag ttcctcgaaa actatctgct gactgatgaa
      901 ggtctggaag cggttaataa agacaaaccg ctgggtgccg tagcgctgaa gtcttacgag
      961 gaagagttgg tgaaagatcc gcgtattgcc gccactatgg aaaacgccca gaaaggtgaa
     1021 atcatgccga acatcccgca gatgtccgct ttctggtatg ccgtgcgtac tgcggtgatc
     1081 aacgccgcca gcggtcgtca gactgtcgat gaagccctga aagacgcgca gaccggtggg
     1141 ggagggagtg gcggtggtgg cagccttgaa gtcctctttc aggggccggg tatggaatcc
     1201 aatcaatcca ataacggtgg ctcgggtaat gccgctctga atcgtggtgg ccgctacgtg
     1261 ccgccgcatc tgcgtggtgg cgatggtggt gcagcagcag ctgcatctgc tggcggtgat
     1321 gaccgtcgcg gcggtgcggg cggtggcggt tatcgtcgcg gcggtggcaa tagtggtggc
     1381 ggtggcggtg gcggttatga tcgtggttac aatgacaacc gtgatgaccg cgataaccgt
     1441 ggcggttccg gcggttacgg tcgtgatcgt aattatgaag accgtggtta caacggcggt
     1501 ggcggtggcg gtggcaatcg tggctataac aataaccgtg gtggcggtgg cggtggctac
     1561 aatcgccagg atcgtggtga cggtggcagc tctaactttt cgcgcggtgg ctataataac
     1621 cgtgatgaag gcagcgacaa ccgcggtagt ggccgttcct acaataacga tcgtcgcgac
     1681 aatggtggcg atggcgaatt cgtttttacg cttgaagatt tcgttggtga ttgggagcaa
     1741 actgcggcat ataatttgga ccaagttttg gaacaaggtg gtgtaagtag tcttcttcag
     1801 aacttagctg tctccgtgac acccattcag cgcatcgtta gatcgggtga aaacgccctg
     1861 aagatagata ttcacgttat tattccctat gagggcttaa gtgcagatca aatggcccaa
     1921 atcgaggagg ttttcaaggt cgtgtatcca gtggacgatc accatttcaa agtcatttta
     1981 ccgtatggga ctttagtaat tgacggagta actcccaata tgttgaacta ttttggaaga
     2041 ccatacgagg gcatagcggt tttcgatggt aaaaagatta cggtgacggg tacgctttgg
     2101 aatggtaaca aaattataga cgagcggtta ataaccccgg atgggtcaat gttattcaga
     2161 gtcactatta acagtgaatt cttagtacct cgcgggtcaa tggaatccaa tcaatccaat
     2221 aacggtggct cgggtaatgc cgctctgaat cgtggtggcc gctacgtgcc gccgcatctg
     2281 cgtggtggcg atggtggtgc agcagcagct gcatctgctg gcggtgatga ccgtcgcggc
     2341 ggtgcgggcg gtggcggtta tcgtcgcggc ggtggcaata gtggtggcgg tggcggtggc
     2401 ggttatgatc gtggttacaa tgacaaccgt gatgaccgcg ataaccgtgg cggttccggc
     2461 ggttacggtc gtgatcgtaa ttatgaagac cgtggttaca acggcggtgg cggtggcggt
     2521 ggcaatcgtg gctataacaa taaccgtggt ggcggtggcg gtggctacaa tcgccaggat
     2581 cgtggtgacg gtggcagctc taacttttcg cgcggtggct ataataaccg tgatgaaggc
     2641 agcgacaacc gcggtagtgg ccgttcctac aataacgatc gtcgcgacaa tggtggcgat
     2701 ggcctcgagc accaccacca ccaccactga
//
