LOCUS       sequence-257467-        1048 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression of His6-GFP-C-LACE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1048)
  AUTHORS   Hofmann R, Akimoto G, Wucherpfennig TG, Zeymer C, Bode JW
  TITLE     Lysine acylation using conjugating enzymes for site-specific 
            modification and ubiquitination of recombinant proteins.
  JOURNAL   Nat Chem. 2020 Nov;12(11):1008-1015. doi: 10.1038/s41557-020-0528-y.
            Epub 2020 Sep 14.
  PUBMED    32929246
REFERENCE   2  (bases 1 to 1048)
  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: 
            "10.1038/s41557-020-0528-y"; journalName: "Nat Chem"; date: 
            "2020-11"; volume: "12"; issue: "11"; pages: "1008-1015"
COMMENT     Sequence Label: pET28-His6-GFP-C-LACE
FEATURES             Location/Qualifiers
     source          1..1048
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             5..27
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             46..63
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             76..96
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             97..810
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             862..879
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(932..950)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      946..993
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 taattttgtt taactttaag aaggagatat accatgggca gttcacatca tcatcatcat
       61 cacagcagcg gtgccgaaaa cttatatttt cagggtatgc gtaaaggcga agaattattt
      121 acgggcgtag tgcctatttt agtggaactg gatggcgatg tgaatggaca caagtttagc
      181 gtgcgtggcg aaggtgaagg cgatgcgaca aatggcaagc tgacgctgaa atttatctgt
      241 acgaccggta aactgccggt tccctggccg accctggtta ccaccctgac atacggtgtg
      301 caatgcttcg cccgctatcc agatcacatg aaacagcatg actttttcaa gagcgcgatg
      361 cccgaaggct acgttcaaga gcgtaccatc agcttcaaag atgacggcac ttataaaacc
      421 cgcgcggaag taaagtttga aggtgataca ctggtcaatc gcatcgaact gaagggtatt
      481 gactttaaag aagatggcaa catcttaggc cataaattag agtataactt caactcacac
      541 aatgtgtaca ttacggcaga caaacagaag aacggtatta aggccaactt caagattcgt
      601 cataacgtag aagatggctc ggtccagtta gcggatcact atcaacagaa tacccctatt
      661 ggcgatggtc cagttctgct gccggataat cactacctga gtacgcagtc agtcctgtcg
      721 aaagacccga atgagaaacg cgaccatatg gttctgttag aattcgtaac tgccgcaggc
      781 attacccacg gcatggacga gctgtacaaa ggtagcggtc cgcgtaaggt tatcaaaatg
      841 gaatcggaag agtaactcga gcaccaccac caccaccact gagatccggc tgctaacaaa
      901 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
      961 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggattg
     1021 gcgaatggna cgcgccctgt agcggcgc
//
