LOCUS       sequence-86448-e         911 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  E.coli expression of HisYFP tagged human Sp100 fragment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 911)
  AUTHORS   Flotho A, Werner A, Winter T, Frank AS, Ehret H, Melchior F
  TITLE     Recombinant reconstitution of sumoylation reactions in vitro.
  JOURNAL   Methods Mol Biol. 2012;832:93-110. doi: 10.1007/978-1-61779-474-2_5.
  PUBMED    22350878
REFERENCE   2  (bases 1 to 911)
  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: "Methods Mol
            Biol."; date: "2012"; volume: "832:93-110. doi"; pages: " 
            10.1007/978-1-61779-474-2_5"
COMMENT     Sequence Label: pET28a-HisYFP-Sp100
FEATURES             Location/Qualifiers
     source          1..911
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     50..69
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        50..68
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    69..93
                     /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             108..130
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             149..166
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             176..193
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             join(206..208,209..211,212..724)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     complement(251..272)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(512..531)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     859..880
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc
       61 actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt
      121 aagaaggaga tataccatgg gcagcagcca tcatcatcat catcacagca gcggcctggt
      181 gccgcgcggc agccatatgg ctagcatggt gagcaagggc gaggagctgt tcaccggggt
      241 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
      301 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
      361 caagctgccc gtgccctggc ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt
      421 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
      481 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
      541 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
      601 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
      661 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
      721 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
      781 ccccgtgctg ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc
      841 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
      901 cggcatggac g
//
