pAG001 His6-TwinStrep-SUMO-AapCas12b vector (V017800)

LOCUS       .                       9473 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        1..19
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /label="lac operator"
                     /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-β-D-thiogalactopyranoside (IPTG)."
     RBS             59..81
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             100..117
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             127..144
                     /label="thrombin site"
                     /note="thrombin recognition and cleavage site"
     CDS             157..240
                     /label="Twin-Strep-tag"
                     /note="two Strep-Tag®II moieties connected by a linker for
                     enhanced binding to Strep-Tactin®, an engineered form of
                     streptavidin (Schmidt et al., 2013)"
     CDS             241..534
                     /label="SUMO"
                     /note="cleavable ubiquitin-like protein tag"
     terminator      3988..4035
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     promoter        complement(4210..4238)
                     /label="tet promoter"
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     promoter        4351..4455
                     /label="AmpR promoter"
     CDS             4456..5313
                     /label="AmpR"
                     /note="β-lactamase"
     rep_origin      5487..6075
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(6261..6400)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(6505..6693)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(7971..7992)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(8008..9087)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(9088..9165)
                     /label="lacI promoter"