p28BIOH-LIC4 vector (V009769)

LOCUS       40924_2508        7373 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector p28BIOH-LIC4, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7373)
  AUTHORS   Loppnau PA, Graslund S, Arrowsmith CH.
  TITLE     A pipeline to generate recombinant antibodies to human target 
            proteins involved in epigenetic mechanisms
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7373)
  AUTHORS   Loppnau PA, Graslund S, Arrowsmith CH.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-NOV-2012) Structural Genomics Consortium, University 
            of Toronto, MaRS Centre, South tower, 101 College St, Room 700, 
            Toronto, Ontario M5G 1L7, Canada
REFERENCE   3  (bases 1 to 7373)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7373)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-NOV-2012) Structural Genomics Consortium, University of Toronto,
            MaRS Centre, South tower, 101 College St, Room 700, Toronto, Ontario
            M5G 1L7, Canada"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7373
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(563..1375)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      1497..2085
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2271..2413)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2518..2706)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(3481..3502)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3518..4597)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(4598..4675)
                     /label=lacI promoter
     promoter        4984..5002
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5003..5027
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     RBS             5042..5064
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5077..5121
                     /label=AviTag(TM)
                     /note="peptide tag that allows for enzymatic biotinylation"
     misc_feature    5128..5142
                     /note="5'-In-Fusion sequence; add to upstream PCR primer"
     protein_bind    complement(5170..5203)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5218..5663
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             5664..7082
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     misc_feature    7158..7172
                     /note="3'-In-Fusion sequence; add complement to downstream
                     PCR primer"
     CDS             7170..7187
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             7217..7234
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      7301..7348
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"