pTRC-LIC vector (V002552)

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LOCUS       40924_43888        6148 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTRC-LIC, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6148)
  AUTHORS   Loppnau P, Kozieradzki I, Arrowsmith C.
  TITLE     Expression vectors for high-throughput In-Fusion cloning
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6148)
  AUTHORS   Loppnau P, Kozieradzki I, Arrowsmith C.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-FEB-2007) Structural Genomics Consortium, University 
            of Toronto, 100 College Street, Toronto, ON M5G1L5, Canada
REFERENCE   3  (bases 1 to 6148)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6148)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (27-FEB-2007) Structural Genomics Consortium, University of Toronto,
            100 College Street, Toronto, ON M5G1L5, Canada"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6148
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..92
                     /label=AmpR promoter
     CDS             93..950
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1124..1712
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(1898..2038)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        2224..2301
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             2302..3381
                     /label=lacI
                     /note="lac repressor"
     promoter        3616..3645
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    3653..3669
                     /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)."
     misc_feature    3678..3692
                     /note="5'-LIC sequence; add to upstream PCR primer"
     protein_bind    complement(3720..3753)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3768..4213
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             4214..5632
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     misc_feature    5708..5722
                     /note="3'-LIC sequence; add complement to downstream PCR
                     primer"
     rRNA            5804..5923
                     /gene="rrnB"
                     /product="5S ribosomal RNA"
                     /label=rrnB
     terminator      5924..6010
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      6102..6129
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"