pCW-LIC vector (V008105)

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LOCUS       40924_13880        6980 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pCW-LIC, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6980)
  AUTHORS   Loppnau P, Kozieradzki I, Arrowsmith C.
  TITLE     Expression vectors for high-throughput In-Fusion cloning
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6980)
  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 6980)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6980)
  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            "Unpublished"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..6980
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..465
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        492..596
                     /label=AmpR promoter
     CDS             597..1454
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1628..2216
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2402..2544)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2953..3126)
                     /label=lacZ-alpha
                     /note="LacZ-alpha fragment of beta-galactosidase"
     protein_bind    complement(3146..3162)
                     /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)."
     promoter        complement(3170..3200)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3215..3236)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3252..4331)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(4332..4409)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     protein_bind    4461..4482
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4497..4527
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    4535..4551
                     /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)."
     promoter        4606..4634
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    4640..4660
                     /label=lac repressor protein binding site
                     /bound_moiety="lac repressor protein"
     protein_bind    4642..4658
                     /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)."
     regulatory      4701..4729
                     /label=Ptac
                     /note="Ptac"
                     /regulatory_class="promoter"
     promoter        4701..4729
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     regulatory      4701..4706
                     /label=derived from trp promoter
                     /note="derived from trp promoter"
                     /regulatory_class="minus_35_signal"
     regulatory      4723..4729
                     /label=derived from lacUV5 promoter
                     /note="derived from lacUV5 promoter"
                     /regulatory_class="minus_10_signal"
     protein_bind    4735..4755
                     /label=lac repressor protein binding site
                     /bound_moiety="lac repressor protein"
     protein_bind    4737..4753
                     /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)."
     misc_feature    4784..4798
                     /note="5'-cloning sequence; add to upstream PCR primer"
     protein_bind    complement(4826..4859)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4874..5319
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             5320..6738
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     misc_feature    6814..6828
                     /note="3'-cloning sequence; add complement to downstream
                     PCR primer"
     regulatory      6862..6889
                     /label=trp operon transcription termination sequence
                     /note="trp operon transcription termination sequence"
                     /regulatory_class="terminator"