pKSM713 vector (V005184)

LOCUS       40924_27024        5041 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKSM713, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5041)
  AUTHORS   Maneewannakul S, Maneewannakul K, Ippen-Ihler K.
  TITLE     The pKSM710 vector cassette provides tightly regulated lac and T7lac
            promoters and strategies for manipulating N-terminal protein 
            sequences
  JOURNAL   Plasmid 31 (3), 300-307 (1994)
  PUBMED    8058823
REFERENCE   2  (bases 1 to 5041)
  AUTHORS   Maneewannakul S.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-JAN-1994) Sumit Maneewannakul, Harvard University, 
            Cellular and Developmental Biology, 16 Divinity Avenue, Cambridge, 
            MA 02138, USA
REFERENCE   3  (bases 1 to 5041)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5041)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid"; 
            date: "1994"; volume: "31"; issue: "3"; pages: "300-307"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-JAN-1994) Sumit Maneewannakul, Harvard University, Cellular and 
            Developmental Biology, 16 Divinity Avenue, Cambridge, MA 02138, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5041
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    39..63
                     /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        197..227
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    235..251
                     /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)."
     primer_bind     complement(397..413)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      604..651
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      704..790
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      882..909
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      1009..1057
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     regulatory      1116..1162
                     /label=bacteriophage fd terminator
                     /note="bacteriophage fd terminator"
                     /regulatory_class="terminator"
     terminator      1119..1167
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1295..1750
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      complement(1909..2497)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      complement(2595..2622)
                     /gene="trpA"
                     /regulatory_class="terminator"
     gene            complement(2595..2622)
                     /gene="trpA"
                     /label=trpA
     CDS             complement(2646..3437)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     protein_bind    complement(3829..3850)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3866..4945)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(4946..5023)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."