pSIT vector (V003172)

LOCUS       40924_40492        6101 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Phagemid cloning vector pSIT, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6101)
  AUTHORS   Andreansky M, Hunter E.
  TITLE     Phagemid pSIT permits efficient in vitro mutagenesis and tightly 
            controlled expression in E. coli
  JOURNAL   BioTechniques 16 (4), 626-628 (1994)
  PUBMED    8024782
REFERENCE   2  (bases 1 to 6101)
  AUTHORS   Promega Corporation.
  TITLE     Altered Sites in vitro Mutagenesis System Technical Manual
  JOURNAL   (in) Editor,N.A. (Ed.); TECHNICAL MANUAL, Vol. 1: 1-22; Promega 
            Corporation, Madison, WI (1990)
REFERENCE   3  (bases 1 to 6101)
  AUTHORS   Promega Corporation.
  TITLE     Altered Sites II in vitro Mutagenesis System Technical Manual
  JOURNAL   (in) Editor,N.A. (Ed.); TECHNICAL MANUAL, Vol. 2: 1-33; Promega 
            Corporation, Madison, WI (1994)
REFERENCE   4  (bases 1 to 6101)
  AUTHORS   Studier FW, Rosenberg AH, Dunn JJ, Dubendorff JW.
  TITLE     Use of T7 RNA polymerase to direct expression of cloned genes
  JOURNAL   Meth. Enzymol. 185, 60-89 (1990)
  PUBMED    2199796
REFERENCE   5  (bases 1 to 6101)
  AUTHORS   Hunter E.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-NOV-1994) Eric Hunter, Microbiology, University of 
            Alabama at Birmingham, 845 19th St.S., Bevill Biomedical Research 
            Building, Rm 256, Birmingham, AL 35294, USA
REFERENCE   6  (bases 1 to 6101)
  TITLE     Direct Submission
REFERENCE   7  (bases 1 to 6101)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "BioTechniques"; date: "1994"; volume: "16"; issue: "4"; pages: 
            "626-628"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "(in) 
            Editor,N.A. (Ed.)"; volume: " TECHNICAL MANUAL, Vol. 1"; pages: " 
            1-22; Promega Corporation, Madison, WI (1990"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "(in) 
            Editor,N.A. (Ed.)"; volume: " TECHNICAL MANUAL, Vol. 2"; pages: " 
            1-33; Promega Corporation, Madison, WI (1994"
COMMENT     SGRef: number: 4; type: "Journal Article"; journalName: "Meth. 
            Enzymol. 185, 60-89 (1990)"
COMMENT     SGRef: number: 5; type: "Journal Article"; journalName: "Submitted 
            (14-NOV-1994) Eric Hunter, Microbiology, University of Alabama at 
            Birmingham, 845 19th St.S., Bevill Biomedical Research Building, Rm 
            256, Birmingham, AL 35294, USA"
COMMENT     SGRef: number: 6; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6101
                     /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)."
     RBS             78..100
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             107..139
                     /label=T7 tag (gene 10 leader)
                     /note="leader peptide from bacteriophage T7 gene 10"
     terminator      205..252
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        473..501
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             549..1736
                     /label=TcR
                     /note="tetracycline efflux protein"
     primer_bind     1957..1973
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1989..2005)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2013..2029)
                     /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(2037..2067)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2082..2103)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(2119..3198)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(3199..3276)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     rep_origin      3518
                     /note="ColE1 based origin of DNA replication, that proceeds
                     counterclockwise from T (nucleotide 3518); ROP that down 
                     regulates plasmid copy number has been deleted in pSIT, 
                     providing higher DNA yields"
     rep_origin      complement(3520..4108)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4577..4951)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene;
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     promoter        complement(5136..5240)
                     /label=AmpR promoter
     rep_origin      complement(5576..6031)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"