pSET6s vector (V003381)

Basic Vector Information

pSET4s, pSET5s, and pSET6s are three thermosensitive (Ts) suicide vectors used for gene replacement in Streptococcus suis. These vectors could be propagated at 37°C in Escherichia coli, but their replication was blocked above 37°C in S. suis. orfC, replication regulatory protein; repATs, thermosensitive replication initiation–termination protein.

Vector Name:
pSET6s
Antibiotic Resistance:
Chloramphenicol
Length:
4438 bp
Type:
Thermosensitive suicide vector
Replication origin:
ori
Source/Author:
Takamatsu D, Osaki M, Sekizaki T.
Growth Strain(s):
DH5alpha
Growth Temperature:
37℃

pSET6s vector Map

pSET6s4438 bp600120018002400300036004200Factor Xa siteOrfCRepAtsM13 fwdMCSM13 revlac operatorlac promoterCAP binding siteoriChloramphenicol acetyltransferase

pSET6s vector Sequence

LOCUS       V003381                 4438 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003381
VERSION     V003381
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4438)
  AUTHORS   Takamatsu D, Osaki M, Sekizaki T.
  TITLE     Thermosensitive suicide vectors for gene replacement in
            Streptococcus suis
  JOURNAL   Plasmid 46 (2), 140-148 (2001)
   PUBMED   11591139
REFERENCE   2  (bases 1 to 4438)
  AUTHORS   Takamatsu D, Osaki M, Sekizaki T.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-FEB-2001) Daisuke Takamatsu, National Institute of
            Animal Health, Laboratory of Molecular Bacteriology; Kannondai
            3-1-1, Tsukuba, Ibaraki 305-0856, Japan
            (E-mail:p1013dt@niah.affrc.go.jp, Tel:81-298-38-7743,
            Fax:81-298-38-7743)
REFERENCE   3  (bases 1 to 4438)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4438)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid";
            date: "2001"; volume: "46"; issue: "2"; pages: "140-148"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (08-FEB-2001) Daisuke Takamatsu, National Institute of Animal
            Health, Laboratory of Molecular Bacteriology"; volume: " Kannondai
            3-1-1, Tsukuba, Ibaraki 305-0856, Japan
            (E-mail:p1013dt@niah.affrc.go.jp, Tel:81-298-38-7743, Fax"; pages:
            "81-298-38-7743"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(133..144)
                     /label="Factor Xa site"
                     /note="Factor Xa recognition and cleavage site"
     CDS             528..689
                     /codon_start=1
                     /gene="orfC"
                     /product="OrfC"
                     /label="orfC"
                     /protein_id="BAB64888.1"
                     /translation="MVISESKKRVMISLTKEQDKKLTDMAKQKGFSKSAVAALAIEEYA
                     RKESEQKK"
     gene            528..689
                     /gene="orfC"
                     /label="orfC"
     CDS             756..1454
                     /codon_start=1
                     /gene="repAts"
                     /product="RepAts"
                     /label="repAts"
                     /protein_id="BAB64889.1"
                     /translation="MAIKNTKARNFGFLLYPDSIPNDWKEKLESLGVSMAVSPLHDMDE
                     KKDKDTWNNSNIIQNGKHYKKPHYHVIYIARNPVTIESVRNKIKRKLGNSSVAHVEILD
                     YIKGSYEYLTHESKDAIAKNKHIYDKKDILNINDFDIDRYITLDESQKRELKNLLLDIV
                     DDYNLVNTKDLMAFIRLRGAEFGILNTNDVKDIVSTNSSAFRLWFEGNYQCGYRASYAK
                     VLDAETGEIK"
     gene            756..1454
                     /gene="repAts"
                     /label="repAts"
     primer_bind     2036..2052
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    2053..2109
                     /label="MCS"
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2122..2138)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(2146..2162)
                     /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(2170..2200)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2215..2236)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2524..3112)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(3575..4222)
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"

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