GST-SBP vector (V009929)

Basic Vector Information

      • Vector Name:
      • GST-SBP
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 4779 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Jerlstrom-Hultqvist J, Stadelmann B, Birkestedt S, Hellman U, Svard SG.

GST-SBP vector Vector Map

GST-SBP4779 bp600120018002400300036004200AmpR promoterAmpRoriCAP binding sitelac promoterlac operatorM13 revT3 promoter3'UTRPreScission protease cleavage siteglutathione S-transferaseSBP-tag3'UTRT7 promoterM13 fwdf1 ori

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

GST-SBP vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                4779 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector GST-SBP, complete sequence.
ACCESSION   JQ772529
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4779)
  AUTHORS   Jerlstrom-Hultqvist J, Stadelmann B, Birkestedt S, Hellman U, Svard 
            SG.
  TITLE     Plasmid vectors for proteomic analyses in giardia: purification of 
            virulence factors and analysis of the proteasome
  JOURNAL   Eukaryotic Cell 11 (7), 864-873 (2012)
  PUBMED    22611020
REFERENCE   2  (bases 1 to 4779)
  AUTHORS   Jerlstrom-Hultqvist J, Stadelmann B, Birkestedt S, Hellman U, Svard 
            SG.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-MAR-2012) Department of Cell and Molecular Biology, 
            Uppsala University, BMC, Box 596, Uppsala 751 24, Sweden
REFERENCE   3  (bases 1 to 4779)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4779
                     /organism="Cloning vector GST-SBP"
                     /mol_type="other DNA"
                     /db_xref="taxon:1195238"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             129..989
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             327..989
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="AFK93521.1"
                     /translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHL
                     TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA
                     IPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI
                     ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2072..2102
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2171..2189
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     3'UTR           2208..3227
                     /note="glutamate dehydrogenase 3' UTR; derived from Giardia
                     intestinalis"
     CDS             complement(2325..2933)
                     /codon_start=1
                     /product="puromycin N-acetyltransferase"
                     /label=puromycin N-acetyltransferase
                     /protein_id="AFK93522.1"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVECPKDRATWCMTRKPGALE"
     CDS             complement(2334..2933)
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     5'UTR           2934..2981
                     /note="glutamate dehydrogenase 5' UTR; derived from Giardia
                     intestinalis"
     5'UTR           2988..3187
                     /note="ornithine carbamoyltransferase 5' UTR; derived from 
                     Giardia intestinalis"
     misc_feature    3239..3262
                     /label=PreScission protease cleavage site
                     /note="PreScission protease cleavage site"
     CDS             3239..3262
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     misc_feature    3263..3919
                     /label=glutathione S-transferase
                     /note="glutathione S-transferase"
     CDS             3263..3913
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="SPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKF
                     ELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYG
                     VSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLY
                     MDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     misc_feature    3926..4039
                     /label=SBP-tag
                     /note="SBP-tag"
     CDS             3926..4039
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     3'UTR           4046..4121
                     /note="alpha-1 giardin 3' UTR; derived from Giardia 
                     intestinalis"
     promoter        complement(4136..4154)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4164..4180)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4322..4777
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"

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