Basic Vector Information
- Vector Name:
- pESP-4
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9763 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Grafsky AJ, Lu Q.
- Promoter:
- nmt1
pESP-4 vector Vector Map
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 ℃.
pESP-4 vector Sequence
LOCUS 40924_17621 9763 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pESP-4, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9763)
AUTHORS Grafsky AJ, Lu Q.
TITLE Direct Submission
JOURNAL Submitted (05-FEB-1999) Technical Services, Stratagene, 11011 N.
Torrey Pines Rd., La Jolla, CA 92037, USA
REFERENCE 2 (bases 1 to 9763)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 9763)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
(05-FEB-1999) Technical Services, Stratagene, 11011 N. Torrey Pines
Rd., La Jolla, CA 92037, USA"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9763
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(138..921)
/direction=LEFT
/label=ars1
/note="Schizosaccharomyces pombe autonomously replicating
sequence ars1"
promoter 1296..1400
/label=AmpR promoter
CDS 1401..2258
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 2432..3020
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 3308..3329
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3344..3374
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3382..3398
/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)."
primer_bind 3406..3422
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 3443..3461
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind 3491..3507
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(3915..5006)
/codon_start=1
/label=LEU2
/note="3-isopropylmalate dehydrogenase, required for
leucine biosynthesis"
/translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
DLGGSNSTTEVGDAVAEEVKKILA"
promoter 5962..7119
/label=nmt1 promoter
/note="wild-type nmt1 promoter from Schizosaccharomyces
pombe, conferring strong thiamine-repressible expression"
CDS 7121..7774
/codon_start=1
/label=GST
/note="glutathione S-transferase from Schistosoma
japonicum"
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
CDS 7781..7798
/codon_start=1
/label=thrombin site
/note="thrombin recognition and cleavage site"
/translation="LVPRGS"
CDS 7799..7822
/codon_start=1
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/translation="DYKDDDDK"
promoter complement(8886..8904)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(8914..8930)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 9072..9527
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
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