Basic Vector Information
- Vector Name:
- pMBP-parallel1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6724 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Sheffield P, Garrard S, Derewenda Z.
pMBP-parallel1 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 ℃.
pMBP-parallel1 vector Sequence
LOCUS 40924_29906 6724 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pMBP-parallel1, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6724)
AUTHORS Sheffield P, Garrard S, Derewenda Z.
TITLE Overcoming expression and purification problems of RhoGDI using a
family of 'parallel' expression vectors
JOURNAL Protein Expr. Purif. 15 (1), 34-39 (1999)
PUBMED 10024467
REFERENCE 2 (bases 1 to 6724)
AUTHORS Sheffield PJ, Garrard SM, Derewenda ZS.
TITLE Direct Submission
JOURNAL Submitted (05-OCT-1998) Molecular Physiology and Biological Physics,
University of Virginia, 4215 Jordan Hall, 1300 Jefferson Park
Avenue, Charlottesville, Virginia 22908, USA
REFERENCE 3 (bases 1 to 6724)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6724)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Protein
Expr. Purif."; date: "1999"; volume: "15"; issue: "1"; pages:
"34-39"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(05-OCT-1998) Molecular Physiology and Biological Physics,
University of Virginia, 4215 Jordan Hall, 1300 Jefferson Park
Avenue, Charlottesville, Virginia 22908, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6724
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 3..80
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
CDS 81..1160
/codon_start=1
/label=lacI
/note="lac repressor"
/translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
protein_bind 1176..1197
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 1406..1434
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 1442..1458
/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)."
CDS 1528..2628
/codon_start=1
/label=MBP
/note="maltose binding protein from E. coli"
/translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
WYAVRTAVINAASGRQTVDEALKDAQT"
misc_feature 2633..2688
/gene="malE"
/label=protein spacer region
/note="protein spacer region"
CDS 2689..2709
/codon_start=1
/label=TEV site
/note="tobacco etch virus (TEV) protease recognition and
cleavage site"
/translation="ENLYFQG"
misc_feature 2711..2810
/gene="malE"
/label=multiple cloning site
/note="multiple cloning site"
primer_bind complement(2814..2830)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
terminator 3179..3265
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 3357..3384
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 3404..3495
/label=AmpR promoter
CDS 3496..4353
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin complement(4398..4911)
/direction=LEFT
/label=M13 ori
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
rep_origin 5022..5610
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(5796..5936)
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(6041..6229)
/codon_start=1
/label=rop
/note="Rop protein, which maintains plasmids at low copy
number"
/translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
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