Basic Vector Information
- Vector Name:
- pMAL-pIII
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6706 bp
- Type:
- Shuttle vector
- Replication origin:
- ori
- Source/Author:
- Zwick MB, Bonnycastle LL, Noren KA, Venturini S, Leong E, Barbas CF III, Noren CJ, Scott JK.
pMAL-pIII 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 ℃.
pMAL-pIII vector Sequence
LOCUS 40924_29766 6706 bp DNA circular SYN 18-DEC-2018
DEFINITION Shuttle vector pMAL-pIII, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6706)
AUTHORS Zwick MB, Bonnycastle LL, Noren KA, Venturini S, Leong E, Barbas CF
III, Noren CJ, Scott JK.
TITLE The maltose-binding protein as a scaffold for monovalent display of
peptides derived from phage libraries
JOURNAL Anal. Biochem. 264 (1), 87-97 (1998)
PUBMED 9784192
REFERENCE 2 (bases 1 to 6706)
AUTHORS Noren KA, Noren CJ.
TITLE Direct Submission
JOURNAL Submitted (22-OCT-1997) New England Biolabs, 32 Tozer Road, Beverly,
MA 01915, USA
REFERENCE 3 (bases 1 to 6706)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6706)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Anal.
Biochem."; date: "1998"; volume: "264"; issue: "1"; pages: "87-97"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(22-OCT-1997) New England Biolabs, 32 Tozer Road, Beverly, MA 01915,
USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6706
/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 1534..2688
/codon_start=1
/label=MBP
/note="maltose binding protein from E. coli"
/translation="KLLFAIPLVVPFYSHSADMKIEEGKLVIWINGDKGYNGLAEVGKK
FEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAF
QDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGK
SALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKH
MNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVL
SAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATM
ENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQT"
CDS 2737..2748
/codon_start=1
/label=Factor Xa site
/note="Factor Xa recognition and cleavage site"
/translation="IEGR"
primer_bind complement(2796..2812)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
terminator 3161..3247
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 3339..3366
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 3386..3477
/label=AmpR promoter
CDS 3478..4335
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin complement(4380..4893)
/direction=LEFT
/label=M13 ori
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
rep_origin 5004..5592
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(5778..5918)
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(6023..6211)
/codon_start=1
/label=rop
/note="Rop protein, which maintains plasmids at low copy
number"
/translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
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