Basic Vector Information
- Vector Name:
- pP01680-nat-cel8A
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6806 bp
- Type:
- Shuttle vector
- Replication origin:
- ori
- Source/Author:
- Heinze S, Zimmermann K, Ludwig C, Heinzlmeir S, Schwarz WH, Zverlov VV, Liebl W, Kornberger P.
pP01680-nat-cel8A 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 ℃.
pP01680-nat-cel8A vector Sequence
LOCUS 40924_33977 6806 bp DNA circular SYN 18-DEC-2018
DEFINITION Shuttle vector pP01680-nat-cel8A, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6806)
AUTHORS Heinze S, Zimmermann K, Ludwig C, Heinzlmeir S, Schwarz WH, Zverlov
VV, Liebl W, Kornberger P.
TITLE Efficient secretory heterologous enzyme production in the novel host
Paenibacillus polymyxa by usage of its promoter sequences
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6806)
AUTHORS Heinze S, Zimmermann K, Ludwig C, Heinzlmeir S, Schwarz WH, Zverlov
VV, Liebl W, Kornberger P.
TITLE Direct Submission
JOURNAL Submitted (09-MAY-2018) Department for Microbiology, Technical
University of Munich, Emil-Ramann-Str. 4, Freising, Bavaria 85354,
Germany
REFERENCE 3 (bases 1 to 6806)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6806)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(09-MAY-2018) Department for Microbiology, Technical University of
Munich, Emil-Ramann-Str. 4, Freising, Bavaria 85354, Germany"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..6806
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(79..667)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(841..1698)
/label=AmpR
/note="beta-lactamase"
promoter complement(1699..1803)
/label=AmpR promoter
CDS complement(1832..2116)
/codon_start=1
/gene="traJ"
/product="TraJ"
/label=traJ
/protein_id="AXF54520.1"
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
GQGYKITGVVDYEHVRELARINGDLGQQKAHYVNHHPNQVFWGRGAVKH"
gene complement(1832..2116)
/gene="traJ"
/label=traJ
oriT complement(2149..2258)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
CDS complement(2515..3276)
/codon_start=1
/gene="kanR"
/product="KanR"
/label=kanR
/note="Kanamycin Resistance"
/protein_id="AXF54521.1"
/translation="MNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGRQTD
GPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFDSEEILLDYASQVESDWPLTHGQFF
SILPIYDSGGYLEKVYQTAKSVEAQTFHDAICALIVEELFEYAGKWRNIRVQGPTTFLP
SLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQSDLPSGYDHLCQFVMSGQLSDSEK
LLESLENFWNGIQEWTERHGYIVDVSKRIPF"
gene complement(2515..3276)
/gene="kanR"
/label=kanR
CDS complement(3448..4449)
/label=repB
/note="RepB replication protein"
regulatory 4866..5158
/label=P01680 promoter from P. polymyxa DSM292
/note="P01680 promoter from P. polymyxa DSM292"
/regulatory_class="promoter"
CDS 5156..5267
/codon_start=1
/gene="SPnat-Cel8A-His6"
/product="signal peptide of P. polymyxa DSM292 ORF
PPOLYM_01680"
/label=SPnat-Cel8A-His6
/protein_id="AXF54522.1"
/translation="MKSFLVKSLLALSVLFMLLGFMPVGQASVSANSVGAE"
CDS 6626..6643
/label=6xHis
/note="6xHis affinity tag"
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