Basic Vector Information
- Vector Name:
- pME8
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7235 bp
- Type:
- Shuttle vector
- Replication origin:
- ori
- Source/Author:
- Audtho M, Ratlertkarn M, Wiwat C.
- Promoter:
- T3
pME8 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 ℃.
pME8 vector Sequence
LOCUS 40924_30575 7235 bp DNA circular SYN 18-DEC-2018
DEFINITION Shuttle vector pME8, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7235)
AUTHORS Audtho M, Ratlertkarn M, Wiwat C.
TITLE pME8, an improved shuttle vector for expression in Bacillus spp. and
E. coli
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7235)
AUTHORS Audtho M, Ratlertkarn M, Wiwat C.
TITLE Direct Submission
JOURNAL Submitted (21-FEB-2013) National Center for Genetic Engineering and
Biotechnology, National Science and Technology Development Agency,
113 Phaholyothin Road, Klong Nueng, Klong Luang, Pathum Thani 12120,
Thailand
REFERENCE 3 (bases 1 to 7235)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7235)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(21-FEB-2013) National Center for Genetic Engineering and
Biotechnology, National Science and Technology Development Agency,
113 Phaholyothin Road, Klong Nueng, Klong Luang, Pathum Thani 12120,
Thailand"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..7235
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 7..105
/note="contains cloning sites SmaI, PstI, EcoRI, EcoRV,
HindIII, BglII, SalI, NotI, SacI, SacII, PstI, XbaI, XhoI,
ApaI, and KpnI"
promoter complement(118..136)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(157..173)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(181..197)
/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)."
promoter complement(205..235)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(250..271)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(559..1147)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1321..2178)
/label=AmpR
/note="beta-lactamase"
promoter complement(2179..2283)
/label=AmpR promoter
rep_origin 2310..2765
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 2906..2922
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 2929..2947
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_difference 3389
/replace="g"
/label=G > C to delete EcoRI site in pBC16
/note="G > C to delete EcoRI site in pBC16"
CDS 3807..4808
/label=repB
/note="RepB replication protein"
CDS 5011..6384
/label=TcR
/note="tetracycline efflux protein"
misc_difference 6445
/replace="g"
/note="G > C to delete EcoRI site in DNA fragment from
pBC16"
rep_origin 6504..6780
/label=ori L
/note="ori L"
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