Basic Vector Information
- Vector Name:
- pJQ200mp18
- Antibiotic Resistance:
- Gentamycin
- Length:
- 5871 bp
- Type:
- Gene knockout
- Replication origin:
- p15A ori
- Host:
- E. coli
- Selection Marker:
- SacB
- Promoter:
- sacB
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
pJQ200mp18 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 ℃.
pJQ200mp18 vector Sequence
LOCUS 62056_13215 5871 bp DNA circular SYN 01-JAN-1980
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5871)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..5871
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter complement(491..519)
/label=Pc promoter
/note="class 1 integron promoter"
protein_bind 1347..1368
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 1383..1413
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1421..1437
/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 1445..1461
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 1470..1526
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(1530..1546)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin complement(2267..2812)
/direction=LEFT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin."
oriT complement(3195..3304)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
promoter 3680..4125
/label=sacB promoter
/note="sacB promoter and control region"
CDS 4126..5544
/codon_start=1
/label=SacB
/note="secreted levansucrase that renders bacterial growth
sensitive to sucrose"
/translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
VKDSILEQGQLTVNK"
CDS complement(join(5643..5871,1..302))
/codon_start=1
/label=GmR
/note="gentamycin acetyltransferase"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
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