Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V014802 | pTrp-GFA | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pTrp-GFA
- Antibiotic Resistance:
- Tetracycline
- Length:
- 17509 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- E. coli
- Promoter:
- Tac
- Growth Temperature:
- 37℃
pTrp-GFA vector Map
Plasmid 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 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
pTrp-GFA vector Sequence
LOCUS V014802 17509 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V014802
VERSION V014802
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 17509)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..17509
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 9..37
/label="tac promoter"
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
CDS 174..1223
/gene="aroG"
/label="Phospho-2-dehydro-3-deoxyheptonate aldolase,
Phe-sensitive"
/note="Phospho-2-dehydro-3-deoxyheptonate aldolase,
Phe-sensitive from Escherichia coli (strain K12).
Accession#: P0AB91"
CDS 1227..2294
/gene="aroF"
/label="Phospho-2-dehydro-3-deoxyheptonate aldolase,
Tyr-sensitive"
/note="Phospho-2-dehydro-3-deoxyheptonate aldolase,
Tyr-sensitive from Escherichia coli (strain K12).
Accession#: P00888"
CDS 2298..4286
/gene="tktA"
/label="Transketolase 1"
/note="Transketolase 1 from Escherichia coli (strain K12).
Accession#: P27302"
CDS 4704..5933
/gene="serA"
/label="D-3-phosphoglycerate dehydrogenase"
/note="D-3-phosphoglycerate dehydrogenase from Escherichia
coli (strain K12). Accession#: P0A9T0"
promoter 6040..6068
/label="tet promoter"
/note="E. coli promoter for tetracycline efflux protein
gene"
CDS 6116..7303
/label="TcR"
/note="tetracycline efflux protein"
CDS 7945..8133
/label="rop"
/note="Rop protein, which maintains plasmids at low copy
number"
misc_feature 8238..8378
/label="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(8564..9152)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 9299..9376
/label="lacI promoter"
promoter 10455..10483
/label="tac promoter"
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 10491..10507
/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 10515..10531
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
CDS 10613..12172
/gene="trpE"
/label="Anthranilate synthase component 1"
/note="Anthranilate synthase component 1 from Escherichia
coli (strain K12). Accession#: P00895"
CDS 12175..13767
/gene="trpGD"
/label="Bifunctional protein TrpGD"
/note="Bifunctional protein TrpGD from Escherichia coli
(strain K12). Accession#: P00904"
CDS 13771..15129
/gene="trpC"
/label="Tryptophan biosynthesis protein TrpCF"
/note="Tryptophan biosynthesis protein TrpCF from
Escherichia coli (strain K12). Accession#: P00909"
CDS 15144..16334
/gene="trpB"
/label="Tryptophan synthase beta chain"
/note="Tryptophan synthase beta chain from Shigella
flexneri. Accession#: P0A880"
CDS 16337..17140
/gene="trpA"
/label="Tryptophan synthase alpha chain"
/note="Tryptophan synthase alpha chain from Escherichia
coli (strain K12). Accession#: P0A877"