Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V001816 | pUCmini-iCAP-PHP.eB | 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:
- pUCmini-iCAP-PHP.eB
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8543 bp
- Type:
- Mammalian Expression, AAV
- Replication origin:
- ori
- Promoter:
- p41
- Cloning Method:
- Gibson Cloning
pUCmini-iCAP-PHP.eB 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.
pUCmini-iCAP-PHP.eB vector Sequence
LOCUS V001816 8543 bp DNA circular SYN 12-JUN-2021
DEFINITION Exported.
ACCESSION V001816
VERSION V001816
KEYWORDS pUCmini-iCAP-PHP.eB
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 8543)
AUTHORS Chan KY, Jang MJ, Yoo BB, Greenbaum A, Ravi N, Wu WL,
Sanchez-Guardado L, Lois C, Mazmanian SK, Deverman BE, Gradinaru V
TITLE Engineered AAVs for efficient noninvasive gene delivery to the
central and peripheral nervous systems.
JOURNAL Nat Neurosci. 2017 Aug;20(8):1172-1179. doi: 10.1038/nn.4593. Epub
2017 Jun 26.
PUBMED 28671695
REFERENCE 2 (bases 1 to 8543)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 8543)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nn.4593";
journalName: "Nat Neurosci"; date: "2017-08"; volume: "20"; issue:
"8"; pages: "1172-1179"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8543
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 114..130
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 140..158
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 239..2101
/gene="Rep78"
/label="Protein Rep78"
/note="Protein Rep78 from Adeno-associated virus 2 (isolate
Srivastava/1982). Accession#: Q89268"
CDS 2121..2864
/label="rtTA-Advanced"
/note="improved tetracycline-controlled transactivator"
promoter complement(2897..2915)
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
polyA_signal 2941..3165
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
protein_bind 3227..3245
/label="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3264..3282
/gene="tetO"
/label="tet operator"
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3301..3319
/gene="tetO"
/label="tet operator"
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3338..3356
/gene="tetO"
/label="tet operator"
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3375..3393
/gene="tetO"
/label="tet operator"
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3412..3430
/gene="tetO"
/label="tet operator"
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3449..3467
/gene="tetO"
/label="tet operator"
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3486..3504
/gene="tetO"
/label="tet operator"
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
CDS 6221..6232
/label="WELQut site"
/note="WELQut protease recognition and cleavage site"
rep_origin complement(6788..7376)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7550..8407)
/label="AmpR"
/note="beta-lactamase"
promoter complement(8408..8512)
/label="AmpR promoter"