Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012554 | pAN7-1 | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pAN7-1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6760 bp
- Type:
- Cloning Vectors
- Replication origin:
- ori
- Source/Author:
- Punt PJ, Oliver RP, Dingemanse MA, Pouwels PH, van den Hondel CA.
- Copy Number:
- High copy number
- Promoter:
- gpdA
- Growth Strain(s):
- DH5alpha
- Growth Temperature:
- 37℃
pAN7-1 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.
pAN7-1 vector Sequence
LOCUS Exported 6760 bp DNA circular SYN 03-SEP-2024
DEFINITION Vector with a hygromycin B resistance marker for transforming
Aspergillus species.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6760)
AUTHORS Punt PJ, Oliver RP, Dingemanse MA, Pouwels PH, van den Hondel CA.
TITLE Transformation of Aspergillus based on the hygromycin B resistance
marker from Escherichia coli.
JOURNAL Gene 1987;56:117-24.
PUBMED 2824287
REFERENCE 2 (bases 1 to 6760)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6760)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6760)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Gene";
date: "1987"; volume: "56"; pages: "117-24"
COMMENT SGRef: number: 2; type: "Journal Article"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT The trpC terminator region was corrected based on the updated
sequence record X02390.
FEATURES Location/Qualifiers
source 1..6760
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..2129
/label=gpdA promoter
/note="promoter from the Aspergillus nidulans
glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
1990)"
intron 2178..2293
/label=gpdA intron
/note="intron from the Aspergillus nidulans
glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
1990)"
CDS 2302..3318
/codon_start=1
/label=HygR
/note="aminoglycoside phosphotransferase from E. coli"
/translation="MPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
terminator 3337..4112
/label=trpC terminator
/note="transcription terminator from the Aspergillus
nidulans trpC gene (Punt et al., 1987)"
primer_bind complement(4135..4151)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 4625..4729
/label=AmpR promoter
CDS 4730..5587
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 5761..6349
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 6637..6658
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6673..6703
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6711..6727
/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 6735..6751
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"