Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V017514 | pSFS2A | 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:
- pSFS2A
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 7510 bp
- Copy Number:
- High copy number
- Growth Strain(s):
- DH5alpha
- Growth Temperature:
- 37℃
pSFS2A 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.
pSFS2A vector Sequence
LOCUS Exported 7510 bp DNA circular SYN 23-DEC-2024
DEFINITION Cloning vector pSFS2A-mNeonGreen, complete sequence.
ACCESSION MK431400
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7510)
AUTHORS Mancera E, Frazer C, Porman AM, Ruiz-Castro S, Johnson AD, Bennett
RJ.
TITLE Genetic Modification of Closely Related Candida Species
JOURNAL Front Microbiol 10, 357 (2019)
PUBMED 30941104
REFERENCE 2 (bases 1 to 7510)
AUTHORS Mancera E, Frazer C, Porman AM, Ruiz S, Johnson AD, Bennett RJ.
TITLE Direct Submission
JOURNAL Submitted (23-JAN-2019) Departmento de Ingenieria Genetica, Centro
de Investigacion y de Estudios Avanzados del Instituto Politecnico
Nacional, Unidad Irapuato, Km. 9.6 Libramiento Norte Carrera
Irapuato-Leon, Irapuato, Guanajuato 36824, Mexico
REFERENCE 3 (bases 1 to 7510)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7510)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Front
Microbiol 10, 357 (2019)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(23-JAN-2019) Departmento de Ingenieria Genetica, Centro de
Investigacion y de Estudios Avanzados del Instituto Politecnico
Nacional, Unidad Irapuato, Km. 9.6 Libramiento Norte Carrera
Irapuato-Leon, Irapuato, Guanajuato 36824, Mexico"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7510
/mol_type="other DNA"
/organism="synthetic DNA construct"
source join(915..7510,1..914)
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter complement(142..246)
/label=AmpR promoter
rep_origin 273..729
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 870..886
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 896..914
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind complement(944..977)
/label=FRT (minimal)
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
misc_feature 982..1529
/label=promoter region from Candida albicans MAL2
/note="promoter region from Candida albicans MAL2"
CDS 1536..2804
/label=FLP
/note="site-specific recombinase"
misc_feature 2808..3195
/label=terminator region from Candida albicans ACT1
/note="terminator region from Candida albicans ACT1"
misc_feature 3202..3699
/label=promoter region from Candida albicans ACT1
/note="promoter region from Candida albicans ACT1"
gene 3700..4926
/gene="SAT1"
/label=SAT1
CDS join(3700..3709,4364..4926)
/codon_start=1
/transl_table=11
/gene="SAT1"
/product="streptomycin acetyltransferase"
/label=SAT1
/note="nourseothricin resistance marker"
/protein_id="QBY25799.1"
/translation="MDGEEVAALVIDNGSHMKISVIPEQVAETLDAENHFIVREVFDVH
LSDQGFELSTRSVSPYRKDYISDDDSDEDSACYGAFIDQELVGKIELNSTWNDLASIEH
IVVSHTHRGKGVAHSLIEFAKKWALSRQLLGIRLETQTNNVPACNLYAKCGFTLGGIDL
FTYKTRPQVSNETAMYWYWFSGAQDDA"
misc_feature 4930..5059
/label=terminator region from Candida albicans URA3
/note="terminator region from Candida albicans URA3"
protein_bind complement(5064..5097)
/label=FRT (minimal)
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
primer_bind complement(5099..5115)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(5152..5170)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(5191..5207)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(5215..5231)
/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(5239..5269)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(5284..5305)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5593..6181)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 6401..6503
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS 6504..7160
/label=CmR
/note="chloramphenicol acetyltransferase"