Basic Vector Information
- Vector Name:
- pEM008
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 7944 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Mancera E, Frazer C, Porman AM, Ruiz-Castro S
pEM008 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 ℃.
pEM008 vector Sequence
LOCUS 62056_9155 7944 bp DNA circular SYN 14-APR-2019
DEFINITION Cloning vector pEM008, complete sequence.
ACCESSION MK431394
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7944)
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 7944)
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 7944)
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"
FEATURES Location/Qualifiers
source 1..7944
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_recomb 22..55
/label=FLP recombination target sequence (FRT)
/note="FLP recombination target sequence (FRT)"
misc_feature 62..1048
/label=promoter region from Candida tropicalis PCK1
/note="promoter region from Candida tropicalis PCK1"
CDS 1049..2317
/label=FLP
/note="site-specific recombinase"
misc_feature 2321..2708
/label=terminator region from Candida albicans ACT1
/note="terminator region from Candida albicans ACT1"
misc_feature 2715..3212
/label=promoter region from Candida albicans ACT1
/note="promoter region from Candida albicans ACT1"
gene 3213..4439
/gene="SAT1"
/label=SAT1
CDS join(3213..3222,3877..4439)
/codon_start=1
/transl_table=11
/gene="SAT1"
/product="streptomycin acetyltransferase"
/label=SAT1
/note="nourseothricin resistance marker"
/protein_id="QBY25788.1"
/translation="MDGEEVAALVIDNGSHMKISVIPEQVAETLDAENHFIVREVFDVH
LSDQGFELSTRSVSPYRKDYISDDDSDEDSACYGAFIDQELVGKIELNSTWNDLASIEH
IVVSHTHRGKGVAHSLIEFAKKWALSRQLLGIRLETQTNNVPACNLYAKCGFTLGGIDL
FTYKTRPQVSNETAMYWYWFSGAQDDA"
misc_feature 4443..4572
/label=terminator region from Candida albicans URA3
/note="terminator region from Candida albicans URA3"
protein_bind complement(4577..4610)
/label=FRT (minimal)
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
primer_bind complement(4612..4628)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(4665..4683)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(4704..4720)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4728..4744)
/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(4752..4782)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4797..4818)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5106..5694)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 5914..6016
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS 6017..6673
/label=CmR
/note="chloramphenicol acetyltransferase"
promoter complement(7164..7269)
/label=AmpR promoter
rep_origin 7296..7751
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 7892..7908
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 7918..7936
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
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