Basic Vector Information
- Vector Name:
- pEM002
- Antibiotic Resistance:
- Kanamycin
- Length:
- 6176 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Mancera E, Frazer C, Porman AM, Ruiz-Castro S
pEM002 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 ℃.
pEM002 vector Sequence
LOCUS 62056_9140 6176 bp DNA circular SYN 14-APR-2019
DEFINITION Cloning vector pEM002, complete sequence.
ACCESSION MK425746
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6176)
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 6176)
AUTHORS Mancera E, Frazer C, Porman AM, Ruiz S, Johnson AD, Bennett RJ.
TITLE Direct Submission
JOURNAL Submitted (22-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 6176)
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
(22-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..6176
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 107..128
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/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 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 239..257
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
misc_feature 337..1322
/gene="LEU2"
/label=contains promoter and 5' UTR
/note="contains promoter and 5' UTR"
CDS 1323..2441
/gene="LEU2"
/label=LEU2
/note="3-isopropylmalate dehydrogenase from Candida
albicans. Accession#: P87186"
misc_feature 2445..2993
/gene="LEU2"
/label=contains 3' UTR and terminator
/note="contains 3' UTR and terminator"
promoter complement(3064..3082)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3089..3105)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
CDS 3243..3542
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
CDS 3894..4685
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
CDS 4895..5266
/label=BleoR
/note="antibiotic-binding protein"
rep_origin 5407..5995
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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