pEM002 vector (V015969)

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

pEM0026176 bp30060090012001500180021002400270030003300360039004200450048005100540057006000CAP binding sitelac promoterlac operatorM13 revSP6 promotercontains promoter and 5' UTR3-isopropylmalate dehydrogenasecontains 3' UTR and terminatorT7 promoterM13 fwdccdBNeoR/KanRBleoRori

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

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V015969                 6176 bp    DNA     circular SYN 14-APR-2019
DEFINITION  Exported.
ACCESSION   V015969
VERSION     V015969
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)"
            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="3-isopropylmalate dehydrogenase"
                     /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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