pEM003 vector (V015311)

Basic Vector Information

Vector Name:
pEM003
Antibiotic Resistance:
Kanamycin
Length:
6447 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Mancera E, Frazer C, Porman AM, Ruiz-Castro S
Promoter:
lac

pEM003 vector Map

pEM0036447 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300CAP binding sitelac promoterlac operatorM13 revSP6 promoterARG4T7 promoterM13 fwdccdBNeoR/KanRBleoRori

pEM003 vector Sequence

LOCUS       62056_9145        6447 bp DNA     circular SYN 14-APR-2019
DEFINITION  Cloning vector pEM003, complete sequence.
ACCESSION   MK425747
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6447)
  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 6447)
  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 6447)
  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..6447
                     /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"
     gene            337..3264
                     /gene="ARG4"
                     /label=ARG4
                     /note="derived from Candida albicans"
     misc_feature    337..1321
                     /gene="ARG4"
                     /label=contains promoter and 5' UTR
                     /note="contains promoter and 5' UTR"
     CDS             1322..2728
                     /codon_start=1
                     /transl_table=11
                     /gene="ARG4"
                     /product="argininosuccinate lyase"
                     /label=ARG4
                     /protein_id="QBY25889.1"
                     /translation="MSQQQDKQPSENKLWGGRFTGATDPLMDLYNASLPYDKVMYDADL
                     TGTKVYTQGLNKLGLITTEELHLIHQGLEQIRQEWHDNKFIIKAGDEDIHTANERRLGE
                     IIGKNISGKVHTGRSRNDQVATDMRIFVRESLLNLSKILHQFITAILERAHKEIDVLMP
                     GYTHLQKAQPIRWAHWLSSYATYFTEDYKRLQEIITRVNQSPLGSGALAGHPYGIDREF
                     LAKGLGFDGVIGNSLTAVSDRDFVVESLFWSTLFMNHISRFSEDLIIYSSGEFGFIKLA
                     DAYSTGSSLMPQKKNPDSLELLRGKSGRVFGQLSGFLMSIKSIPSTYNKDMQEDKEPLF
                     DALTTVEHSILIATGVISTLLIDKQNMEKALTMDMLATDLADYLVRKGVPFRETHHISG
                     ECVRKAEEEKLSGIDQLSFEQFQQIDSRFEKDVMETFDFEASVERRDAIGGTAKSAVLK
                     QLENLKSILS"
     misc_feature    2729..3264
                     /gene="ARG4"
                     /label=contains 3' UTR and terminator
                     /note="contains 3' UTR and terminator"
     promoter        complement(3335..3353)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3360..3376)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3514..3813
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     CDS             4165..4956
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             5166..5537
                     /label=BleoR
                     /note="antibiotic-binding protein"
     rep_origin      5678..6266
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"

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