Basic Vector Information
- Vector Name:
- pLoxGentrc
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6295 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Palmeros B, Wild J, Szybalski W, Le Borgne S, Hernandez-Chavez G, Gosset G, Valle F, Bolivar F.
pLoxGentrc vector Map
pLoxGentrc vector Sequence
LOCUS 40924_28606 6295 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pLoxGentrc, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6295)
AUTHORS Palmeros B, Wild J, Szybalski W, Le Borgne S, Hernandez-Chavez G,
Gosset G, Valle F, Bolivar F.
TITLE A family of removable cassettes designed to obtain
antibiotic-resistance-free genomic modifications of Escherichia coli
and other bacteria
JOURNAL Gene 247 (1-2), 255-264 (2000)
PUBMED 10773465
REFERENCE 2 (bases 1 to 6295)
AUTHORS Sabido A, Martinez LM, de Anda R, Martinez A, Bolivar F, Gosset G.
TITLE A novel plasmid vector designed for chromosomal gene integration and
expression: Use for developing a genetically stable Escherichia coli
melanin production strain
JOURNAL Plasmid 69 (1), 16-23 (2013)
PUBMED 22884755
REFERENCE 3 (bases 1 to 6295)
AUTHORS Sabido A, Gosset G.
TITLE Direct Submission
JOURNAL Submitted (11-MAY-2011) Cell Engineering and Biocatalysis,
Biotechnology Institute, Av. Universidad 2001, Cuernavaca, Morelos
62210, Mexico
REFERENCE 4 (bases 1 to 6295)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 6295)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Gene 247
(1-2), 255-264 (2000)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Plasmid";
date: "2013"; volume: "69"; issue: "1"; pages: "16-23"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(11-MAY-2011) Cell Engineering and Biocatalysis, Biotechnology
Institute, Av. Universidad 2001, Cuernavaca, Morelos 62210, Mexico"
COMMENT SGRef: number: 4; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6295
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(1..456)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 669..685
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 733..749
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 763..796
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
CDS complement(843..1373)
/label=GmR
/note="gentamycin acetyltransferase"
regulatory complement(1557..1563)
/gene="aacC1"
/regulatory_class="minus_10_signal"
promoter complement(1562..1590)
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
regulatory complement(1585..1590)
/gene="aacC1"
/regulatory_class="minus_35_signal"
misc_feature complement(1599..1632)
/note="loxP site; DNA target site for Cre site-specific
recombinase"
protein_bind complement(1599..1632)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
terminator complement(1716..1743)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(1835..1921)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
rRNA complement(1912..2041)
/gene="rrnB"
/product="5S ribosomal RNA"
/label=rrnB
misc_feature complement(2124..2180)
/label=MCS
/note="pUC18/19 multiple cloning site"
regulatory complement(2192..2195)
/label=from lacZ
/note="from lacZ"
/regulatory_class="ribosome_binding_site"
protein_bind 2204..2220
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(2228..2257)
/label=trc promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
CDS complement(2492..3571)
/label=lacI
/note="lac repressor"
promoter complement(3572..3649)
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
misc_feature complement(3650..3680)
/label=lacIq 5' sequence
/note="lacIq 5' sequence"
misc_feature complement(3681..3873)
/label=intergenic region from pTrc99A
/note="intergenic region from pTrc99A"
primer_bind complement(3888..3904)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3912..3928)
/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(3936..3966)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3981..4002)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4290..4878)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5052..5909)
/label=AmpR
/note="beta-lactamase"
promoter complement(5910..6014)
/label=AmpR promoter
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