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 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 ℃.
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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