Basic Vector Information
pUCGM-lox 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 ℃.
pUCGM-lox vector Sequence
LOCUS 40924_45258 3695 bp DNA circular SYN 18-DEC-2018
DEFINITION Gene deletion vector pUCGM-lox, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3695)
AUTHORS Quenee L, Lamotte D, Polack B.
TITLE Combined sacB-based negative selection and cre-lox antibiotic marker
recycling for efficient gene deletion in pseudomonas aeruginosa
JOURNAL BioTechniques 38 (1), 63-67 (2005)
PUBMED 15679087
REFERENCE 2 (bases 1 to 3695)
AUTHORS Quenee L, Lamotte D, Polack B.
TITLE Direct Submission
JOURNAL Submitted (27-JAN-2005) GREPI, CHU, Hopital Michallon, Grenoble
38043, France
REFERENCE 3 (bases 1 to 3695)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3695)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"BioTechniques"; date: "2005"; volume: "38"; issue: "1"; pages:
"63-67"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(27-JAN-2005) GREPI, CHU, Hopital Michallon, Grenoble 38043, France"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..3695
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 379..395
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 492..525
/label=Cre recombinase binding site
/bound_moiety="Cre recombinase"
/note="loxP, Cre recombinase recognition site"
promoter 534..562
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
CDS 751..1281
/codon_start=1
/label=GmR
/note="gentamycin acetyltransferase"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
protein_bind complement(1328..1361)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
primer_bind complement(1474..1490)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(1498..1514)
/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(1522..1552)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(1567..1588)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(1876..2464)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2638..3495)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(3496..3600)
/label=AmpR promoter
This page is informational only.