Basic Vector Information
| Basic Vector Information | |||
|---|---|---|---|
| Vector Name | pUC-GM-INT | Antibiotic Resistance | Ampicillin |
| Length | 5646 bp | Type | Cloning vector |
| Source | Schweizer HD. | ||
pUC-GM-INT 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 ℃.
pUC-GM-INT vector Sequence
LOCUS Exported 5646 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pUC-GM-INT, complete sequence.
ACCESSION AF025392
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5646)
AUTHORS Schweizer HD.
TITLE Small broad-host-range gentamycin resistance gene cassettes for
site-specific insertion and deletion mutagenesis
JOURNAL BioTechniques 15 (5), 831-834 (1993)
PUBMED 8267974
REFERENCE 2 (bases 1 to 5646)
AUTHORS Mahenthiralingam E, Marklund BI, Brooks LA, Smith DA, Bancroft GJ,
Stokes RW.
TITLE Site-directed mutagenesis of the 19-kilodalton lipoprotein antigen
reveals No essential role for the protein in the growth and
virulence of Mycobacterium intracellulare
JOURNAL Infect. Immun. 66 (8), 3626-3634 (1998)
PUBMED 9673242
REFERENCE 3 (bases 1 to 5646)
AUTHORS Mahenthiralingam E, Marklund B-I., Brooks LA, Smith DA, Bancroft GJ,
Stokes RW.
TITLE Direct Submission
JOURNAL Submitted (16-SEP-1997) Pediatrics, University of British Columbia,
950 West 28th Avenue, Vancouver, BC V5Z 4H4, Canada
REFERENCE 4 (bases 1 to 5646)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..5646
/organism="Cloning vector pUC-GM-INT"
/mol_type="genomic DNA"
/note="L5-integrase based positive selection mycobacterial
cloning cassette"
/db_xref="taxon:67802"
primer_bind 379..395
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 453..481
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
CDS 670..1203
/codon_start=1
/gene="aacC1"
/product="gentamycin acetyl transferase"
/function="gentamycin-resistance"
/label=aacC1
/protein_id="AAC31620.1"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
gene 670..1203
/gene="aacC1"
/label=aacC1
misc_feature 1662..1740
/note="encodes mycobacteriophage attachment site, attP
core"
CDS 1989..2987
/codon_start=1
/product="L5-integrase"
/label=L5-integrase
/note="mycobacteriophage integrase"
/protein_id="AAC31622.1"
/translation="MDAEAWLAGEKRLIEMETWTPPQDRAKKAAASAITLEEYTRKWLV
ERDLADGTRDLYSGHAERRIYPVLGEVAVTEMTPALVRAWWAGMGRKHPTARRHAYNVL
RAVMNTAVEDKLIAENPCRIEQKAADERDVEALTPEELDIVAAEIFEHYRIAAYILAWT
SLRFGELIELRRKDIVDDGMTMKLRVRRGASRVGNKIVVGNAKTVRSKRPVTVPPHVAE
MIRAHMKDRTKMNKGPEAFLVTTTQGNRLSKSAFTKSLKRGYAKIGRPELRIHDLRAVG
ATFAAQAGATTKELMARLGHTTPRMAMKYQMASEARDEAIAEAMSKLAKTS"
primer_bind complement(3425..3441)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3449..3465
/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(3473..3503)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3518..3539
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3827..4415)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4586..5446)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=bla
/protein_id="AAC31621.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
gene complement(4586..5446)
/gene="bla"
/label=bla
promoter complement(5447..5551)
/gene="bla"
/label=AmpR promoter
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