Basic Vector Information
- Vector Name:
- pPL5617_pUG_PuroR
- Antibiotic Resistance:
- Ampicillin
- Length:
- 3780 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Host:
- Yeast
- Source/Author:
- MacDonald C, Piper RC.
- Promoter:
- TEF
pPL5617_pUG_PuroR 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 ℃.
pPL5617_pUG_PuroR vector Sequence
LOCUS 40924_34889 3780 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pPL5617_pUG_PuroR, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3780)
AUTHORS MacDonald C, Piper RC.
TITLE Puromycin- and methotrexate-resistance cassettes and optimized
Cre-recombinase expression plasmids for use in yeast
JOURNAL Yeast 32 (5), 423-438 (2015)
PUBMED 25688547
REFERENCE 2 (bases 1 to 3780)
AUTHORS MacDonald C, Piper RC.
TITLE Direct Submission
JOURNAL
REFERENCE 3 (bases 1 to 3780)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3780)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Yeast";
date: "2015"; volume: "32"; issue: "5"; pages: "423-438"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(18-NOV-2014) Molecular Physiology "
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Assembly Method :: Sequence Builder (Lasergene) v. 8.0.3
Sequencing Technology :: Assembled
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..3780
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 53..86
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
promoter 141..484
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
CDS 485..1081
/codon_start=1
/label=PuroR
/note="puromycin N-acetyltransferase"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
terminator 1090..1287
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
misc_feature 1331..1364
/label=loxP site
/note="loxP site"
protein_bind complement(1331..1364)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
promoter complement(1418..1436)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
rep_origin complement(1694..2282)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2456..3313)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(3314..3418)
/label=AmpR promoter
promoter 3764..3780
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
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