pYEp352 Catalog No. PVT11240
Packing 2ug
pYEp352 Information
Function yeast expression plasmid
romoter: URA3
Replicator: 2 ori, ori
Plasmid classification: yeast series, Saccharomyces cerevisiae expression vector
Plasmid size: 5881bp
Prokaryotic resistance: Amp
Screening markers: URA3
Cloned strain: DH5 alpha
Culture conditions: 37 centigrade, aerobic LB
Expression host: yeast cells
5'sequencing primers: M13F:TGTAAAACGACGGCCAGT
3'sequencing primers: primers designed according to sequence
pYEp352 Description
Restriction digests of the clone give the following sizes (kb): BamHI--5.3; XbaI--5.3; PstI--5.3; HindIII--5.3.This is a Escherichia coli/yeast episomal shuttle vector with multiple restriction sites for cloning.Constructed by filling in and blunt-end ligating pUC18 linearized at NdeI, a 1.4 kb RsaI fragment of the 2 micron circle, and a 1.1 kb HindIII/SmaI fragment containing URA3. A XbaI site in 2u and a PstI site in URA3 were removed.The yeast genomicsequence is in nt 286-1383 (where the A of the ATG of lacZ' is number 1), the 2 micron sequence goes from nt 1384-2768, and pUC18 includes nt 2769-285.
pYEp352 Sequence
LOCUS Exported 5181 bp ds-DNA circular SYN 11-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 5
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5181)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sunday, September 11, 2016 from SnapGene Viewer 3.1.4
FEATURES Location/Qualifiers
source 1..5181
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 20..36
/bound_moiety="lac repressor encoded by lacI"
/note="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(44..74)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
rep_origin complement(398..986)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1157..2017)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(2018..2122)
/gene="bla"
/note="AmpR promoter"
rep_origin 2404..3568
/note="2u ori"
/note="yeast 2u plasmid origin of replication"
promoter 3804..4014
/gene="S. cerevisiae URA3"
/note="URA3 promoter"
CDS 4015..4818
/codon_start=1
/gene="S. cerevisiae URA3"
/product="orotidine-5'-phosphate decarboxylase, required
for uracil biosynthesis"
/note="URA3"
/note="yeast auxotrophic marker, counterselectable with
5-fluoroorotic acid (5-FOA)"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
CDS complement(4891..5181)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/note="lacZ-alpha"
/translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIG"
primer_bind 5091..5107
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 5111..5167
/note="MCS"
/note="pUC18/19 multiple cloning site"
primer_bind complement(5177..12)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
61 gcataaagtg taaagcctgg ggtgcctaat gagtgaggta actcacatta attgcgttgc
121 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctggattaa tgaatcggcc
181 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
241 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
301 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
361 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
421 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
481 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
541 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac
601 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
661 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
721 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
781 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
841 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
901 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
961 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
1021 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
1081 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
温馨提示：不可用于临床ZL。
