| 质粒名称: | pPL5071_TEF1*-Cre_URA3 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 60930 |
| 存储实验室: | Robert Piper |
|
载体骨架基本信息 |
|
| 载体名称: | pSH47 |
| 完整质粒大小: | 6906 bp |
| 载体类型: | Yeast Expression |
| 筛选标记: | URA3 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | Top10 |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Cre / Cre-recombinase |
| 物种来源: | E. coli |
| 插入大小: | 1032 bp |
| 启动子: | mutant TEF1 |
| 标签/融合蛋白: | Tag / Fusion Protein |
| 克隆方法: | 未知 |
| 5’测序引物 | cgataccgtcgaggggcagagcc |
| 3’测序引物 | catattttccgtcgatgttgaaatcc |
|
文献引用方法 |
|
| 材料和方法部分: | pPL5071_TEF1*-Cre_URA3 was a gift from Robert Piper (Addgene plasmid # 60930 ; http://n2t.net/addgene:60930 ; RRID:Addgene_60930) |
| 参考文献部分: | Puromycin and Methotrexate Resistance Cassettes and Optimized cre-recombinase Expression Plasmids for use in Yeast. MacDonald C, Piper RC. Yeast. 2015 Feb 16. doi: 10.1002/yea.3069. 10.1002/yea.3069 PubMed 25688547 |
买家导航
pPL5071_TEF1*-Cre_URA3载体质粒基本信息
pPL5071_TEF1*-Cre_URA3质粒图谱载体图谱和pPL5071_TEF1*-Cre_URA3载体序列质粒序列多克隆位点信息
pPL5071_TEF1*-Cre_URA3质粒载体简介
pPL5071_TEF1*-Cre_URA3质粒序列载体序列
LOCUS pPL5071_TEF1*-Cre_URA3 6906 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Constitutive expression vector for Cre-recombinase containing URA3
selection marker..
ACCESSION .
VERSION .
KEYWORDS pPL5071_TEF1*-Cre_URA3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6906)
AUTHORS MacDonald C, Piper RC
TITLE Puromycin and Methotrexate Resistance Cassettes and Optimized
cre-recombinase Expression Plasmids for use in Yeast.
JOURNAL Yeast. 2015 Feb 16. doi: 10.1002/yea.3069.
PUBMED 25688547
REFERENCE 2 (bases 1 to 6906)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6906
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..19
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
CDS 553..1584
/codon_start=1
/product="site-specific recombinase"
/label=Cre
/note="Cre is a site-specific recombinase from
bacteriophage P1. Recombination occurs at loxP sequences."
/translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
DGD"
terminator 1902..2149
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind complement(1918..1936)
/label=CYC1
/note="CYC1 transcription termination signal, reverse
primer"
primer_bind complement(2167..2186)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(2168..2186)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2196..2213)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(2196..2212)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2205..2227)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 2353..2808
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2440..2459)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2650..2671
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(2939..3742)
/codon_start=1
/gene="S. cerevisiae URA3"
/product="orotidine-5'-phosphate decarboxylase, required
for uracil biosynthesis"
/label=URA3
/note="yeast auxotrophic marker, counterselectable with
5-fluoroorotic acid (5-FOA)"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
promoter complement(3743..3958)
/gene="S. cerevisiae URA3"
/label=URA3 promoter
primer_bind complement(3982..4001)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4101..4123
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4161..4179)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
misc_feature 4220..4723
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 4760..4864
/gene="bla"
/label=AmpR promoter
CDS 4865..5725
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(5083..5102)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5896..6484
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6385..6404
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6638..6655
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 6772..6793
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6808..6838
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6846..6862
/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)."
primer_bind 6851..6873
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 6870..6886
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 6870..6886
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 6905..19
/label=T3
/note="T3 promoter, forward primer"
ORIGIN
1 aattaaccct cactaaaggg aacaaaagct ggatagcttc aaaatgtctc tactcctttt
61 ttactcttcc agattttctc ggactccgcg caccgccgta ccacttcaaa acacccaagc
121 acagcatact aaattccccc tcctccttcc tctagggtgc cgttaattac ccgtactaaa
181 ggtttggaaa aggaaaaaga gaccgcctcg tccctttttc ttcgtcggag aaggcaataa
241 aaatttttat cacgtttctt tctcttgaaa actttttttt cgattttgtt ctctttcgac
301 gacctcccat tgatatttga gttaacaaac ggtcttcaat ttctcaagtt tcagcttcat
361 ttttcctgtt ctattacaac tttttttact tcttgctcat tggaaagaaa gcatagcaat
421 ctaatctaag ttttaattac aaaaactagt ggatcccccg ggctgcagga attcgatatc
481 aagcttatcg ataccgtcga ggggcagagc cgatcctgta cactttactt aaaaccatta
541 tctgagtgtt aaatgtccaa tttactgacc gtacaccaaa atttgcctgc attaccggtc
601 gatgcaacga gtgatgaggt tcgcaagaac ctgatggaca tgttcaggga tcgccaggcg
661 ttttctgagc atacctggaa aatgcttctg tccgtttgcc ggtcgtgggc ggcatggtgc
721 aagttgaata accggaaatg gtttcccgca gaacctgaag atgttcgcga ttatcttcta
781 tatcttcagg cgcgcggtct ggcagtaaaa actatccagc aacatttggg ccagctaaac
841 atgcttcatc gtcggtccgg gctgccacga ccaagtgaca gcaatgctgt ttcactggtt
901 atgcggcgga tccgaaaaga aaacgttgat gccggtgaac gtgcaaaaca ggctctagcg
961 ttcgaacgca ctgatttcga ccaggttcgt tcactcatgg aaaatagcga tcgctgccag
1021 gatatacgta atctggcatt tctggggatt gcttataaca ccctgttacg tatagccgaa
1081 attgccagga tcagggttaa agatatctca cgtactgacg gtgggagaat gttaatccat
1141 attggcagaa cgaaaacgct ggttagcacc gcaggtgtag agaaggcact tagcctgggg
1201 gtaactaaac tggtcgagcg atggatttcc gtctctggtg tagctgatga tccgaataac
1261 tacctgtttt gccgggtcag aaaaaatggt gttgccgcgc catctgccac cagccagcta
1321 tcaactcgcg ccctggaagg gatttttgaa gcaactcatc gattgattta cggcgctaag
1381 gatgactctg gtcagagata cctggcctgg tctggacaca gtgcccgtgt cggagccgcg
1441 cgagatatgg cccgcgctgg agtttcaata ccggagatca tgcaagctgg tggctggacc
1501 aatgtaaata ttgtcatgaa ctatatccgt aacctggata gtgaaacagg ggcaatggtg
1561 cgcctgctgg aagatggcga ttagccatta acgcgtaaat gattgctata attatttgat
1621 atttatggtg acatatgaga aaggatttca acatcgacgg aaaatatgta gtgctgtctg
1681 taagcactaa tattcagtcg ccagccgtca ttgtcactgt aaagctgagc gatagaatgc
1741 ctgatattga ctcaatatcc gttgcgtttc ctgtcaaaag tatgcgtagt gctgaacatt
1801 tcgtgatgaa tgccaccgag gaagaagcac ggcgcggttt tgctaaagtg atgtctgagt
1861 ttggcgaact cttgggtaag gttggaattg tcgacctcga gtcatgtaat tagttatgtc
1921 acgcttacat tcacgccctc cccccacatc cgctctaacc gaaaaggaag gagttagaca
1981 acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa gaacgttatt
2041 tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta acattatact
2101 gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgcg gccggtaccc
2161 aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt
2221 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
2281 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
2341 aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
2401 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
2461 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
2521 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
2581 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
2641 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
2701 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
2761 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta
2821 ttttctcctt acgcatctgt gcggtatttc acaccgcata gggtaataac tgatataatt
2881 aaattgaagc tctaatttgt gagtttagta tacatgcatt tacttataat acagtttttt
2941 agttttgctg gccgcatctt ctcaaatatg cttcccagcc tgcttttctg taacgttcac
3001 cctctacctt agcatccctt ccctttgcaa atagtcctct tccaacaata ataatgtcag
3061 atcctgtaga gaccacatca tccacggttc tatactgttg acccaatgcg tctcccttgt
3121 catctaaacc cacaccgggt gtcataatca accaatcgta accttcatct cttccaccca
3181 tgtctctttg agcaataaag ccgataacaa aatctttgtc gctcttcgca atgtcaacag
3241 tacccttagt atattctcca gtagataggg agcccttgca tgacaattct gctaacatca
3301 aaaggcctct aggttccttt gttacttctt ctgccgcctg cttcaaaccg ctaacaatac
3361 ctgggcccac cacaccgtgt gcattcgtaa tgtctgccca ttctgctatt ctgtatacac
3421 ccgcagagta ctgcaatttg actgtattac caatgtcagc aaattttctg tcttcgaaga
3481 gtaaaaaatt gtacttggcg gataatgcct ttagcggctt aactgtgccc tccatggaaa
3541 aatcagtcaa gatatccaca tgtgttttta gtaaacaaat tttgggacct aatgcttcaa
3601 ctaactccag taattccttg gtggtacgaa catccaatga agcacacaag tttgtttgct
3661 tttcgtgcat gatattaaat agcttggcag caacaggact aggatgagta gcagcacgtt
3721 ccttatatgt agctttcgac atgatttatc ttcgtttcct gcaggttttt gttctgtgca
3781 gttgggttaa gaatactggg caatttcatg tttcttcaac actacatatg cgtatatata
3841 ccaatctaag tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa
3901 aaaaatttca aggaaaccga aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa
3961 ggtggtatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
4021 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
4081 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
4141 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
4201 aataatggtt tcttagacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa
4261 tgatggaata atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat
4321 tttagaaagt aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa
4381 ggtttaaaaa atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc
4441 agattaaata gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg
4501 tgtgtggtct tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa
4561 gagcaagata aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa
4621 aacaaaaact attttttctt taatttcttt ttttactttc tatttttaat ttatatattt
4681 atattaaaaa atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg
4741 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
4801 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
4861 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
4921 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
4981 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
5041 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
5101 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
5161 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
5221 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
5281 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
5341 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
5401 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
5461 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
5521 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
5581 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
5641 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
5701 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
5761 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
5821 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
5881 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
5941 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
6001 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
6061 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
6121 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
6181 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
6241 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
6301 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
6361 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
6421 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
6481 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
6541 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
6601 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
6661 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
6721 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
6781 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
6841 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
6901 gcgcgc
//
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