| 质粒名称: | pROS13 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 107927 |
| 存储实验室: | Jean-Marc Daran |
|
载体骨架基本信息 |
|
| 载体名称: | p426-SNR52p-gRNA.CAN1.Y-SUP4t |
| 载体出品公司: | George Church (Addgene plasmid # 43803) |
| 空载体大小: | 5817bp |
| 完整质粒大小: | 5809bp |
| 载体类型: | Yeast Expression, CRISPR |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | gRNA-CAN1.Y, gRNA-ADE2.Y |
| 物种来源: | S. cerevisiae (budding yeast) |
| 插入大小: | 20bp |
| 克隆方法: | Gibson Cloning |
|
文献引用方法 |
|
| 材料和方法部分: | pROS13 was a gift from Jean-Marc Daran (Addgene plasmid # 107927 ; http://n2t.net/addgene:107927 ; RRID:Addgene_107927) |
| 参考文献部分: | CRISPR/Cas9: a molecular Swiss army knife for simultaneous introduction of multiple genetic modifications in Saccharomyces cerevisiae. Mans R, van Rossum HM, Wijsman M, Backx A, Kuijpers NG, van den Broek M, Daran-Lapujade P, Pronk JT, van Maris AJ, Daran JM. FEMS Yeast Res. 2015 Mar;15(2). pii: fov004. doi: 10.1093/femsyr/fov004. Epub 2015 Mar 4. 10.1093/femsyr/fov004 PubMed 25743786 |
买家导航
pROS13载体质粒基本信息
pROS13质粒图谱载体图谱和pROS13载体序列质粒序列多克隆位点信息
pROS13质粒载体简介
pROS13质粒序列载体序列
LOCUS pROS13 5809 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION kan based Yeast/E.coli shuttle vector designed to clone and express
Cas9 programming spacer.
ACCESSION .
VERSION .
KEYWORDS pROS13
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5809)
AUTHORS Mans R, van Rossum HM, Wijsman M, Backx A, Kuijpers NG, van den
Broek M, Daran-Lapujade P, Pronk JT, van Maris AJ, Daran JM
TITLE CRISPR/Cas9: a molecular Swiss army knife for simultaneous
introduction of multiple genetic modifications in Saccharomyces
cerevisiae.
JOURNAL FEMS Yeast Res. 2015 Mar;15(2). pii: fov004. doi:
10.1093/femsyr/fov004. Epub 2015 Mar 4.
PUBMED 25743786
REFERENCE 2 (bases 1 to 5809)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5809
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(351..368)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(522..1110)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(602..621)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1281..2141)
/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 1904..1923
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2142..2246)
/gene="bla"
/label=AmpR promoter
protein_bind complement(2338..2371)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
gene 2426..3782
/label=kanMX
/note="yeast selectable marker conferring kanamycin
resistance (Wach et al., 1994)"
promoter 2426..2769
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
CDS 2770..3579
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(2837..2856)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
terminator 3585..3782
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
protein_bind complement(3845..3878)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
promoter 4001..4269
/label=SNR52 promoter
/note="promoter for the S. cerevisiae small nucleolar RNA
gene SNR52"
misc_RNA 4290..4364
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
terminator 4369..4388
/label=SUP4 terminator
/note="transcription terminator for the S. cerevisiae SUP4
tRNA gene"
rep_origin 4613..5594
/label=2u ori
/note="yeast 2u plasmid origin of replication"
terminator 5640..5659
/label=SUP4 terminator
/note="transcription terminator for the S. cerevisiae SUP4
tRNA gene"
misc_RNA complement(5664..5738)
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter complement(5759..218)
/label=SNR52 promoter
/note="promoter for the S. cerevisiae small nucleolar RNA
gene SNR52"
ORIGIN
1 caactttcac ttctacagcg tttgaccaaa atcttttgaa cagaacattg tagggtgtga
61 aaaaatgcgc acctttaccg ctagcccaag agggcactac aaaatctaga gttgtacttc
121 aaacgtacat gtaatcacct tgtatatact cgaaagaaaa catcaagttt ctgtataaat
181 atgagtgaaa gcataatcat acattatctt ttcaaagaag ctccagcttt tgttcccttt
241 agtgagggta ttcacgtaga cggataggta tagccagaca tcagcagcat acttcgggaa
301 ccgtaggctc tagagagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
361 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
421 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
481 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
541 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
601 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
661 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
721 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
781 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
841 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
901 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
961 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
1021 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
1081 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
1141 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
1201 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
1261 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
1321 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
1381 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
1441 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
1501 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
1561 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
1621 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
1681 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
1741 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
1801 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
1861 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
1921 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
1981 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
2041 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
2101 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
2161 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
2221 aaaaataaac aaataggggt tccgcgcact cgagactata tgtgaaggca tggctatggc
2281 acggcagaca ttccgccaga tcatcaatag gcacgcggcc ggtcgacaac ccttaatata
2341 acttcgtata atgtatgcta tacgaagtta ttaggtctag agatctgttt agcttgcctc
2401 gtccccgccg ggtcacccgg ccagcgacat ggaggcccag aataccctcc ttgacagtct
2461 tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg tacatttagc ccatacatcc
2521 ccatgtataa tcatttgcat ccatacattt tgatggccgc acggcgcgaa gcaaaaatta
2581 cggctcctcg ctgcagacct gcgagcaggg aaacgctccc ctcacagacg cgttgaattg
2641 tccccacgcc gcgcccctgt agagaaatat aaaaggttag gatttgccac tgaggttctt
2701 ctttcatata cttcctttta aaatcttgct aggatacagt tctcacatca catccgaaca
2761 taaacaacca tgggtaagga aaagactcac gtttcgaggc cgcgattaaa ttccaacatg
2821 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
2881 atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
2941 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
3001 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
3061 gcgatccccg gcaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
3121 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
3181 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
3241 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
3301 aaagaaatgc ataagctttt gccattctca ccggattcag tcgtcactca tggtgatttc
3361 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
3421 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
3481 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
3541 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagtactgac aataaaaaga
3601 ttcttgtttt caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt
3661 aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga
3721 agttaagtgc gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac
3781 tgctgtcgat tcgatactaa cgccgccatc cagtgtcgaa aacgagctct cgagaaccct
3841 taatataact tcgtataatg tatgctatac gaagttatta ggtgatatca gatccactag
3901 tggcctatgc cacctttcga gaggacgatg cccgtgtcta aatgattcga ccagcctaag
3961 aatgttcaac ccctcactaa agggaacaaa agctggagct tctttgaaaa gataatgtat
4021 gattatgctt tcactcatat ttatacagaa acttgatgtt ttctttcgag tatatacaag
4081 gtgattacat gtacgtttga agtacaactc tagattttgt agtgccctct tgggctagcg
4141 gtaaaggtgc gcattttttc acaccctaca atgttctgtt caaaagattt tggtcaaacg
4201 ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga aacttctccg cagtgaaaga
4261 taaatgatca cttgaagatt ctttagtgtg ttttagagct agaaatagca agttaaaata
4321 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtggtgctt tttttgtttt
4381 ttatgtcttc gagtcatgta attagttatg tcacgcttac gttcacgccc tccacgcatt
4441 taagcataaa cacgcactat gccgttcttc tcatgtatat atatatacag gcaacacgca
4501 gatataggtg cgacgtgaac agtgagctgt atgtgcgcag ctcgcgttgc attttcggaa
4561 gcgctcgttt tcggaaacgc tttgaagttc ctattccgaa gttcctattc tgtagaaagt
4621 ataggaacct cagagcgctt ttgaaaacca aaagcgctct gaagacgcac tttcaaaaaa
4681 ccaaaaacgc accggactgt aacgagctac taaaatattg cgaataccgc ttccacaaac
4741 attgctcaaa agtatctctt tgctatatat ctctgtgcta tatccctata taacctaccc
4801 atccaccttt cgctccttga acttgcatct aaactcgacc tctacatttt ttatgtttat
4861 ctctagtatt actctttaga caaaaaaatt gtagtaagaa ctattcatag agtgaatcga
4921 aaacaatacg aaaatgtaaa catttcctat acgtagtata tagagacaaa atagaagaaa
4981 ccgttcataa ttttctgacc aatgaagaat catcaacgct atcactttct gttcacaaag
5041 tatgcgcaat ccacatcggt atagaatata atcggggatg cctttatctt gaaaaaatgc
5101 acccgcagct tcgctagtaa tcagtaaacg cgggaagtgg agtcaggctt tttttatgga
5161 agagaaaata gacaccaaag tagccttctt ctaaccttaa cggacctaca gtgcaaaaag
5221 ttatcaagag actgcattat agagcgcaca aaggagaaaa aaagtaatct aagatgcttt
5281 gttagaaaaa tagcgctctc gggatgcatt tttgtagaac aaaaaagaag tatagattct
5341 ttgttggtaa aatagcgctc tcgcgttgca tttctgttct gtaaaaatgc agctcagatt
5401 ctttgtttga aaaattagcg ctctcgcgtt gcatttttgt tttacaaaaa tgaagcacag
5461 attcttcgtt ggtaaaatag cgctttcgcg ttgcatttct gttctgtaaa aatgcagctc
5521 agattctttg tttgaaaaat tagcgctctc gcgttgcatt tttgttctac aaaatgaagc
5581 acagatgctt cgttggaggg cgtgaacgta agcgtgacat aactaattac atgactcgaa
5641 gacataaaaa acaaaaaaag caccaccgac tcggtgccac tttttcaagt tgataacgga
5701 ctagccttat tttaacttgc tatttctagc tctaaaactc ctccatagag aacgtatcga
5761 tcatttatct ttcactgcgg agaagtttcg aacgccgaaa catgcgcac
//
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