| 质粒名称: | pROS16 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 107930 |
| 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: | 5462 bp |
| 完整质粒大小: | 5465 bp |
| 载体类型: | Yeast Expression, CRISPR |
| 筛选标记: | HIS3 |
| 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
gRNA/shRNA sequenceGATACGTTCTCTATGGAGGA, ACTTGAAGATTCTTTAGTGT
|
| 物种来源: | S. cerevisiae (budding yeast) |
| 插入大小: | 20 bp |
| 克隆方法: | Gibson Cloning |
|
文献引用方法 |
|
| 材料和方法部分: |
pROS16 was a gift from Jean-Marc Daran (Addgene plasmid # 107930 ; http://n2t.net/addgene:107930 ; RRID:Addgene_107930)
|
| 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
pROS16载体质粒基本信息
pROS16质粒图谱载体图谱和pROS16载体序列质粒序列多克隆位点信息
pROS16质粒载体简介
pROS16质粒序列载体序列
LOCUS pROS16 5465 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION HIS3 based Yeast/E.coli shuttle vector designed to clone and express
Cas9 programming spacer.
ACCESSION .
VERSION .
KEYWORDS pROS16
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5465)
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 5465)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5465
/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
primer_bind 2317..2336
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 2483..2669
/gene="S. cerevisiae HIS3"
/label=HIS3 promoter
CDS 2670..3332
/codon_start=1
/gene="S. cerevisiae HIS3"
/product="imidazoleglycerol-phosphate dehydratase, required
for histidine biosynthesis"
/label=HIS3
/note="yeast auxotrophic marker"
/translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
promoter 3657..3925
/label=SNR52 promoter
/note="promoter for the S. cerevisiae small nucleolar RNA
gene SNR52"
misc_RNA 3946..4020
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
terminator 4025..4044
/label=SUP4 terminator
/note="transcription terminator for the S. cerevisiae SUP4
tRNA gene"
rep_origin 4269..5250
/label=2u ori
/note="yeast 2u plasmid origin of replication"
terminator 5296..5315
/label=SUP4 terminator
/note="transcription terminator for the S. cerevisiae SUP4
tRNA gene"
misc_RNA complement(5320..5394)
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter complement(5415..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 gcactgcggc atcagagcag attgtactga
2341 gagtgcacca taaattcccg ttttaagagc ttggtgagcg ctaggagtca ctgccaggta
2401 tcgtttgaac acggcattag tcagggaagt cataacacag tcctttcccg caattttctt
2461 tttctattac tcttggcctc ctctagtaca ctctatattt ttttatgcct cggtaatgat
2521 tttcattttt tttttccacc tagcggatga ctcttttttt ttcttagcga ttggcattat
2581 cacataatga attatacatt atataaagta atgtgatttc ttcgaagaat atactaaaaa
2641 atgagcaggc aagataaacg aaggcaaaga tgacagagca gaaagcccta gtaaagcgta
2701 ttacaaatga aaccaagatt cagattgcga tctctttaaa gggtggtccc ctagcgatag
2761 agcactcgat cttcccagaa aaagaggcag aagcagtagc agaacaggcc acacaatcgc
2821 aagtgattaa cgtccacaca ggtatagggt ttctggacca tatgatacat gctctggcca
2881 agcattccgg ctggtcgcta atcgttgagt gcattggtga cttacacata gacgaccatc
2941 acaccactga agactgcggg attgctctcg gtcaagcttt taaagaggcc ctaggggccg
3001 tgcgtggagt aaaaaggttt ggatcaggat ttgcgccttt ggatgaggca ctttccagag
3061 cggtggtaga tctttcgaac aggccgtacg cagttgtcga acttggtttg caaagggaga
3121 aagtaggaga tctctcttgc gagatgatcc cgcattttct tgaaagcttt gcagaggcta
3181 gcagaattac cctccacgtt gattgtctgc gaggcaagaa tgatcatcac cgtagtgaga
3241 gtgcgttcaa ggctcttgcg gttgccataa gagaagccac ctcgcccaat ggtaccaacg
3301 atgttccctc caccaaaggt gttcttatgt agtgacaccg attatttaaa gctgcagcat
3361 acgatatata tacatgtgta tatatgtata cctatgaatg tcagtaagta tgtatacgaa
3421 cagtatgata ctgaagatga caaggtaatg catcattcta tacgtgtcat tctgaacgag
3481 gcgcgctttc cttttttctt tttgcttttt cttttttttt ctcttgaact cgacggatct
3541 atgcggtgtg aaataccgca cagatgcacc tttcgagagg acgatgcccg tgtctaaatg
3601 attcgaccag cctaagaatg ttcaacccct cactaaaggg aacaaaagct ggagcttctt
3661 tgaaaagata atgtatgatt atgctttcac tcatatttat acagaaactt gatgttttct
3721 ttcgagtata tacaaggtga ttacatgtac gtttgaagta caactctaga ttttgtagtg
3781 ccctcttggg ctagcggtaa aggtgcgcat tttttcacac cctacaatgt tctgttcaaa
3841 agattttggt caaacgctgt agaagtgaaa gttggtgcgc atgtttcggc gttcgaaact
3901 tctccgcagt gaaagataaa tgatcacttg aagattcttt agtgtgtttt agagctagaa
3961 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
4021 gtgctttttt tgttttttat gtcttcgagt catgtaatta gttatgtcac gcttacgttc
4081 acgccctcca cgcatttaag cataaacacg cactatgccg ttcttctcat gtatatatat
4141 atacaggcaa cacgcagata taggtgcgac gtgaacagtg agctgtatgt gcgcagctcg
4201 cgttgcattt tcggaagcgc tcgttttcgg aaacgctttg aagttcctat tccgaagttc
4261 ctattctgta gaaagtatag gaacctcaga gcgcttttga aaaccaaaag cgctctgaag
4321 acgcactttc aaaaaaccaa aaacgcaccg gactgtaacg agctactaaa atattgcgaa
4381 taccgcttcc acaaacattg ctcaaaagta tctctttgct atatatctct gtgctatatc
4441 cctatataac ctacccatcc acctttcgct ccttgaactt gcatctaaac tcgacctcta
4501 cattttttat gtttatctct agtattactc tttagacaaa aaaattgtag taagaactat
4561 tcatagagtg aatcgaaaac aatacgaaaa tgtaaacatt tcctatacgt agtatataga
4621 gacaaaatag aagaaaccgt tcataatttt ctgaccaatg aagaatcatc aacgctatca
4681 ctttctgttc acaaagtatg cgcaatccac atcggtatag aatataatcg gggatgcctt
4741 tatcttgaaa aaatgcaccc gcagcttcgc tagtaatcag taaacgcggg aagtggagtc
4801 aggctttttt tatggaagag aaaatagaca ccaaagtagc cttcttctaa ccttaacgga
4861 cctacagtgc aaaaagttat caagagactg cattatagag cgcacaaagg agaaaaaaag
4921 taatctaaga tgctttgtta gaaaaatagc gctctcggga tgcatttttg tagaacaaaa
4981 aagaagtata gattctttgt tggtaaaata gcgctctcgc gttgcatttc tgttctgtaa
5041 aaatgcagct cagattcttt gtttgaaaaa ttagcgctct cgcgttgcat ttttgtttta
5101 caaaaatgaa gcacagattc ttcgttggta aaatagcgct ttcgcgttgc atttctgttc
5161 tgtaaaaatg cagctcagat tctttgtttg aaaaattagc gctctcgcgt tgcatttttg
5221 ttctacaaaa tgaagcacag atgcttcgtt ggagggcgtg aacgtaagcg tgacataact
5281 aattacatga ctcgaagaca taaaaaacaa aaaaagcacc accgactcgg tgccactttt
5341 tcaagttgat aacggactag ccttatttta acttgctatt tctagctcta aaactcctcc
5401 atagagaacg tatcgatcat ttatctttca ctgcggagaa gtttcgaacg ccgaaacatg
5461 cgcac
//
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