| 质粒名称: | p426-SNR52p-gRNA.csr-1.Y-SUP4t |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 68060 |
| 存储实验室: | Christian Hong |
|
载体骨架基本信息 |
|
| 载体名称: | pRS426 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 6274 bp |
| 载体修饰: | -- |
| 载体类型: | 酵母表达，CRISPR；N.crassa，真菌 |
| 筛选标记: | URA3 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | gRNA for csr-1 locus,gRNA,csr-1 |
| 物种来源: | N. crassa, S.pyogenes, other fungi |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | SNR52 |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | GGAGCTTCTTTGAAAAGATAATGTATG |
| 3' 测序引物 | GTTACATGCGTACACGCGTC |
|
文献引用方法 |
|
| 材料和方法部分: | p426-SNR52p-gRNA.csr-1.Y-SUP4t was a gift from Christian Hong (Addgene plasmid # 68060 ; http://n2t.net/addgene:68060 ; RRID:Addgene_68060) |
| 参考文献部分: | Efficient gene editing in Neurospora crassa with CRISPR technology. Matsu-ura Toru, Baek Mokryun, Kwon Jungin and Hong Christian. Fungal Biology and Biotechnology 2015, 2:4 10.1186/s40694-015-0015-1 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS p426-SNR52p-gRNA.csr-1.Y-SUP4t 6274 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION gRNA for csr-1 locus in N. crassa.
ACCESSION .
VERSION .
KEYWORDS p426-SNR52p-gRNA.csr-1.Y-SUP4t
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6274)
AUTHORS Matsu-ura Toru, Baek Mokryun, Kwon Jungin, Hong Christian
TITLE Efficient gene editing in Neurospora crassa with CRISPR technology
JOURNAL Fungal Biology and Biotechnology 2015, 2:4
REFERENCE 2 (bases 1 to 6274)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6274
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(10..28)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
rep_origin 69..1411
/label=2u ori
/note="yeast 2u plasmid origin of replication"
protein_bind 401..448
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
promoter 1438..1542
/gene="bla"
/label=AmpR promoter
CDS 1543..2403
/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(1761..1780)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2574..3162
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3063..3082
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3316..3333
/label=L4440
/note="L4440 vector, forward primer"
promoter 3486..3516
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3524..3540
/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 3529..3551
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 3548..3564
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3548..3564
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 3583..3603
/label=T3
/note="T3 promoter, forward primer"
promoter 3585..3603
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 3621..3889
/label=SNR52 promoter
/note="promoter for the S. cerevisiae small nucleolar RNA
gene SNR52"
misc_RNA 3910..3984
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
terminator 3989..4008
/label=SUP4 terminator
/note="transcription terminator for the S. cerevisiae SUP4
tRNA gene"
terminator 4014..4261
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind complement(4279..4298)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(4280..4298)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4308..4325)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4308..4324)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4317..4339)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 4469..4924
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4556..4575)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4766..4787
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(5055..5858)
/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(5859..6074)
/gene="S. cerevisiae URA3"
/label=URA3 promoter
primer_bind complement(6105..6124)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6224..6246
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
ORIGIN
1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
61 cttagtatga tccaatatca aaggaaatga tagcattgaa ggatgagact aatccaattg
121 aggagtggca gcatatagaa cagctaaagg gtagtgctga aggaagcata cgataccccg
181 catggaatgg gataatatca caggaggtac tagactacct ttcatcctac ataaatagac
241 gcatataagt acgcatttaa gcataaacac gcactatgcc gttcttctca tgtatatata
301 tatacaggca acacgcagat ataggtgcga cgtgaacagt gagctgtatg tgcgcagctc
361 gcgttgcatt ttcggaagcg ctcgttttcg gaaacgcttt gaagttccta ttccgaagtt
421 cctattctct agaaagtata ggaacttcag agcgcttttg aaaaccaaaa gcgctctgaa
481 gacgcacttt caaaaaacca aaaacgcacc ggactgtaac gagctactaa aatattgcga
541 ataccgcttc cacaaacatt gctcaaaagt atctctttgc tatatatctc tgtgctatat
601 ccctatataa cctacccatc cacctttcgc tccttgaact tgcatctaaa ctcgacctct
661 acatttttta tgtttatctc tagtattact ctttagacaa aaaaattgta gtaagaacta
721 ttcatagagt gaatcgaaaa caatacgaaa atgtaaacat ttcctatacg tagtatatag
781 agacaaaata gaagaaaccg ttcataattt tctgaccaat gaagaatcat caacgctatc
841 actttctgtt cacaaagtat gcgcaatcca catcggtata gaatataatc ggggatgcct
901 ttatcttgaa aaaatgcacc cgcagcttcg ctagtaatca gtaaacgcgg gaagtggagt
961 caggcttttt ttatggaaga gaaaatagac accaaagtag ccttcttcta accttaacgg
1021 acctacagtg caaaaagtta tcaagagact gcattataga gcgcacaaag gagaaaaaaa
1081 gtaatctaag atgctttgtt agaaaaatag cgctctcggg atgcattttt gtagaacaaa
1141 aaagaagtat agattctttg ttggtaaaat agcgctctcg cgttgcattt ctgttctgta
1201 aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca tttttgtttt
1261 acaaaaatga agcacagatt cttcgttggt aaaatagcgc tttcgcgttg catttctgtt
1321 ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag cgctctcgcg ttgcattttt
1381 gttctacaaa atgaagcaca gatgcttcgt tcaggtggca cttttcgggg aaatgtgcgc
1441 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
1501 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
1561 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
1621 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
1681 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
1741 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
1801 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
1861 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
1921 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
1981 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
2041 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
2101 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
2161 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
2221 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
2281 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
2341 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
2401 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
2461 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
2521 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
2581 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
2641 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
2701 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
2761 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
2821 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
2881 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
2941 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
3001 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
3061 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
3121 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
3181 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
3241 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
3301 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
3361 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
3421 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt acctcactca ttaggcaccc
3481 caggctttac actttatgct tccggctcct atgttgtgtg gaattgtgag cggataacaa
3541 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
3601 agggaacaaa agctggagct tctttgaaaa gataatgtat gattatgctt tcactcatat
3661 ttatacagaa acttgatgtt ttctttcgag tatatacaag gtgattacat gtacgtttga
3721 agtacaactc tagattttgt agtgccctct tgggctagcg gtaaaggtgc gcattttttc
3781 acaccctaca atgttctgtt caaaagattt tggtcaaacg ctgtagaagt gaaagttggt
3841 gcgcatgttt cggcgttcga aacttctccg cagtgaaaga taaatgatcg agtgggaggg
3901 tcccgtcctg ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
3961 tgaaaaagtg gcaccgagtc ggtggtgctt tttttgtttt ttatgtcttc gagtcatgta
4021 attagttatg tcacgcttac gttcacgccc tccccccaca tccgctctaa ccgaaaagga
4081 aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt
4141 aagaacgtta tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg
4201 taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg
4261 cggccggtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt
4321 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
4381 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
4441 ttgcgcagcc tgaatggcga atggcgcgac gcgccctgta gcggcgcatt aagcgcggcg
4501 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
4561 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
4621 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
4681 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
4741 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
4801 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
4861 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
4921 atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatagggt
4981 aataactgat ataattaaat tgaagctcta atttgtgagt ttagtataca tgcatttact
5041 tataatacag ttttttagtt ttgctggccg catcttctca aatatgcttc ccagcctgct
5101 tttctgtaac gttcaccctc taccttagca tcccttccct ttgcaaatag tcctcttcca
5161 acaataataa tgtcagatcc tgtagagacc acatcatcca cggttctata ctgttgaccc
5221 aatgcgtctc ccttgtcatc taaacccaca ccgggtgtca taatcaacca atcgtaacct
5281 tcatctcttc cacccatgtc tctttgagca ataaagccga taacaaaatc tttgtcgctc
5341 ttcgcaatgt caacagtacc cttagtatat tctccagtag atagggagcc cttgcatgac
5401 aattctgcta acatcaaaag gcctctaggt tcctttgtta cttcttctgc cgcctgcttc
5461 aaaccgctaa caatacctgg gcccaccaca ccgtgtgcat tcgtaatgtc tgcccattct
5521 gctattctgt atacacccgc agagtactgc aatttgactg tattaccaat gtcagcaaat
5581 tttctgtctt cgaagagtaa aaaattgtac ttggcggata atgcctttag cggcttaact
5641 gtgccctcca tggaaaaatc agtcaagata tccacatgtg tttttagtaa acaaattttg
5701 ggacctaatg cttcaactaa ctccagtaat tccttggtgg tacgaacatc caatgaagca
5761 cacaagtttg tttgcttttc gtgcatgata ttaaatagct tggcagcaac aggactagga
5821 tgagtagcag cacgttcctt atatgtagct ttcgacatga tttatcttcg tttcctgcag
5881 gtttttgttc tgtgcagttg ggttaagaat actgggcaat ttcatgtttc ttcaacacta
5941 catatgcgta tatataccaa tctaagtctg tgctccttcc ttcgttcttc cttctgttcg
6001 gagattaccg aatcaaaaaa atttcaagga aaccgaaatc aaaaaaaaga ataaaaaaaa
6061 aatgatgaat tgaattgaaa agctgtggta tggtgcactc tcagtacaat ctgctctgat
6121 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
6181 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
6241 cagaggtttt caccgtcatc accgaaacgc gcga
//
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