| 质粒名称: | PL6-CS-WR |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 122512 |
| 存储实验室: | Lubin Jiang |
|
载体骨架基本信息 |
|
| 载体名称: | PL6 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6676 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | CRISPR |
| 筛选标记: | hDHFR |
| 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: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | PfU6 |
| 标签/融合蛋白: | -- |
| 克隆方法: | Ligation Independent Cloning |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | PL6-CS-WR was a gift from Lubin Jiang (Addgene plasmid # 122512 ; http://n2t.net/addgene:122512 ; RRID:Addgene_122512) |
| 参考文献部分: | Epigenetic editing by CRISPR/dCas9 in Plasmodium falciparum. Xiao B, Yin S, Hu Y, Sun M, Wei J, Huang Z, Wen Y, Dai X, Chen H, Mu J, Cui L, Jiang L. Proc Natl Acad Sci U S A. 2019 Jan 2;116(1):255-260. doi: 10.1073/pnas.1813542116. Epub 2018 Dec 24. 10.1073/pnas.1813542116 PubMed 30584102 |
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 PL6-CS-WR 6676 bp ds-DNA circular SYN 24-JAN-2019
DEFINITION construction for sgRNA transcription with Plasmodium falciparum U6
promotor.
ACCESSION .
VERSION .
KEYWORDS PL6-CS-WR
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6676)
AUTHORS Xiao B, Yin S, Hu Y, Sun M, Wei J, Huang Z, Wen Y, Dai X, Chen H, Mu
J, Cui L, Jiang L
TITLE Epigenetic editing by CRISPR/dCas9 in Plasmodium falciparum.
JOURNAL Proc Natl Acad Sci U S A. 2019 Jan 2;116(1):255-260. doi:
10.1073/pnas.1813542116. Epub 2018 Dec 24.
PUBMED 30584102
REFERENCE 2 (bases 1 to 6676)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jan 24, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6676
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 2364..2385
/label=EGFP-C
/note="EGFP, forward primer"
misc_RNA 3621..3696
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
primer_bind complement(4223..4242)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4342..4364
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4402..4420)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4488..4592
/gene="bla"
/label=AmpR promoter
CDS 4593..5453
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(4811..4830)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5624..6212
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6113..6132
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6366..6383
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 6500..6521
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6536..6566
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6574..6590
/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 6579..6601
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 6598..6614
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 6598..6614
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 6632..6650
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 6632..6649
/label=SP6
/note="SP6 promoter, forward primer"
ORIGIN
1 ggcgcgccat gtaggcgcca actcttaagc attttgtaaa aaaaattaaa atatatttat
61 ataatattat tttattttat tatatattat attattttta tttttatttt tatttttttt
121 tctctacaaa ttttatctat tggtttatta taaaaatatc tatttctaat aataaataat
181 taagatatca atttatagaa acaaaatata tacttgtata attttatttt tttatataaa
241 tcattacata tataattata caatattttt tctaagagat aattatatat taatatatat
301 aaaaaaaggt gttttttttt ttttttttta tttttatttt tattttatgg taatatttta
361 ttttccttat tttataaatt atattagttt atatgtgatt aattttatat attatcaatt
421 tatatatttt taaatgctta cttaattatc tttttttttt tttttttttt tttttcccct
481 ctttttatat taatttattt ttgaaaaaat tgatatatat atatatatat aatatatata
541 tatacatgta gtagtattaa acaatgtata atatatataa ataatatatt tatatatttc
601 atttcaattt taattttttt tggttttttt tttttttctt tttgtcatat ttaaaaaaaa
661 ttatattcat ataagttatg cattttttat aaacattatt caatatatgt ataatataat
721 atatatatat atattaatgt attattccaa tgtgcatgat aaaagaaaaa aataatattt
781 ataaaaaaaa agaaaaataa aacaaaaaaa gaaaaaaaaa aaaaaaaaaa aaaaaataca
841 aaaataaata atataattta taattatata ttcttgtcac aataaaaata tatatatata
901 tatatatatt tataatatgt atattttaaa ctagaaaagg aataactaat attttattta
961 ttatcattca agatttatat tttataataa taaataccta atagaaatat atcaggatcc
1021 atgcatggtt cgctaaactg catcgtcgct gtgtcccaga acatgggcat cggcaagaac
1081 ggggactacc cctggccacc gctcaggaac gaatttagat atttccagag aatgaccaca
1141 acctcttcag tagaaggtaa acagaatctg gtgattatgg gtaagaagac ctggttctcc
1201 attcctgaga agaatcgacc tttaaagggt agaattaatt tagttctcag cagagaactc
1261 aaggaacctc cacaaggagc tcattttctt tccagaagtc tagatgatgc cttaaaactt
1321 actgaacaac cagaattagc aaataaagta gacatggtct ggatagttgg tggcagttct
1381 gtttataagg aagccatgaa tcacccaggc catcttaaac tatttgtgac aaggatcatg
1441 caagactttg aaagtgacac gttttttcca gaaattgatt tggagaaata taaacttctg
1501 ccagaatacc caggtgttct ctctgatgtc caggaggaga aaggcattaa gtacaaattt
1561 gaagtatatg agaagaatga ttaaaagctt atttaataat agattaaaaa tattataaaa
1621 ataaaaacat aaacacagaa attacaaaaa aaatacatat gaattttttt tttgtaatct
1681 tccttataaa tatagaataa tgaatcatat aaaacatatc attattcatt tatttacatt
1741 taaaattatt gtttcagtat ctttaattta ttatgtatat ataaaaataa cttacaattt
1801 tattaataaa caatatatgt ttattaattc atgttttgta atttatggga tagcgatttt
1861 ttttactgtc tgtatttttc ttttttaatt atgttttaat tgtattttat ttttattatt
1921 gttcttttta tagtattatt ttaaaacaaa atgtattttc taagaactta taataataat
1981 aaatataaat tttaataaaa attatattta tcttttacaa tatgaacata aagtacaaca
2041 ttaatatata gcttttaata tttttattcc taatcatgta aatcttaaat ttttcttttt
2101 aaacatatgt taaatattta tttctcatta tatataagaa catatttatt aaatctagaa
2161 ttctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
2221 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
2281 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
2341 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
2401 actctcggca tggacgagct gtacaagtaa ccatggaacg gtaaaaataa taacacgaat
2461 tatcacaata atttttcaca tatgtatata tatatatata ttatcaaagg gacaaccgtt
2521 tcaagaaagc aaatgttatt attttgtttt tattctaatg tatgtaagcc aaaaaccaat
2581 aaaaaagtaa taattaaaaa ggaaataaaa taaatataca tatgtaatta tacttatata
2641 ataagtatgt atatttttta ttattttttt tttttaatgc tatttattta ttattgcatc
2701 tttttattta atttttgtat ttttttatat aattccttta tttctttatt tctttcttta
2761 tttctttttt tttttttttt tttttttttt tttcctttaa ttaaacttta aaagacgtta
2821 taaattttaa acatatttta taaataaaaa aaaaaaaaaa aaagacgaaa taataaaata
2881 aacgaagata aagtaatatt ttgttattat gtcacatagt atgtgtacta caattattta
2941 tggatgcaaa ttattaaatt ggtagttatt atatacatac atgttttttt ttttgtgtgt
3001 attaaaagga actattatat aaatagaaca atttcatgac atttggattt ctacacatct
3061 tgaggtttta caataatatc ttcttttttt ttttttctct tctatatatg aaatatattg
3121 taaaaaggta caatatatat aatatttata tactatgaaa aaaaaaaaat atatacataa
3181 actaataata atttattcta atttaatata aaaaaaaatt cttgcttgtt cagataaata
3241 aaagaaattt atataaatag ctatattttt tttttttttt tttttttttt tttctaatta
3301 taaatataat tattcattta aaaaatttta aatatttaaa tatttcataa attaaaaaaa
3361 aaaaaaaaaa aactattaat ataataaact tttattttta ctgtaatata attttttata
3421 atgtaaaaat aaagggtaaa ttattattaa aaaatgtata tgttatgtat atataacata
3481 atatattata atatatatat atatatatat atatatataa tattagagta accaaaatgc
3541 ataatttttc ctatatgcac atatttcata ttaagtatat aatattccta ggaactcatc
3601 gctcgcgatg ctgcccgaca gttttagagc tagaaatagc aagttaaaat aaggctagtc
3661 cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttattatttc ctataaaata
3721 atatatatat atatatatat atatatatat atatatttta aatataatga agtttttaag
3781 gctaccataa aaaaaaaaat acatatatat ataaataaat atattatatt atataatata
3841 tatattaatg tttttgtttt ttaatatttt tttttttttt tgtaaaatat attatttttg
3901 gacttataaa taattgtgta catacatata ttattttatt ataataaata taataatact
3961 ataataatct ttatgcaata tttgtaattc atatataata atttttaaaa ttatctatct
4021 tttgaaaaaa aaaaaaaaat aaaaataata aaaattataa tgttattaaa tactataatt
4081 attatgatta atataatata tatatatata tatatatata tatatatata tatatatttt
4141 atatcattta atatattatg tgtgatataa taataaatcc attgtccata aatagatgaa
4201 atagcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
4261 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
4321 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
4381 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
4441 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
4501 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
4561 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
4621 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
4681 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
4741 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
4801 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
4861 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
4921 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
4981 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
5041 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
5101 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatgccaaca acgttgcgca
5161 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
5221 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
5281 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
5341 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
5401 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
5461 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
5521 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
5581 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
5641 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
5701 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
5761 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
5821 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
5881 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
5941 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
6001 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
6061 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
6121 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
6181 gatgctcgtc aggggggcgg agcctatcga aaaacgccag caacgcggcc tttttacggt
6241 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
6301 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
6361 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
6421 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
6481 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
6541 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
6601 gaaacagcta tgaccatgat tacgccaagc tatttaggtg acactataga atactcaagc
6661 tgcggcccta gtctag
//
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