| 质粒名称: | pBLO ddd64.1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 123121 |
| 存储实验室: | Jennifer Doudna, Benjamin Oakes |
|
载体骨架基本信息 |
|
| 载体名称: | -- |
| 载体出品公司: | -- |
| 载体抗性: | 氯霉素 |
| 空载体大小: | 5559 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | CRISPR |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | CasX |
| 物种来源: | Deltaproteobacteria |
| 插入大小: | 2955 bp |
| 基因突变: | Mutated amino acids in catalytic triad to alanine at positions 672, 769, and 935 |
| GenBank编号 | -- |
| 启动子 | -- |
| 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 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pBLO ddd64.1 was a gift from Jennifer Doudna & Benjamin Oakes (Addgene plasmid # 123121 ; http://n2t.net/addgene:123121 ; RRID:Addgene_123121) |
| 参考文献部分: | CasX enzymes comprise a distinct family of RNA-guided genome editors. Liu JJ, Orlova N, Oakes BL, Ma E, Spinner HB, Baney KLM, Chuck J, Tan D, Knott GJ, Harrington LB, Al-Shayeb B, Wagner A, Brotzmann J, Staahl BT, Taylor KL, Desmarais J, Nogales E, Doudna JA. Nature. 2019 Feb 4. pii: 10.1038/s41586-019-0908-x. doi: 10.1038/s41586-019-0908-x. 10.1038/s41586-019-0908-x PubMed 30718774 |
买家导航
pBLOddd64.1载体基本信息
pBLOddd64.1载体图谱质粒图谱和多克隆位点信息
pBLOddd64.1载体简介
pBLOddd64.1载体序列
LOCUS pBLO ddd64.1 5559 bp ds-DNA circular SYN 27-MAR-2019
DEFINITION E. coli expression vector for catalytically inactivated
deltaproteobacteria CasX.
ACCESSION .
VERSION .
KEYWORDS pBLO ddd64.1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5559)
AUTHORS Liu JJ, Orlova N, Oakes BL, Ma E, Spinner HB, Baney KLM, Chuck J,
Tan D, Knott GJ, Harrington LB, Al-Shayeb B, Wagner A, Brotzmann J,
Staahl BT, Taylor KL, Desmarais J, Nogales E, Doudna JA
TITLE CasX enzymes comprise a distinct family of RNA-guided genome
editors.
JOURNAL Nature. 2019 Feb 4. pii: 10.1038/s41586-019-0908-x. doi:
10.1038/s41586-019-0908-x.
PUBMED 30718774
REFERENCE 2 (bases 1 to 5559)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Mar 27, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5559
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 2668..2739
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2755..2782
/label=T7Te terminator
/note="phage T7 early transcription terminator"
primer_bind complement(2810..2827)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2944..3489)
/direction=LEFT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin.
"
primer_bind complement(3024..3043)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
terminator 3603..3697
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(3718..4377)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 4292..4311
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(4378..4480)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS complement(4510..5133)
/codon_start=1
/gene="tetR from transposon Tn10"
/product="tetracycline repressor TetR"
/label=TetR
/note="TetR binds to the tetracycline operator tetO to
inhibit transcription. This inhibition can be relieved by
adding tetracycline or doxycycline."
/translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
primer_bind 5041..5060
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
promoter 5152..5207
/gene="tetR"
/label=tetR/tetA promoters
/note="overlapping promoters for bacterial tetR and tetA"
protein_bind 5188..5206
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
RBS 5225..5236
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
ORIGIN
1 cgaccagaac aagctgaagc ccgagatgga cgagaagggc aatctgacca cagccggctt
61 tgcctgctct cagtgtggcc agcctctgtt cgtgtacaag ctggaacagg tgtccgagaa
121 aggcaaggcc tacaccaact acttcggcag atgtaacgtg gccgagcacg agaagctgat
181 tctgctggcc cagctgaaac ctgagaagga ctctgatgag gccgtgacct acagcctggg
241 caagtttgga cagagagccc tggacttcta cagcatccac gtgaccaaag aaagcacaca
301 ccccgtgaag cccctggctc agatcgccgg caatagatac gcctctggac ctgtgggcaa
361 agccctgtcc gatgcctgca tgggaacaat cgccagcttc ctgagcaagt accaggacat
421 catcatcgag caccagaagg tggtcaaggg caaccagaag agactggaaa gcctgaggga
481 gctggccggc aaagagaacc tggaataccc cagcgtgacc ctgcctcctc agcctcacac
541 aaaagaaggc gtggacgcct acaacgaagt gatcgccaga gtgagaatgt gggtcaacct
601 gaacctgtgg cagaagctga aactgtccag ggacgacgcc aagcctctgc tgagactgaa
661 gggcttccct agcttccccg tggtggaaag aagggagaat gaggtggact ggtggaacac
721 catcaacgaa gtcaagaaac tgatcgatgc caagagagac atgggcagag tgttttggag
781 cggagtgacc gccgagaaga gaaacaccat cctggaaggc tacaactacc tgcctaacga
841 gaacgaccac aagaagagag agggcagcct ggaaaaccct aagaagcccg ccaagagaca
901 gttcggcgac ctgctgctgt acctggagaa gaagtatgcc ggcgactggg gcaaagtgtt
961 cgatgaggcc tgggagagaa tcgacaagaa gattgccggc ctgaccagcc acatcgagag
1021 agaggaagcc agaaacgccg aggacgccca gtctaaagcc gtgctgacag attggctgag
1081 agccaaggcc agcttcgtgc tggaaaggct gaaagaaatg gatgagaaag agttctacgc
1141 ctgcgagatc cagctgcaga agtggtacgg ggacctgaga ggcaatccct ttgccgtgga
1201 agccgagaac agagtggtgg acatcagcgg ctttagcatc ggctctgacg gccactccat
1261 ccagtacagg aacctgctgg cctggaagta cctggaaaat ggcaagagag agttctacct
1321 gctgatgaac tacggcaaga agggcagaat cagattcacc gacggcaccg acatcaagaa
1381 aagcggcaag tggcagggcc tgctgtatgg cggcggaaag gccaaagtga tcgacctgac
1441 cttcgatccc gacgacgagc agctgatcat tctgcctctg gccttcggca ccagacaggg
1501 cagagagttc atctggaacg atctgctgtc cctggaaacc ggcctgatca agctggccaa
1561 cggcagggtg atcgagaaaa ccatctacaa caagaaaatc ggcagagatg agcccgcact
1621 gtttgtggcc ctgacctttg agaggagaga agtggtcgat cccagcaaca tcaagcccgt
1681 gaacctgatc ggagtggcca gaggcgagaa tatccctgcc gtgattgccc tgacagaccc
1741 tgagggatgt cccctgcctg agtttaagga tagctctggc ggccctaccg acatcctgag
1801 aatcggcgag ggctacaaag agaagcagag agccatccag gccgccaaag aggtggaaca
1861 gagaagagcc ggcggataca gcagaaagtt cgccagcaag tccagaaacc tggccgatga
1921 catggtcagg aacagcgcca gggacctgtt ctaccacgcc gtgacacatg atgccgtgct
1981 ggtgttcgcg aacctgtcaa gaggcttcgg cagacaggga aaaagaacct tcatgaccga
2041 gagacagtac acgaagatgg aagattggct caccgccaaa ctggcctacg agggcctgac
2101 aagcaagacc tacctgtcca agacactggc ccagtacacc tccaagacct gctccaattg
2161 cggcttcacc atcaccaccg ccgactacga tggaatgctc gtgagactga agaaaacctc
2221 cgacggctgg gccaccacac tgaacaacaa agagctgaaa gccgagggcc agatcaccta
2281 ctacaacaga tacaagaggc agaccgtcga gaaagagctg tccgccgaac tcgacagact
2341 gagcgaggaa tccggcaaca acgacatcag caagtggacc aagggacgca gagatgaggc
2401 cctgttcctg ctgaagaaga ggttcagcca cagacctgtg caagagcagt ttgtgtgcct
2461 ggattgcggc cacgaagtgc acgcagctga acaggccgct ctgaatatcg ccagaagctg
2521 gctgttcctg aacagcaaca gcaccgagtt caagagctac aagagcggca agcagccttt
2581 cgttggagcc tggcaggcct tctacaagag aaggctcaaa gaagtgtgga agcccaacgc
2641 ctgactcgag taaggatctc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
2701 gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca
2761 ccttcgggtg ggcctttctg cgtttatacc tagggatata ttccgcttcc tcgctcactg
2821 actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa cggggcggag
2881 atttcctgga agatgccagg aagatactta acagggaagt gagagggccg cggcaaagcc
2941 gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct caaatcagtg
3001 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggcg gctccctcgt
3061 gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt
3121 gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag ctggactgta
3181 tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat cgtcttgagt
3241 ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat tgatttagag
3301 gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag ttttggtgac
3361 tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag aaccttcgaa
3421 aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg cagaccaaaa
3481 cgatctcaag aagatcatct tattaatcag ataaaatatt tctagatttc agtgcaattt
3541 atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgtt actagtgctt
3601 ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg
3661 agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctcga tatcaaatta
3721 cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg
3781 gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc
3841 ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac
3901 gtttaaatca aaactggtga aactcaccca gggattggct gacacgaaaa acatattctc
3961 aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata
4021 tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc
4081 agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc
4141 gtctttcatt gccatacgaa attccggatg agcattcatc aggcgggcaa gaatgtgaat
4201 aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc
4261 cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc
4321 tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt
4381 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat
4441 ttcattatgg tgaaagttgg aacctcttac gtgccgatca acgtttcatt ttcgccagat
4501 atcgacgtct taagacccac tttcacattt aagttgtttt tctaatccgc atatgatcaa
4561 ttcaaggccg aataagaagg ctggctctgc accttggtga tcaaataatt cgatagcttg
4621 tcgtaataat ggcggcatac tatcagtagt aggtgtttcc ctttcttctt tagcgacttg
4681 atgctcttga tcttccaata cgcaacctaa agtaaaatgc cccacagcgc tgagtgcata
4741 taatgcattc tctagtgaaa aaccttgttg gcataaaaag gctaattgat tttcgagagt
4801 ttcatactgt ttttctgtag gccgtgtacc taaatgtact tttgctccat cgcgatgact
4861 tagtaaagca catctaaaac ttttagcgtt attacgtaaa aaatcttgcc agctttcccc
4921 ttctaaaggg caaaagtgag tatggtgcct atctaacatc tcaatggcta aggcgtcgag
4981 caaagcccgc ttatttttta catgccaata caatgtaggc tgctctacac ctagcttctg
5041 ggcgagttta cgggttgtta aaccttcgat tccgacctca ttaagcagct ctaatgcgct
5101 gttaatcact ttacttttat ctaatctaga catcattaat tcctaatttt tgttgacact
5161 ctatcgttga tagagttatt ttaccactcc ctatcagtga tagagaaaag aattcaaaag
5221 atctaaagag gagaaaggat ctatggaaaa gagaatcaac aagatcagga agaagctgag
5281 cgccgacaac gccaccaagc ctgtgtctag gagtggcccc atgaaaaccc tgctcgtcag
5341 agtgatgacc gacgacctga agaagagact ggaaaagagg aggaagaaac ccgaagtgat
5401 gccccaagtg atcagcaaca acgccgccaa caacctgaga atgctgctgg acgactacac
5461 caagatgaag gaagccatcc tgcaggtcta ctggcaagag ttcaaggacg accacgtggg
5521 cctgatgtgc aagtttgccc agcctgccag caagaagat
//
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