| 质粒名称: | pBLO 64.1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 123120 |
| 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: | -- |
| 完整质粒大小: | 5559 bp |
| 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: | -- |
| 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 64.1 was a gift from Jennifer Doudna & Benjamin Oakes (Addgene plasmid # 123120 ; http://n2t.net/addgene:123120 ; RRID:Addgene_123120) |
| 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
pBLO64.1载体基本信息
pBLO64.1载体图谱质粒图谱和多克隆位点信息
pBLO64.1载体简介
pBLO64.1载体序列
LOCUS pBLO 64.1 5559 bp ds-DNA circular SYN 27-MAR-2019
DEFINITION E. coli expression vector for active deltaproteobacteria CasX.
ACCESSION .
VERSION .
KEYWORDS pBLO 64.1
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"
protein_bind 30..48
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
RBS 67..78
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
terminator 3069..3140
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 3156..3183
/label=T7Te terminator
/note="phage T7 early transcription terminator"
primer_bind complement(3211..3228)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3345..3890)
/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(3425..3444)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
terminator 4004..4098
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(4119..4778)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 4693..4712
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(4779..4881)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS complement(4911..5534)
/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 5442..5461
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
promoter 5553..49
/gene="tetR"
/label=tetR/tetA promoters
/note="overlapping promoters for bacterial tetR and tetA"
ORIGIN
1 ctctatcgtt gatagagtta ttttaccact ccctatcagt gatagagaaa agaattcaaa
61 agatctaaag aggagaaagg atctatggaa aagagaatca acaagatcag gaagaagctg
121 agcgccgaca acgccaccaa gcctgtgtct aggagtggcc ccatgaaaac cctgctcgtc
181 agagtgatga ccgacgacct gaagaagaga ctggaaaaga ggaggaagaa acccgaagtg
241 atgccccaag tgatcagcaa caacgccgcc aacaacctga gaatgctgct ggacgactac
301 accaagatga aggaagccat cctgcaggtc tactggcaag agttcaagga cgaccacgtg
361 ggcctgatgt gcaagtttgc ccagcctgcc agcaagaaga tcgaccagaa caagctgaag
421 cccgagatgg acgagaaggg caatctgacc acagccggct ttgcctgctc tcagtgtggc
481 cagcctctgt tcgtgtacaa gctggaacag gtgtccgaga aaggcaaggc ctacaccaac
541 tacttcggca gatgtaacgt ggccgagcac gagaagctga ttctgctggc ccagctgaaa
601 cctgagaagg actctgatga ggccgtgacc tacagcctgg gcaagtttgg acagagagcc
661 ctggacttct acagcatcca cgtgaccaaa gaaagcacac accccgtgaa gcccctggct
721 cagatcgccg gcaatagata cgcctctgga cctgtgggca aagccctgtc cgatgcctgc
781 atgggaacaa tcgccagctt cctgagcaag taccaggaca tcatcatcga gcaccagaag
841 gtggtcaagg gcaaccagaa gagactggaa agcctgaggg agctggccgg caaagagaac
901 ctggaatacc ccagcgtgac cctgcctcct cagcctcaca caaaagaagg cgtggacgcc
961 tacaacgaag tgatcgccag agtgagaatg tgggtcaacc tgaacctgtg gcagaagctg
1021 aaactgtcca gggacgacgc caagcctctg ctgagactga agggcttccc tagcttcccc
1081 gtggtggaaa gaagggagaa tgaggtggac tggtggaaca ccatcaacga agtcaagaaa
1141 ctgatcgatg ccaagagaga catgggcaga gtgttttgga gcggagtgac cgccgagaag
1201 agaaacacca tcctggaagg ctacaactac ctgcctaacg agaacgacca caagaagaga
1261 gagggcagcc tggaaaaccc taagaagccc gccaagagac agttcggcga cctgctgctg
1321 tacctggaga agaagtatgc cggcgactgg ggcaaagtgt tcgatgaggc ctgggagaga
1381 atcgacaaga agattgccgg cctgaccagc cacatcgaga gagaggaagc cagaaacgcc
1441 gaggacgccc agtctaaagc cgtgctgaca gattggctga gagccaaggc cagcttcgtg
1501 ctggaaaggc tgaaagaaat ggatgagaaa gagttctacg cctgcgagat ccagctgcag
1561 aagtggtacg gggacctgag aggcaatccc tttgccgtgg aagccgagaa cagagtggtg
1621 gacatcagcg gctttagcat cggctctgac ggccactcca tccagtacag gaacctgctg
1681 gcctggaagt acctggaaaa tggcaagaga gagttctacc tgctgatgaa ctacggcaag
1741 aagggcagaa tcagattcac cgacggcacc gacatcaaga aaagcggcaa gtggcagggc
1801 ctgctgtatg gcggcggaaa ggccaaagtg atcgacctga ccttcgatcc cgacgacgag
1861 cagctgatca ttctgcctct ggccttcggc accagacagg gcagagagtt catctggaac
1921 gatctgctgt ccctggaaac cggcctgatc aagctggcca acggcagggt gatcgagaaa
1981 accatctaca acaagaaaat cggcagagat gagcccgcac tgtttgtggc cctgaccttt
2041 gagaggagag aagtggtcga tcccagcaac atcaagcccg tgaacctgat cggagtggac
2101 agaggcgaga atatccctgc cgtgattgcc ctgacagacc ctgagggatg tcccctgcct
2161 gagtttaagg atagctctgg cggccctacc gacatcctga gaatcggcga gggctacaaa
2221 gagaagcaga gagccatcca ggccgccaaa gaggtggaac agagaagagc cggcggatac
2281 agcagaaagt tcgccagcaa gtccagaaac ctggccgatg acatggtcag gaacagcgcc
2341 agggacctgt tctaccacgc cgtgacacat gatgccgtgc tggtgttcga gaacctgtca
2401 agaggcttcg gcagacaggg aaaaagaacc ttcatgaccg agagacagta cacgaagatg
2461 gaagattggc tcaccgccaa actggcctac gagggcctga caagcaagac ctacctgtcc
2521 aagacactgg cccagtacac ctccaagacc tgctccaatt gcggcttcac catcaccacc
2581 gccgactacg atggaatgct cgtgagactg aagaaaacct ccgacggctg ggccaccaca
2641 ctgaacaaca aagagctgaa agccgagggc cagatcacct actacaacag atacaagagg
2701 cagaccgtcg agaaagagct gtccgccgaa ctcgacagac tgagcgagga atccggcaac
2761 aacgacatca gcaagtggac caagggacgc agagatgagg ccctgttcct gctgaagaag
2821 aggttcagcc acagacctgt gcaagagcag tttgtgtgcc tggattgcgg ccacgaagtg
2881 cacgcagatg aacaggccgc tctgaatatc gccagaagct ggctgttcct gaacagcaac
2941 agcaccgagt tcaagagcta caagagcggc aagcagcctt tcgttggagc ctggcaggcc
3001 ttctacaaga gaaggctcaa agaagtgtgg aagcccaacg cctgactcga gtaaggatct
3061 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
3121 gtttgtcggt gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct
3181 gcgtttatac ctagggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt
3241 tcgactgcgg cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag
3301 gaagatactt aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc
3361 ccccctgaca agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga
3421 ctataaagat accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc
3481 tttcggttta ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac
3541 actcagttcc gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca
3601 gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc
3661 aaaagcacca ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat
3721 gcgccggtta aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt
3781 tacctcggtt caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg
3841 ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc
3901 ttattaatca gataaaatat ttctagattt cagtgcaatt tatctcttca aatgtagcac
3961 ctgaagtcag ccccatacga tataagttgt tactagtgct tggattctca ccaataaaaa
4021 acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg
4081 gatctatcaa caggagtcca agcgagctcg atatcaaatt acgccccgcc ctgccactca
4141 tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca
4201 tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc
4261 atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg
4321 aaactcaccc agggattggc tgacacgaaa aacatattct caataaaccc tttagggaaa
4381 taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg
4441 aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg
4501 gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacga
4561 aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg
4621 tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta
4681 taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat
4741 atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa
4801 aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg
4861 gaacctctta cgtgccgatc aacgtttcat tttcgccaga tatcgacgtc ttaagaccca
4921 ctttcacatt taagttgttt ttctaatccg catatgatca attcaaggcc gaataagaag
4981 gctggctctg caccttggtg atcaaataat tcgatagctt gtcgtaataa tggcggcata
5041 ctatcagtag taggtgtttc cctttcttct ttagcgactt gatgctcttg atcttccaat
5101 acgcaaccta aagtaaaatg ccccacagcg ctgagtgcat ataatgcatt ctctagtgaa
5161 aaaccttgtt ggcataaaaa ggctaattga ttttcgagag tttcatactg tttttctgta
5221 ggccgtgtac ctaaatgtac ttttgctcca tcgcgatgac ttagtaaagc acatctaaaa
5281 cttttagcgt tattacgtaa aaaatcttgc cagctttccc cttctaaagg gcaaaagtga
5341 gtatggtgcc tatctaacat ctcaatggct aaggcgtcga gcaaagcccg cttatttttt
5401 acatgccaat acaatgtagg ctgctctaca cctagcttct gggcgagttt acgggttgtt
5461 aaaccttcga ttccgacctc attaagcagc tctaatgcgc tgttaatcac tttactttta
5521 tctaatctag acatcattaa ttcctaattt ttgttgaca
//
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