| 质粒名称: | pPP085 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 158795 |
| 存储实验室: | Jennifer Doudna |
|
载体骨架基本信息 |
|
| 载体名称: | pRSF-Duet1 |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 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: | casPhi-2 |
| 物种来源: | -- |
| 插入大小: | -- |
| 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: | pPP085 was a gift from Jennifer Doudna (Addgene plasmid # 158795 ; http://n2t.net/addgene:158795 ; RRID:Addgene_158795) |
| 参考文献部分: | CRISPR-CasPhi from huge phages is a hypercompact genome editor. Pausch P, Al-Shayeb B, Bisom-Rapp E, Tsuchida CA, Li Z, Cress BF, Knott GJ, Jacobsen SE, Banfield JF, Doudna JA. Science. 2020 Jul 17;369(6501):333-337. doi: 10.1126/science.abb1400. 10.1126/science.abb1400 PubMed 32675376 |
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 pPP085 6024 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION pRSF-Duet1 derived plasmid containing C-terminally hexa-histidine
tagged Cas-Phi-2 in MCS1 for expression in E. coli and protein
purification.
ACCESSION .
VERSION .
KEYWORDS pPP085
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6024)
AUTHORS Pausch P, Al-Shayeb B, Bisom-Rapp E, Tsuchida CA, Li Z, Cress BF,
Knott GJ, Jacobsen SE, Banfield JF, Doudna JA
TITLE CRISPR-CasPhi from huge phages is a hypercompact genome editor.
JOURNAL Science. 2020 Jul 17;369(6501):333-337. doi:
10.1126/science.abb1400.
PUBMED 32675376
REFERENCE 2 (bases 1 to 6024)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6024
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 549..568
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(588..665)
/gene="lacI"
/label=lacI promoter
promoter 711..729
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 730..754
/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)."
CDS 3069..3086
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
promoter 3136..3154
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 3155..3179
/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)."
CDS 3288..3332
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/label=S-Tag
/translation="KETAAAKFERQHMDS"
primer_bind complement(3370..3388)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 3384..3431
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(3661..4476)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 4384..4403
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
promoter complement(4477..4568)
/gene="bla"
/label=AmpR promoter
rep_origin complement(4584..5333)
/direction=LEFT
/label=RSF ori
/note="Plasmids containing the RSF 1030 origin of
replication can be propagated in E. coli cells that contain
additional plasmids with compatible origins."
primer_bind complement(4594..4611)
/label=L4440
/note="L4440 vector, forward primer"
primer_bind complement(4834..4853)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5529..587)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
ORIGIN
1 agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat
61 gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg
121 atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc
181 acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc
241 gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac
301 accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac
361 ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc
421 agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt
481 tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa
541 gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg
601 aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg
661 gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaaat taatacgact
721 cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt tgtttaactt
781 taataaggag atataccatg ccaaaacccg cggttgagag tgaattctcg aaagtgctta
841 agaagcactt ccccggggag cgcttccgtt ccagttacat gaaacgcggt ggcaagatct
901 tagccgccca gggagaagag gccgtagttg catacttaca gggaaaatca gaggaggaac
961 cgcctaactt ccaacctccc gcgaaatgtc atgtggttac taaaagtcgc gattttgccg
1021 aatggccgat tatgaaagcc agcgaggcta ttcaacgcta catctacgcg ttgtcaacca
1081 ctgagcgtgc cgcgtgtaaa ccagggaaaa gcagcgaatc acatgctgcc tggtttgcag
1141 cgactggcgt cagtaaccac gggtattcac atgtccaagg gcttaactta atctttgacc
1201 atactctggg gcgttacgat ggggttttga agaaggttca actgcgtaat gaaaaggcac
1261 gtgcccgttt ggagtcaatt aatgcatctc gtgccgatga gggtttacct gaaatcaagg
1321 cggaagagga agaggtagcc acaaatgaga cagggcattt gcttcaaccg ccaggcatta
1381 atcccagctt ctatgtttat caaacgatct cacctcaggc ttatcgtcca cgcgacgaga
1441 ttgttttgcc ccccgagtat gcagggtacg tgcgtgatcc caacgccccc attccgttag
1501 gagtagtccg caaccgttgt gatattcaga aaggctgtcc gggctatatt cctgagtggc
1561 agcgcgaggc aggtactgcg atctctccga agactgggaa agccgttaca gttcctggcc
1621 tttcccctaa gaagaacaag cgtatgcgcc gctattggcg ttcggaaaaa gagaaggcac
1681 aagatgcact gttggttacc gtccgtatcg gtactgactg ggtagtgatt gatgtgcgcg
1741 gattattgcg taatgcccgc tggcgcacca ttgctccaaa ggacatctca ttaaacgcgc
1801 ttttagacct gttcactgga gacccagtga ttgatgtccg ccgcaacatt gtaactttta
1861 cctacacgct tgatgcctgc ggtacctatg ctcgtaaatg gacgttaaaa ggaaagcaga
1921 caaaggcaac actggataaa ctgactgcca cccaaacagt agcgctggta gcgatcgatt
1981 tgggccagac gaaccctatt tcagcgggaa tctcccgtgt tacgcaggaa aacggggcgc
2041 ttcaatgtga gccgctggac cgctttacgt tacctgacga cttgttgaaa gatatcagtg
2101 cgtaccgcat cgcatgggac cgcaacgagg aggaattacg tgcccgttca gtcgaagcgt
2161 tacccgaagc acagcaggcc gaagtacgtg ctctggacgg cgtaagcaag gagaccgctc
2221 gtactcaatt atgtgctgat ttcggccttg atcccaagcg tctgccttgg gacaagatgt
2281 cctcaaacac tacattcatc agcgaagccc tgttgtcaaa tagtgtatcg cgtgaccagg
2341 ttttctttac cccagctcct aaaaaaggag ccaaaaaaaa agctccggtg gaggttatgc
2401 gcaaggaccg tacttgggcc cgtgcttata aaccacgtct gtctgttgaa gcccagaaac
2461 tgaaaaacga agctctgtgg gcccttaaac gtacgagtcc cgaatacctt aaattaagcc
2521 gtcgtaaaga ggagctttgc cgtcgcagta tcaattacgt tatcgagaag acccgccgtc
2581 gtacccagtg tcaaatcgtg atcccggtga ttgaagatct gaacgtccgc tttttccacg
2641 gatctgggaa acgtctgcct ggctgggata atttctttac agcgaaaaaa gagaaccgtt
2701 ggttcatcca gggccttcac aaggcttttt ccgacttacg cacacatcgc agtttctacg
2761 ttttcgaggt acgtccagag cgcacctcca ttacctgccc caaatgcggg cactgcgagg
2821 ttgggaaccg tgatggcgaa gcgttccaat gcctttcctg tggaaaaaca tgtaatgcag
2881 acttagacgt agccactcac aacctgacac aggttgccct taccggaaaa accatgccta
2941 aacgtgaaga accacgcgat gcccagggga ccgctccggc acgtaagacc aaaaaagcat
3001 caaagtcgaa ggccccccct gctgaacgcg aggaccagac cccggcccaa gaacctagcc
3061 agaccagcca tcaccatcac catcactaag tcgaacagaa agtaatcgta ttgtacacgg
3121 ccgcataatc gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc
3181 catcttagta tattagttaa gtataagaag gagatataca tatggcagat ctcaattgga
3241 tatcggccgg ccacgcgatc gctgacgtcg gtaccctcga gtctggtaaa gaaaccgctg
3301 ctgcgaaatt tgaacgccag cacatggact cgtctactag cgcagcttaa ttaacctagg
3361 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
3421 ggggtttttt gctgaaacct caggcatttg agaagcacac ggtcacactg cttccggtag
3481 tcaataaacc ggtaaaccag caatagacat aagcggctat ttaacgaccc tgccctgaac
3541 cgacgacaag ctgacgaccg ggtctccgca agtggcactt ttcggggaaa tgtgcgcgga
3601 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc
3661 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
3721 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
3781 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
3841 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
3901 tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca
3961 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
4021 cgcctgagcg agacgaaata cgcggtcgct gttaaaagga caattacaaa caggaatcga
4081 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
4141 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc
4201 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
4261 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
4321 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
4381 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
4441 cctagagcaa gacgtttccc gttgaatatg gctcatactc ttcctttttc aatattattg
4501 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
4561 taaacaaata ggcatgcagc gctcttccgc ttcctcgctc actgactcgc tacgctcggt
4621 cgttcgactg cggcgagcgg tgtcagctca ctcaaaagcg gtaatacggt tatccacaga
4681 atcaggggat aaagccggaa agaacatgtg agcaaaaagc aaagcaccgg aagaagccaa
4741 cgccgcaggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
4801 caagccagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
4861 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
4921 tcccttcggg aagcgtggcg ctttctcata gctcacgctg ttggtatctc agttcggtgt
4981 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
5041 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
5101 cagcagccat tggtaactga tttagaggac tttgtcttga agttatgcac ctgttaaggc
5161 taaactgaaa gaacagattt tggtgagtgc ggtcctccaa cccacttacc ttggttcaaa
5221 gagttggtag ctcagcgaac cttgagaaaa ccaccgttgg tagcggtggt ttttctttat
5281 ttatgagatg atgaatcaat cggtctatca agtcaacgaa cagctattcc gttactctag
5341 atttcagtgc aatttatctc ttcaaatgta gcacctgaag tcagccccat acgatataag
5401 ttgtaattct catgttagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
5461 ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg
5521 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
5581 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc
5641 accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc
5701 aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc
5761 gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat gtccgcacca
5821 acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca
5881 accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg
5941 gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga
6001 tatttatgcc agccagccag acgc
//
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