| 质粒名称: | pAG/MNase |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 123461 |
| 存储实验室: | Steven Henikoff |
|
载体骨架基本信息 |
|
| 载体名称: | pK19pA-MN |
| 载体出品公司: | Ulrich Laemmli |
| 完整质粒大小: | 3846 bp |
| 载体类型: | 细菌细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | JM101 |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | Protein A-Protein G-Micrococcal Nuclease/pAG-MNase/pAG/MN/pAG(ERH)_MNase |
| 插入大小: | 1218 bp |
| 启动子: | Lac |
| 标签/融合蛋白: |
6xHIS (C terminal on insert)
HA (C terminal on insert)
|
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | M13R |
| 3’测序引物 | M13F |
|
文献引用方法 |
|
| 材料和方法部分: | pAG/MNase was a gift from Steven Henikoff (Addgene plasmid # 123461 ; http://n2t.net/addgene:123461 ; RRID:Addgene_123461) |
| 参考文献部分: | Improved CUT&RUN chromatin profiling tools. Meers MP, Bryson TD, Henikoff JG, Henikoff S. Elife. 2019 Jun 24;8. pii: 46314. doi: 10.7554/eLife.46314. 10.7554/eLife.46314 PubMed 31232687 |
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 pAG/MNase 3846 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses Protein A and Protein G IgG binding domains fused to
micrococcal nuclease for purposes of CUT.
ACCESSION .
VERSION .
KEYWORDS pAG/MNase
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3846)
AUTHORS Meers MP, Bryson TD, Henikoff JG, Henikoff S
TITLE Improved CUT
JOURNAL Elife. 2019 Jun 24;8. pii: 46314. doi: 10.7554/eLife.46314.
PUBMED 31232687
REFERENCE 2 (bases 1 to 3846)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..3846
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 365..1159
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(419..438)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 1029..1048
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
rep_origin 1443..2031
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1932..1951
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2185..2202
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 2319..2340
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2355..2385
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2393..2409
/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 2398..2420
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 2417..2433
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2417..2433
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS 2447..2872
/codon_start=1
/product="immunoglobulin binding domains of Staphylocuccus
aureus protein A"
/label=protein A
/translation="SLKDDPSQSANLLSEAKKLNESQAPKADNKFNKEQQNAFYEILHL
PNLNEEQRNGFIQSLKDDPSQSANLLAEAKKLNDAQAPKADNKFNKEQQNAFYEILHLP
NLTEEQRNGFIQSLKDDPSVSKEILAEAKKLNDAQAPK"
CDS 2873..3082
/codon_start=1
/product="immunoglobulin binding domain of Streptoccocus
protein G"
/label=protein G
/translation="TTYKLVINGKTLKGETTTEAVDAETAERHFKQYANDNGVDGEWTY
DDATKTFTVTEKPEVIDASELTPAV"
CDS 3101..3547
/codon_start=1
/product="micrococcal nuclease from Staphylococcus aureus"
/label=MNase
/translation="ATSTKKLHKEPATLIKAIDGDTVKLMYKGQPMTFRLLLVDTPETK
HPKKGVEKYGPEASAFTKKMVENAKKIEVEFDKGQRTDKYGRGLAYIYADGKMVNEALV
RQGLAKVAYVYKPNNTHEQHLRKSEAQAKKEKLNIWSEDNADSGQ"
CDS 3593..3610
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 3626..3652
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
primer_bind complement(3690..3707)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(3690..3706)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3699..3721)
/label=M13/pUC Forward
/note="In lacZ gene"
ORIGIN
1 cgataagcta gcttcacgct gccgcaagca ctcagggcgc aagggctgct aaaggaagcg
61 gaacacgtag aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg tcagctactg
121 ggctatctgg acaagggaaa acgcaagcgc aaagagaaag caggtagctt gcagtgggct
181 tacatggcga tagctagact gggcggtttt atggacagca agcgaaccgg aattgccagc
241 tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc
301 gccaaggatc tgatggcgca ggggatcaag atctgatcaa gagacaggat gaggatcgtt
361 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
421 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
481 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
541 actccaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
601 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
661 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
721 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
781 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
841 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcggatgcc
901 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
961 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
1021 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
1081 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
1141 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
1201 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
1261 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
1321 ttcgcccacc ccaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
1381 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
1441 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
1501 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
1561 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
1621 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
1681 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
1741 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
1801 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
1861 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
1921 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
1981 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
2041 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
2101 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
2161 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
2221 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
2281 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
2341 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
2401 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct taaaagatga
2461 cccaagccaa agtgctaacc tattgtcaga agctaaaaag ttaaatgaat ctcaagcacc
2521 gaaagcggat aacaaattca acaaagaaca acaaaatgct ttctatgaaa tcttacattt
2581 acctaactta aacgaagaac aacgcaatgg tttcatccaa agcctaaaag atgacccaag
2641 ccaaagcgct aaccttttag cagaagctaa aaagctaaat gatgctcaag caccaaaagc
2701 tgacaacaaa ttcaacaaag aacaacaaaa tgctttctat gaaattttac atttacctaa
2761 cttaactgaa gaacaacgta acggcttcat ccaaagcctt aaagacgatc cttcagtgag
2821 caaagaaatt ttagcagaag ctaaaaagct aaacgatgct caagcaccaa aaacaactta
2881 taaattagtc atcaacggga aaacgctgaa gggtgaaacc acgacagagg ccgtagatgc
2941 ggagacagcg gagcgccact ttaagcaata cgcgaatgat aacggtgtag acggcgagtg
3001 gacctacgac gacgcgacaa agacctttac cgtcacggag aaacctgagg ttatcgacgc
3061 gtctgagttg acgccagccg tagatgacga taaagaattc gcaacttcaa ctaaaaaatt
3121 acataaagaa cctgcgactt taattaaagc gattgatggt gatacggtta aattaatgta
3181 caaaggtcaa ccaatgacat tcagactatt attggtcgac acacctgaaa caaagcatcc
3241 taaaaaaggt gtagagaaat atggtcctga agcaagtgca tttaccaaaa aaatggtaga
3301 aaatgcaaag aaaattgaag tcgaattcga caaaggtcaa agaactgata aatatggacg
3361 tgggctagcg tatatttatg ctgatggaaa aatggtaaac gaagctttag ttcgtcaagg
3421 cttggctaaa gttgcttatg tttacaaacc taacaataca catgaacaac atttaagaaa
3481 aagtgaagca caagcgaaaa aagagaaatt aaatatttgg agcgaagaca acgctgattc
3541 aggtcaaggc ggtggtggct ctgggggcgg gggctcgggt ggtgggggct cacaccatca
3601 ccatcaccat ggcggtggtg gctcttaccc atacgatgtt ccagattacg cttaatgagg
3661 atccccgggt accgagctcg aattaattca ctggccgtcg ttttacaacg tcgtgactgg
3721 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
3781 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
3841 gaatgg
//
载体图谱质粒图谱pdf版和相关资料下载
载体质粒应用举例
- 上一篇：pET22b RNase A
- 下一篇：LentiCRISPRv2-mCherry
