| 质粒名称: | pMKO.1 puro |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 8452 |
| 存储实验室: | Bob Weinberg |
|
载体骨架基本信息 |
|
| 载体名称: | pMSCV |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6700 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，逆转录病毒，RNAi |
| 筛选标记: | 嘌呤霉素 |
| 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 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | EcoRI (未破坏) |
| 5' 测序引物 | pLXSN 5' |
| 3' 测序引物 | pBABE-3 |
|
文献引用方法 |
|
| 材料和方法部分: | pMKO.1 puro was a gift from Bob Weinberg (Addgene plasmid # 8452 ; http://n2t.net/addgene:8452 ; RRID:Addgene_8452) |
| 参考文献部分: | Lentivirus-delivered stable gene silencing by RNAi in primary cells. Stewart SA, Dykxhoorn DM, Palliser D, Mizuno H, Yu EY, An DS, Sabatini DM, Chen IS, Hahn WC, Sharp PA, Weinberg RA, Novina CD. RNA 2003 Apr;9(4):493-501. 10.1261/rna.2192803 PubMed 12649500 |
买家导航
pMKO.1puro载体基本信息
pMKO.1puro载体图谱质粒图谱和多克隆位点信息
pMKO.1puro载体简介
pMKO.1puro载体序列
LOCUS pMKO.1 puro 6340 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pMKO.1 puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6340)
AUTHORS Stewart SA, Dykxhoorn DM, Palliser D, Mizuno H, Yu EY, An DS,
Sabatini DM, Chen IS, Hahn WC, Sharp PA, Weinberg RA, Novina CD
TITLE Lentivirus-delivered stable gene silencing by RNAi in primary cells.
JOURNAL RNA 2003 Apr;9(4):493-501.
PUBMED 12649500
REFERENCE 2 (bases 1 to 6340)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6340
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..241
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 1..21
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 172..191
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
promoter 310..639
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 490..625
/label=SV40 ori
/note="SV40 origin of replication"
CDS 649..1248
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(649..668)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 1145..1165
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 1444..1869
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
Moloney murine leukemia virus"
primer_bind complement(1972..1994)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
promoter 2135..2464
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2315..2450
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2486..2505
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(2586..2603)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2757..3345)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2837..2856)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3516..4376)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 4139..4158
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4377..4481)
/gene="bla"
/label=AmpR promoter
primer_bind 4549..4567
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
enhancer 4667..5046
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 5047..5250
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 5201..5221
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 5251..5426
/label=5' LTR (truncated)
/note="truncated long terminal repeat from Moloney murine
sarcoma virus"
misc_feature 5489..5688
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 5889..6305
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 6235..6257
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 6273..6289
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
ORIGIN
1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
241 cgaaacaccg gtacgcgtac tagtactccc atatggaatt cgccagcaca gtggtcgacc
301 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gcttcccagc aggcagaagt
361 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
421 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
481 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
541 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
601 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttaccat gaccgagtac
661 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
721 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
781 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
841 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
901 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
961 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
1021 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
1081 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
1141 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
1201 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac
1261 gacccgcagc gcccgaccga aaggagcgca cgaccccatg catcgattag tccaatttgt
1321 taaagacagg atatcagtgg tccaggctct agttttgact caacaatatc accagctgaa
1381 gcctatagag tacgagccat agataaaata aaagatttta tttagtctcc agaaaaaggg
1441 gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa
1501 ggcatggaaa aatacataac tgagaataga gaagttcaga tcaaggtcag gaacagatgg
1561 aacagggtcg accctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg
1621 accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc
1681 tgctccccga gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat
1741 tgactgagtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt
1801 gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg
1861 tctttcattt gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac
1921 caccgggagg taagctggct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca
1981 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
2041 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg
2101 tagcgatagc ggagtgtaga tccggctgtg gaatgtgtgt cagttagggt gtggaaagtc
2161 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
2221 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
2281 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
2341 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
2401 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
2461 caaaaagctt actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
2521 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc
2581 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
2641 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
2701 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
2761 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
2821 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
2881 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
2941 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
3001 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
3061 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
3121 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
3181 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
3241 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
3301 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
3361 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
3421 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
3481 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
3541 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
3601 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
3661 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
3721 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
3781 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc
3841 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
3901 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
3961 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
4021 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
4081 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat
4141 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
4201 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
4261 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
4321 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
4381 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
4441 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
4501 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
4561 acgaggccct ttcgtcttca agaattagct tggccattgc atacgttgta tccatatcat
4621 aatatgtaca tttatattgg ctcatgtcca acattaccgc catgttgaca ttgattattg
4681 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
4741 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
4801 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
4861 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
4921 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
4981 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
5041 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
5101 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
5161 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
5221 gtacggtggg aggtctatat aagcagagct caataaaaga gcccacaacc cctcactcgg
5281 cgcgccagtc ttccgataga ctgcgtcgcc cgggtacccg tattcccaat aaagcctctt
5341 gctgtttgca tccgaatcgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg
5401 actacccacg acgggggtct ttcatttggg ggctcgtccg ggatttggag acccctgccc
5461 agggaccacc gacccaccac cgggaggtaa gctggccagc aacttatctg tgtctgtccg
5521 attgtctagt gtctatgttt gatgttatgc gcctgcgtct gtactagtta gctaactagc
5581 tctgtatctg gcggacccgt ggtggaactg acgagttctg aacacccggc cgcaaccctg
5641 ggagacgtcc cagggacttt gggggccgtt tttgtggccc gacctgagga agggagtcga
5701 tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg agacgagaac ctaaaacagt
5761 tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg aagccgcgcg tcttgtctgc
5821 tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg
5881 aaaattaggg ccagactgtt accactccct taagtttgac cttaggtcac tggaaagatg
5941 tcgagcggat cgctcacaac cagtcggtag atgtcaagaa gagacgttgg gttaccttct
6001 gctctgcaga atggccaacc tttaacgtcg gatggccgcg agacggcacc tttaaccgag
6061 acctcatcac ccaggttaag atcaaggtct tttcacctgg cccgcatgga cacccagacc
6121 aggtccccta catcgtgacc tgggaagcct tggcttttga cccccctccc tgggtcaagc
6181 cctttgtaca ccctaagcct ccgcctcctc ttcctccatc cgccccgtct ctcccccttg
6241 aacctcctcg ttcgaccccg cctcgatcct ccctttatcc agccctcact ccttctctag
6301 gcgccggaat tgaagatctg tcgacaaggt cgggcaggaa
//
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