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- 2025年07月13日
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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
-20℃
- 保质期：
6个月
- 英文名：
pLenti4/V5-DEST
- 库存：
大量
- 供应商：
钰博生物
- 规格：
1ug/5ug
pLenti4/V5-DEST
载体基本信息
| 出品公司: | Ybscience |
|---|---|
| 载体名称: | pLenti4/V5-Dest |
| 质粒类型: | Gateway载体；慢病毒载体 |
| 高拷贝/低拷贝: | -- |
| 启动子: | CMV |
| 克隆方法: | Gateway |
| 载体大小: | 8634bp |
| 5' 测序引物及序列: | CMVPro Fwd: 5'd[CGCAAATGGGCGGTAGGCGTG]3' |
| 3' 测序引物及序列: | -- |
| 载体标签: | V5 |
| 载体抗性: | 氨苄青霉素，氯霉素（仅空载体） |
| 筛选标记: | 博来霉素Zeocin |
| 备注: | 克隆菌株 E.coli Stbl3，DB3.1 |
| 稳定性: | 稳表达 |
| 组成型: | 组成型 |
| 病毒/非病毒: | 慢病毒 |
载体质粒图谱和多克隆位点信息
载体简介
The pLenti4/V5-DEST Gateway Vector is a Gateway-adapted ViraPower lentiviral expression vector for lentiviral-based expression of a target gene in dividing and non-dividing mammalian cells. The vector has the CMV promoter for driving constitutive expression of the target gene and the Zeocin selection marker for stable selection in mammalian cells. Advantages Lentivirus based expression of a target gene in dividing and non-dividing mammalian cells Key Features Flexible and versatile Gateway recombination cloning technology Constitutive high expression with CMV promoter Zeocin selection marker for stable selection C terminal V5 tag for quick detection
载体序列
LOCUS pLenti4/V5-DEST Gateway (ViraPower) 8634 bp DNA SYN
DEFINITION pLenti4/V5-DEST Gateway (ViraPower)
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
FEATURES Location/Qualifiers
source 1..8634
/organism="pLenti4/V5-DEST Gateway (ViraPower)"
/mol_type="other DNA"
misc_feature 230..410
/label="HIV-1_5_LTR"
misc_feature 230..410
/label="truncHIV-1_3_LTR"
misc_feature 521..565
/label="HIV-1_psi_pack"
CDS 953..1801
/label="ORF frame 2"
misc_feature 1075..1308
/label="RRE"
promoter 1813..2317
/label="CMV_immearly_promoter"
misc_feature 1820..2107
/label="CAG_enhancer"
misc_feature 2274..2294
/label="CMV_fwd_primer"
promoter 2275..2344
/label="CMV_promoter"
misc_feature 2318..2337
/label="pCEP_fwd_primer"
misc_feature 2320..2344
/label="LNCX_primer"
misc_feature 2440..2564
/label="attR1"
misc_feature complement(2464..2564)
/label="attR2"
promoter 2589..2612
/label="lac_promoter"
gene 2673..3332
/label="CAT/CamR"
/gene="CAT/CamR"
CDS 2673..3332
/label="ORF frame 3"
misc_feature 3674..3979
/label="ccdB"
misc_feature complement(4020..4127)
/label="attR1"
misc_feature 4020..4120
/label="attR2"
misc_feature complement(4275..4295)
/label="pBABE_3_primer"
misc_feature complement(4281..4497)
/label="SV40_enhancer"
promoter 4293..4562
/label="SV40_promoter"
rep_origin 4461..4538
/label="SV40_origin"
misc_feature 4523..4542
/label="SV40pro_F_primer"
promoter 4632..4687
/label="EM7_promoter"
gene 4688..5059
/label="bleo"
/gene="bleo"
misc_feature 4688..5062
/label="sh_ble"
misc_feature 5138..5153
/label="cPPT"
misc_feature 5138..5159
/label="U3PPT"
misc_feature 5155..5207
/label="delta_U3"
misc_feature 5208..5388
/label="HIV-1_5_LTR"
misc_feature 5208..5388
/label="truncHIV-1_3_LTR"
terminator 5463..5582
/label="SV40_PA_terminator"
misc_feature 5551..5570
/label="EBV_rev_primer"
rep_origin 5600..5698
/label="SV40_origin"
misc_feature 5683..5702
/label="SV40pro_F_primer"
promoter complement(5777..5795)
/label="T7_promoter"
promoter complement(5805..5821)
/label="M13_forward20_primer"
misc_feature complement(5814..5836)
/label="M13_pUC_fwd_primer"
misc_feature 5814..5962
/label="lacZ_a"
rep_origin 5980..6286
/label="f1_origin"
promoter 6479..6507
/label="AmpR_promoter"
gene 6549..7409
/label="Ampicillin"
/gene="Ampicillin"
CDS 6549..7409
/label="ORF frame 3"
rep_origin 7564..8183
/label="pBR322_origin"
promoter 8492..8521
/label="lac_promoter"
misc_feature 8535..8557
/label="M13_pUC_rev_primer"
promoter 8556..8574
/label="M13_reverse_primer"
promoter 8591..8610
/label="T3_promoter"
ORIGIN
1 AATGTAGTCT TATGCAATAC TCTTGTAGTC TTGCAACATG GTAACGATGA GTTAGCAACA
61 TGCCTTACAA GGAGAGAAAA AGCACCGTGC ATGCCGATTG GTGGAAGTAA GGTGGTACGA
121 TCGTGCCTTA TTAGGAAGGC AACAGACGGG TCTGACATGG ATTGGACGAA CCACTGAATT
181 GCCGCATTGC AGAGATATTG TATTTAAGTG CCTAGCTCGA TACATAAACG GGTCTCTCTG
241 GTTAGACCAG ATCTGAGCCT GGGAGCTCTC TGGCTAACTA GGGAACCCAC TGCTTAAGCC
301 TCAATAAAGC TTGCCTTGAG TGCTTCAAGT AGTGTGTGCC CGTCTGTTGT GTGACTCTGG
361 TAACTAGAGA TCCCTCAGAC CCTTTTAGTC AGTGTGGAAA ATCTCTAGCA GTGGCGCCCG
421 AACAGGGACT TGAAAGCGAA AGGGAAACCA GAGGAGCTCT CTCGACGCAG GACTCGGCTT
481 GCTGAAGCGC GCACGGCAAG AGGCGAGGGG CGGCGACTGG TGAGTACGCC AAAAATTTTG
541 ACTAGCGGAG GCTAGAAGGA GAGAGATGGG TGCGAGAGCG TCAGTATTAA GCGGGGGAGA
601 ATTAGATCGC GATGGGAAAA AATTCGGTTA AGGCCAGGGG GAAAGAAAAA ATATAAATTA
661 AAACATATAG TATGGGCAAG CAGGGAGCTA GAACGATTCG CAGTTAATCC TGGCCTGTTA
721 GAAACATCAG AAGGCTGTAG ACAAATACTG GGACAGCTAC AACCATCCCT TCAGACAGGA
781 TCAGAAGAAC TTAGATCATT ATATAATACA GTAGCAACCC TCTATTGTGT GCATCAAAGG
841 ATAGAGATAA AAGACACCAA GGAAGCTTTA GACAAGATAG AGGAAGAGCA AAACAAAAGT
901 AAGACCACCG CACAGCAAGC GGCCGCTGAT CTTCAGACCT GGAGGAGGAG ATATGAGGGA
961 CAATTGGAGA AGTGAATTAT ATAAATATAA AGTAGTAAAA ATTGAACCAT TAGGAGTAGC
1021 ACCCACCAAG GCAAAGAGAA GAGTGGTGCA GAGAGAAAAA AGAGCAGTGG GAATAGGAGC
1081 TTTGTTCCTT GGGTTCTTGG GAGCAGCAGG AAGCACTATG GGCGCAGCGT CAATGACGCT
1141 GACGGTACAG GCCAGACAAT TATTGTCTGG TATAGTGCAG CAGCAGAACA ATTTGCTGAG
1201 GGCTATTGAG GCGCAACAGC ATCTGTTGCA ACTCACAGTC TGGGGCATCA AGCAGCTCCA
1261 GGCAAGAATC CTGGCTGTGG AAAGATACCT AAAGGATCAA CAGCTCCTGG GGATTTGGGG
1321 TTGCTCTGGA AAACTCATTT GCACCACTGC TGTGCCTTGG AATGCTAGTT GGAGTAATAA
1381 ATCTCTGGAA CAGATTTGGA ATCACACGAC CTGGATGGAG TGGGACAGAG AAATTAACAA
1441 TTACACAAGC TTAATACACT CCTTAATTGA AGAATCGCAA AACCAGCAAG AAAAGAATGA
1501 ACAAGAATTA TTGGAATTAG ATAAATGGGC AAGTTTGTGG AATTGGTTTA ACATAACAAA
1561 TTGGCTGTGG TATATAAAAT TATTCATAAT GATAGTAGGA GGCTTGGTAG GTTTAAGAAT
1621 AGTTTTTGCT GTACTTTCTA TAGTGAATAG AGTTAGGCAG GGATATTCAC CATTATCGTT
1681 TCAGACCCAC CTCCCAACCC CGAGGGGACC CGACAGGCCC GAAGGAATAG AAGAAGAAGG
1741 TGGAGAGAGA GACAGAGACA GATCCATTCG ATTAGTGAAC GGATCTCGAC GGTATCGATA
1801 AGCTTGGGAG TTCCGCGTTA CATAACTTAC GGTAAATGGC CCGCCTGGCT GACCGCCCAA
1861 CGACCCCCGC CCATTGACGT CAATAATGAC GTATGTTCCC ATAGTAACGC CAATAGGGAC
1921 TTTCCATTGA CGTCAATGGG TGGAGTATTT ACGGTAAACT GCCCACTTGG CAGTACATCA
1981 AGTGTATCAT ATGCCAAGTA CGCCCCCTAT TGACGTCAAT GACGGTAAAT GGCCCGCCTG
2041 GCATTATGCC CAGTACATGA CCTTATGGGA CTTTCCTACT TGGCAGTACA TCTACGTATT
2101 AGTCATCGCT ATTACCATGG TGATGCGGTT TTGGCAGTAC ATCAATGGGC GTGGATAGCG
2161 GTTTGACTCA CGGGGATTTC CAAGTCTCCA CCCCATTGAC GTCAATGGGA GTTTGTTTTG
2221 GCACCAAAAT CAACGGGACT TTCCAAAATG TCGTAACAAC TCCGCCCCAT TGACGCAAAT
2281 GGGCGGTAGG CGTGTACGGT GGGAGGTCTA TATAAGCAGA GCTCGTTTAG TGAACCGTCA
2341 GATCGCCTGG AGACGCCATC CACGCTGTTT TGACCTCCAT AGAAGACACC GACTCTAGAG
2401 GATCCACTAG TCCAGTGTGG TGGAATTCTG CAGATATCAA CAAGTTTGTA CAAAAAAGCT
2461 GAACGAGAAA CGTAAAATGA TATAAATATC AATATATTAA ATTAGATTTT GCATAAAAAA
2521 CAGACTACAT AATACTGTAA AACACAACAT ATCCAGTCAC TATGGCGGCC GCATTAGGCA
2581 CCCCAGGCTT TACACTTTAT GCTTCCGGCT CGTATAATGT GTGGATTTTG AGTTAGGATC
2641 CGGCGAGATT TTCAGGAGCT AAGGAAGCTA AAATGGAGAA AAAAATCACT GGATATACCA
2701 CCGTTGATAT ATCCCAATGG CATCGTAAAG AACATTTTGA GGCATTTCAG TCAGTTGCTC
2761 AATGTACCTA TAACCAGACC GTTCAGCTGG ATATTACGGC CTTTTTAAAG ACCGTAAAGA
2821 AAAATAAGCA CAAGTTTTAT CCGGCCTTTA TTCACATTCT TGCCCGCCTG ATGAATGCTC
2881 ATCCGGAATT CCGTATGGCA ATGAAAGACG GTGAGCTGGT GATATGGGAT AGTGTTCACC
2941 CTTGTTACAC CGTTTTCCAT GAGCAAACTG AAACGTTTTC ATCGCTCTGG AGTGAATACC
3001 ACGACGATTT CCGGCAGTTT CTACACATAT ATTCGCAAGA TGTGGCGTGT TACGGTGAAA
3061 ACCTGGCCTA TTTCCCTAAA GGGTTTATTG AGAATATGTT TTTCGTCTCA GCCAATCCCT
3121 GGGTGAGTTT CACCAGTTTT GATTTAAACG TGGCCAATAT GGACAACTTC TTCGCCCCCG
3181 TTTTCACCAT GGGCAAATAT TATACGCAAG GCGACAAGGT GCTGATGCCG CTGGCGATTC
3241 AGGTTCATCA TGCCGTCTGT GATGGCTTCC ATGTCGGCAG AATGCTTAAT GAATTACAAC
3301 AGTACTGCGA TGAGTGGCAG GGCGGGGCGT AAAGATCTGG ATCCGGCTTA CTAAAAGCCA
3361 GATAACAGTA TGCGTATTTG CGCGCTGATT TTTGCGGTAT AAGAATATAT ACTGATATGT
3421 ATACCCGAAG TATGTCAAAA AGAGGTGTGC TATGAAGCAG CGTATTACAG TGACAGTTGA
3481 CAGCGACAGC TATCAGTTGC TCAAGGCATA TATGATGTCA ATATCTCCGG TCTGGTAAGC
3541 ACAACCATGC AGAATGAAGC CCGTCGTCTG CGTGCCGAAC GCTGGAAAGC GGAAAATCAG
3601 GAAGGGATGG CTGAGGTCGC CCGGTTTATT GAAATGAACG GCTCTTTTGC TGACGAGAAC
3661 AGGGACTGGT GAAATGCAGT TTAAGGTTTA CACCTATAAA AGAGAGAGCC GTTATCGTCT
3721 GTTTGTGGAT GTACAGAGTG ATATTATTGA CACGCCCGGG CGACGGATGG TGATCCCCCT
3781 GGCCAGTGCA CGTCTGCTGT CAGATAAAGT CTCCCGTGAA CTTTACCCGG TGGTGCATAT
3841 CGGGGATGAA AGCTGGCGCA TGATGACCAC CGATATGGCC AGTGTGCCGG TCTCCGTTAT
3901 CGGGGAAGAA GTGGCTGATC TCAGCCACCG CGAAAATGAC ATCAAAAACG CCATTAACCT
3961 GATGTTCTGG GGAATATAAA TGTCAGGCTC CGTTATACAC AGCCAGTCTG CAGGTCGACC
4021 ATAGTGACTG GATATGTTGT GTTTTACAGT ATTATGTAGT CTGTTTTTTA TGCAAAATCT
4081 AATTTAATAT ATTGATATTT ATATCATTTT ACGTTTCTCG TTCAGCTTTC TTGTACAAAG
4141 TGGTTGATAT CCAGCACAGT GGCGGCCGCT CGAGTCTAGA GGGCCCGCGG TTCGAAGGTA
4201 AGCCTATCCC TAACCCTCTC CTCGGTCTCG ATTCTACGCG TACCGGTTAG TAATGAGTTT
4261 GGAATTAATT CTGTGGAATG TGTGTCAGTT AGGGTGTGGA AAGTCCCCAG GCTCCCCAGG
4321 CAGGCAGAAG TATGCAAAGC ATGCATCTCA ATTAGTCAGC AACCAGGTGT GGAAAGTCCC
4381 CAGGCTCCCC AGCAGGCAGA AGTATGCAAA GCATGCATCT CAATTAGTCA GCAACCATAG
4441 TCCCGCCCCT AACTCCGCCC ATCCCGCCCC TAACTCCGCC CAGTTCCGCC CATTCTCCGC
4501 CCCATGGCTG ACTAATTTTT TTTATTTATG CAGAGGCCGA GGCCGCCTCT GCCTCTGAGC
4561 TATTCCAGAA GTAGTGAGGA GGCTTTTTTG GAGGCCTAGG CTTTTGCAAA AAGCTCCCCC
4621 TGTTGACAAT TAATCATCGG CATAGTATAT CGGCATAGTA TAATACGACA AGGTGAGGAA
4681 CTAAACCATG GCCAAGTTGA CCAGTGCCGT TCCGGTGCTC ACCGCGCGCG ACGTCGCCGG
4741 AGCGGTCGAG TTCTGGACCG ACCGGCTCGG GTTCTCCCGG GACTTCGTGG AGGACGACTT
4801 CGCCGGTGTG GTCCGGGACG ACGTGACCCT GTTCATCAGC GCGGTCCAGG ACCAGGTGGT
4861 GCCGGACAAC ACCCTGGCCT GGGTGTGGGT GCGCGGCCTG GACGAGCTGT ACGCCGAGTG
4921 GTCGGAGGTC GTGTCCACGA ACTTCCGGGA CGCCTCCGGG CCGGCCATGA CCGAGATCGG
4981 CGAGCAGCCG TGGGGGCGGG AGTTCGCCCT GCGCGACCCG GCCGGCAACT GCGTGCACTT
5041 CGTGGCCGAG GAGCAGGACT GACACGTGCT ACGAGATTTA AATGGTACCT TTAAGACCAA
5101 TGACTTACAA GGCAGCTGTA GATCTTAGCC ACTTTTTAAA AGAAAAGGGG GGACTGGAAG
5161 GGCTAATTCA CTCCCAACGA AGACAAGATC TGCTTTTTGC TTGTACTGGG TCTCTCTGGT
5221 TAGACCAGAT CTGAGCCTGG GAGCTCTCTG GCTAACTAGG GAACCCACTG CTTAAGCCTC
5281 AATAAAGCTT GCCTTGAGTG CTTCAAGTAG TGTGTGCCCG TCTGTTGTGT GACTCTGGTA
5341 ACTAGAGATC CCTCAGACCC TTTTAGTCAG TGTGGAAAAT CTCTAGCAGT AGTAGTTCAT
5401 GTCATCTTAT TATTCAGTAT TTATAACTTG CAAAGAAATG AATATCAGAG AGTGAGAGGA
5461 ACTTGTTTAT TGCAGCTTAT AATGGTTACA AATAAAGCAA TAGCATCACA AATTTCACAA
5521 ATAAAGCATT TTTTTCACTG CATTCTAGTT GTGGTTTGTC CAAACTCATC AATGTATCTT
5581 ATCATGTCTG GCTCTAGCTA TCCCGCCCCT AACTCCGCCC ATCCCGCCCC TAACTCCGCC
5641 CAGTTCCGCC CATTCTCCGC CCCATGGCTG ACTAATTTTT TTTATTTATG CAGAGGCCGA
5701 GGCCGCCTCG GCCTCTGAGC TATTCCAGAA GTAGTGAGGA GGCTTTTTTG GAGGCCTAGG
5761 GACGTACCCA ATTCGCCCTA TAGTGAGTCG TATTACGCGC GCTCACTGGC CGTCGTTTTA
5821 CAACGTCGTG ACTGGGAAAA CCCTGGCGTT ACCCAACTTA ATCGCCTTGC AGCACATCCC
5881 CCTTTCGCCA GCTGGCGTAA TAGCGAAGAG GCCCGCACCG ATCGCCCTTC CCAACAGTTG
5941 CGCAGCCTGA ATGGCGAATG GGACGCGCCC TGTAGCGGCG CATTAAGCGC GGCGGGTGTG
6001 GTGGTTACGC GCAGCGTGAC CGCTACACTT GCCAGCGCCC TAGCGCCCGC TCCTTTCGCT
6061 TTCTTCCCTT CCTTTCTCGC CACGTTCGCC GGCTTTCCCC GTCAAGCTCT AAATCGGGGG
6121 CTCCCTTTAG GGTTCCGATT TAGTGCTTTA CGGCACCTCG ACCCCAAAAA ACTTGATTAG
6181 GGTGATGGTT CACGTAGTGG GCCATCGCCC TGATAGACGG TTTTTCGCCC TTTGACGTTG
6241 GAGTCCACGT TCTTTAATAG TGGACTCTTG TTCCAAACTG GAACAACACT CAACCCTATC
6301 TCGGTCTATT CTTTTGATTT ATAAGGGATT TTGCCGATTT CGGCCTATTG GTTAAAAAAT
6361 GAGCTGATTT AACAAAAATT TAACGCGAAT TTTAACAAAA TATTAACGCT TACAATTTAG
6421 GTGGCACTTT TCGGGGAAAT GTGCGCGGAA CCCCTATTTG TTTATTTTTC TAAATACATT
6481 CAAATATGTA TCCGCTCATG AGACAATAAC CCTGATAAAT GCTTCAATAA TATTGAAAAA
6541 GGAAGAGTAT GAGTATTCAA CATTTCCGTG TCGCCCTTAT TCCCTTTTTT GCGGCATTTT
6601 GCCTTCCTGT TTTTGCTCAC CCAGAAACGC TGGTGAAAGT AAAAGATGCT GAAGATCAGT
6661 TGGGTGCACG AGTGGGTTAC ATCGAACTGG ATCTCAACAG CGGTAAGATC CTTGAGAGTT
6721 TTCGCCCCGA AGAACGTTTT CCAATGATGA GCACTTTTAA AGTTCTGCTA TGTGGCGCGG
6781 TATTATCCCG TATTGACGCC GGGCAAGAGC AACTCGGTCG CCGCATACAC TATTCTCAGA
6841 ATGACTTGGT TGAGTACTCA CCAGTCACAG AAAAGCATCT TACGGATGGC ATGACAGTAA
6901 GAGAATTATG CAGTGCTGCC ATAACCATGA GTGATAACAC TGCGGCCAAC TTACTTCTGA
6961 CAACGATCGG AGGACCGAAG GAGCTAACCG CTTTTTTGCA CAACATGGGG GATCATGTAA
7021 CTCGCCTTGA TCGTTGGGAA CCGGAGCTGA ATGAAGCCAT ACCAAACGAC GAGCGTGACA
7081 CCACGATGCC TGTAGCAATG GCAACAACGT TGCGCAAACT ATTAACTGGC GAACTACTTA
7141 CTCTAGCTTC CCGGCAACAA TTAATAGACT GGATGGAGGC GGATAAAGTT GCAGGACCAC
7201 TTCTGCGCTC GGCCCTTCCG GCTGGCTGGT TTATTGCTGA TAAATCTGGA GCCGGTGAGC
7261 GTGGGTCTCG CGGTATCATT GCAGCACTGG GGCCAGATGG TAAGCCCTCC CGTATCGTAG
7321 TTATCTACAC GACGGGGAGT CAGGCAACTA TGGATGAACG AAATAGACAG ATCGCTGAGA
7381 TAGGTGCCTC ACTGATTAAG CATTGGTAAC TGTCAGACCA AGTTTACTCA TATATACTTT
7441 AGATTGATTT AAAACTTCAT TTTTAATTTA AAAGGATCTA GGTGAAGATC CTTTTTGATA
7501 ATCTCATGAC CAAAATCCCT TAACGTGAGT TTTCGTTCCA CTGAGCGTCA GACCCCGTAG
7561 AAAAGATCAA AGGATCTTCT TGAGATCCTT TTTTTCTGCG CGTAATCTGC TGCTTGCAAA
7621 CAAAAAAACC ACCGCTACCA GCGGTGGTTT GTTTGCCGGA TCAAGAGCTA CCAACTCTTT
7681 TTCCGAAGGT AACTGGCTTC AGCAGAGCGC AGATACCAAA TACTGTTCTT CTAGTGTAGC
7741 CGTAGTTAGG CCACCACTTC AAGAACTCTG TAGCACCGCC TACATACCTC GCTCTGCTAA
7801 TCCTGTTACC AGTGGCTGCT GCCAGTGGCG ATAAGTCGTG TCTTACCGGG TTGGACTCAA
7861 GACGATAGTT ACCGGATAAG GCGCAGCGGT CGGGCTGAAC GGGGGGTTCG TGCACACAGC
7921 CCAGCTTGGA GCGAACGACC TACACCGAAC TGAGATACCT ACAGCGTGAG CTATGAGAAA
7981 GCGCCACGCT TCCCGAAGGG AGAAAGGCGG ACAGGTATCC GGTAAGCGGC AGGGTCGGAA
8041 CAGGAGAGCG CACGAGGGAG CTTCCAGGGG GAAACGCCTG GTATCTTTAT AGTCCTGTCG
8101 GGTTTCGCCA CCTCTGACTT GAGCGTCGAT TTTTGTGATG CTCGTCAGGG GGGCGGAGCC
8161 TATGGAAAAA CGCCAGCAAC GCGGCCTTTT TACGGTTCCT GGCCTTTTGC TGGCCTTTTG
8221 CTCACATGTT CTTTCCTGCG TTATCCCCTG ATTCTGTGGA TAACCGTATT ACCGCCTTTG
8281 AGTGAGCTGA TACCGCTCGC CGCAGCCGAA CGACCGAGCG CAGCGAGTCA GTGAGCGAGG
8341 AAGCGGAAGA GCGCCCAATA CGCAAACCGC CTCTCCCCGC GCGTTGGCCG ATTCATTAAT
8401 GCAGCTGGCA CGACAGGTTT CCCGACTGGA AAGCGGGCAG TGAGCGCAAC GCAATTAATG
8461 TGAGTTAGCT CACTCATTAG GCACCCCAGG CTTTACACTT TATGCTTCCG GCTCGTATGT
8521 TGTGTGGAAT TGTGAGCGGA TAACAATTTC ACACAGGAAA CAGCTATGAC CATGATTACG
8581 CCAAGCGCGC AATTAACCCT CACTAAAGGG AACAAAAGCT GGAGCTGCAA GCTT
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文献和实验相关实验
用293FT细胞系和相关的ViraPower™ 试剂制备的高滴度的慢病毒储备液，进行有效的和可控制的导入。一旦制备好了慢病毒储备液，就可以转染细胞以观测RNAi反应（图9）。pLenti6/BLOCK-iT™-DEST 慢病毒 RNAi 载体（ pLenti6/BLOCK-iT™-DEST Lentiviral RNAi vector）提供能够长期观察稳定RNAi效应的能力。这个载体包含包装U6RNAi框到病毒所需的全部的元件，因而你能够转染shRNA到正在分裂、难于转染和不分裂的细胞中（图10
Inducible shRNA Lentiviral Vectors
-- pENTR™ -gus (50 ng/µl) -- 2 µl pLenti4/BLOCK-iT™ -DEST vector (150 ng/µl) 1 µl 1 µl
: 2 � 3 µg/µl Ø equimolar amount of sense and antisense oligo : 400 pmol each (approx. 5 µg): 4 µl Ø 20 µl 10x Buffer B (Roche) Ø A.dest . ad 200 µl (172 µl) Ø
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pLenti4/V5-DEST
¥800 - 1200
