| 出品公司: | ZYbscience |
|---|---|
| 载体名称: | pREP7 |
| 质粒类型: | 哺乳动物表达载体;附加体载体 |
| 高拷贝/低拷贝: | 高拷贝 |
| 克隆方法: | 限制性内切酶,多克隆位点 |
| 启动子: | RSV |
| 载体大小: | 9494 bp |
| 5' 测序引物及序列: | pREP Forward 5’ GCTCGATACAATAAACGCC 3’ |
| 3' 测序引物及序列: | SV40pA rev 5'GAAATTTGTGATGCTATTGC' |
| 载体标签: | -- |
| 载体抗性: | 氨苄青霉素 |
| 筛选标记: | Hygromycin |
| 克隆菌株: | DH5α 等 |
| 宿主细胞(系): | 人类细胞;灵长类细胞;犬类细胞 |
| 备注: | pREP7载体是附加体型载体,可以在基因组外自主复制,类似的载体有: |
| 产品目录号: | -- |
| 稳定性: | 稳表达 |
| 组成型/诱导型: | 组成型 |
| 病毒/非病毒: | 非病毒 |
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- 详细信息
- 文献和实验
- 技术资料
- 保存条件:
-20℃低温保存
- 保质期:
三年
- 英文名:
pREP7
- 库存:
20
- 供应商:
泽叶生物
载体基本信息
载体质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pREP7 9494 bp DNA SYN
DEFINITION pREP7
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
FEATURES Location/Qualifiers
source 1..9494
/organism="pREP7"
/mol_type="other DNA"
misc_feature 583..601
/label="pREP_fwd_primer"
misc_feature complement(717..736)
/label="EBV_rev_primer"
misc_feature complement(969..991)
/label="pGEX_3_primer"
CDS 1084..1818
/label="ORF frame 1"
/translation="MTGRPLGARNPRGGFWGSGVFGSHRRPVFVSRQYMRSMPRPSKN
HGSVCSVQSWTRPHATPKIITPTNHKPFPMGDPVPNPRGQWLWQGLPPRRWLRALGLH
PNLGGGVGKRKKRGRIGPNGVSVGYRQSASPGTEPRVYEQTTQHPCVLFCLFIAVIAR
VPSGIVSFRVSVSLPHLPYSFALGRVLGRRFPLSASTSTQPSVQTAALLRAICVRPTV
PAPDRTIASHRPCAQAASSKLPSTKL*"
terminator complement(1339..1609)
/label="TK_PA_terminator"
/translation="MTGRPLGARNPRGGFWGSGVFGSHRRPVFVSRQYMRSMPRPSKN
HGSVCSVQSWTRPHATPKIITPTNHKPFPMGDPVPNPRGQWLWQGLPPRRWLRALGLH
PNLGGGVGKRKKRGRIGPNGVSVGYRQSASPGTEPRVYEQTTQHPCVLFCLFIAVIAR
VPSGIVSFRVSVSLPHLPYSFALGRVLGRRFPLSASTSTQPSVQTAALLRAICVRPTV
PAPDRTIASHRPCAQAASSKLPSTKL*"
CDS complement(1617..2642)
/label="ORF frame 1"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQD
LPETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVY
HWQTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWS
EAMFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQ
SLVDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPS
TRPRAKE*"
gene complement(1620..2627)
/label="hygroB"
/gene="hygroB"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQD
LPETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVY
HWQTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWS
EAMFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQ
SLVDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPS
TRPRAKE*"
CDS 1884..3254
/label="ORF frame 3"
/translation="MPPLEVARLLLHTSQPRPPEEDVGDLVLGIPEHRLAPVNDRCYA
AIVRQDIVGAEIRVHEVPDFGAVLGPKHQLIESLRDGRTDGVVHHSLPVIHMGISNRA
YEITPCSVLTDSLRSEWAEPARLAKIGRSDRIHGLRDRLQNSGQFGFRQVLQRDTLCT
AGDAIGQALAEFPNVKHFRNRERGRCKVPINITIFVETIGAAIYPQDISTPSYIEAES
TRFFALRELHQVGDAVELFDQKLLDRRRGEFRLFHISLPGICGTLLTLLSGSLQGRSV
FEATRVTLICEVDLGPRRPDCICVFEFANDKTLGGVCVIIELKTCKYISSGDTASKRE
QRATGMKQGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDH
QGQLQGSLAALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAW
SRATST*"
promoter 2761..2909
/label="TATA_promoter"
/translation="MPPLEVARLLLHTSQPRPPEEDVGDLVLGIPEHRLAPVNDRCYA
AIVRQDIVGAEIRVHEVPDFGAVLGPKHQLIESLRDGRTDGVVHHSLPVIHMGISNRA
YEITPCSVLTDSLRSEWAEPARLAKIGRSDRIHGLRDRLQNSGQFGFRQVLQRDTLCT
AGDAIGQALAEFPNVKHFRNRERGRCKVPINITIFVETIGAAIYPQDISTPSYIEAES
TRFFALRELHQVGDAVELFDQKLLDRRRGEFRLFHISLPGICGTLLTLLSGSLQGRSV
FEATRVTLICEVDLGPRRPDCICVFEFANDKTLGGVCVIIELKTCKYISSGDTASKRE
QRATGMKQGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDH
QGQLQGSLAALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAW
SRATST*"
gene 2916..3254
/label="tet (852 - 1190)"
/gene="tet (852 - 1190)"
/translation="MPPLEVARLLLHTSQPRPPEEDVGDLVLGIPEHRLAPVNDRCYA
AIVRQDIVGAEIRVHEVPDFGAVLGPKHQLIESLRDGRTDGVVHHSLPVIHMGISNRA
YEITPCSVLTDSLRSEWAEPARLAKIGRSDRIHGLRDRLQNSGQFGFRQVLQRDTLCT
AGDAIGQALAEFPNVKHFRNRERGRCKVPINITIFVETIGAAIYPQDISTPSYIEAES
TRFFALRELHQVGDAVELFDQKLLDRRRGEFRLFHISLPGICGTLLTLLSGSLQGRSV
FEATRVTLICEVDLGPRRPDCICVFEFANDKTLGGVCVIIELKTCKYISSGDTASKRE
QRATGMKQGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDH
QGQLQGSLAALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAW
SRATST*"
CDS 3158..3769
/label="ORF frame 2"
/translation="MERVGMDCRRRPIPCLPPRVASRCMEPGHLDLNGSRRHLANGFT
TPRIGANQFLRRTVNAQTNPWQNISIASAISSSRTRRSKRPGTVKRPRCWRFSIGSAP
LTSITKIDAQVRGGETRQDYKDTRRFPLEAPSCALLFRPCRLPDTCPPFSLREAWRFL
IAHAVGISVRCRSFAPSWAVCTNPPFSPTAAPYPVTIVLSPTR*"
gene complement(4203..5063)
/label="Ampicillin"
/gene="Ampicillin"
/translation="MERVGMDCRRRPIPCLPPRVASRCMEPGHLDLNGSRRHLANGFT
TPRIGANQFLRRTVNAQTNPWQNISIASAISSSRTRRSKRPGTVKRPRCWRFSIGSAP
LTSITKIDAQVRGGETRQDYKDTRRFPLEAPSCALLFRPCRLPDTCPPFSLREAWRFL
IAHAVGISVRCRSFAPSWAVCTNPPFSPTAAPYPVTIVLSPTR*"
CDS complement(4203..5063)
/label="ORF frame 1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
promoter complement(5105..5133)
/label="AmpR_promoter"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
CDS 5804..6925
/label="ORF frame 2"
/translation="MDEDGEEDEDEEAEDQEPRAAQDQGQDIEMVSGDPKNVQVALAA
KGPTVEQEQEQEREGQEQEGQEGGRGRGGSGGRGRGGSGGRRGRGRERARGGSRERAR
GRGRGRGEKRPRSPSSQSSSSGSPPRRPPPGRRPFFHPVGEADYFEYHQEGGPDGEPD
VPPGAIEQGPADDPGEGPSTGPRGQGDGGRRKKGGWFGKHRGQGGSNPKFENIAEGLR
ALLARSHVERTTDEGTWVAGVFVYGGSKTSLYNLRRGTALAIPQCRLTPLSRLPFGMA
PGPGPQPGPLRESIVCYFMVFLQTHIFAEVLKDAIKDLVMTKPAPTCNIRVTVCSFDD
GVDLPPWFPPMVEGAAAEGDDGDDGDEGGDGDEGEEGQE*"
rep_origin 7229..9399
/label="EBV_origin"
/translation="MDEDGEEDEDEEAEDQEPRAAQDQGQDIEMVSGDPKNVQVALAA
KGPTVEQEQEQEREGQEQEGQEGGRGRGGSGGRGRGGSGGRRGRGRERARGGSRERAR
GRGRGRGEKRPRSPSSQSSSSGSPPRRPPPGRRPFFHPVGEADYFEYHQEGGPDGEPD
VPPGAIEQGPADDPGEGPSTGPRGQGDGGRRKKGGWFGKHRGQGGSNPKFENIAEGLR
ALLARSHVERTTDEGTWVAGVFVYGGSKTSLYNLRRGTALAIPQCRLTPLSRLPFGMA
PGPGPQPGPLRESIVCYFMVFLQTHIFAEVLKDAIKDLVMTKPAPTCNIRVTVCSFDD
GVDLPPWFPPMVEGAAAEGDDGDDGDEGGDGDEGEEGQE*"
ORIGIN
1 tctagagtcg accaattctc atgtttgaca gcttatcatc gcagatcctg agcttgtatg
61 gtgcactctc agtacaatct gctctgctgc cgcatagtta agccagtatc tgctccctgc
121 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
181 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
241 tgtacgggcc agatatacgc gtatctgagg ggactagggt gtgtttaggc gcccagcggg
301 gcttcggttg tacgcggtta ggagtcccct caggatatag tagtttcgct tttgcatagg
361 gagggggaaa tgtagtctta tgcaatacac ttgtagtctt gcaacatggt aacgatgagt
421 tagcaacatg ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg
481 tggtacgatc gtgccttatt aggaaggcaa cagacaggtc tgacatggat tggacgaacc
541 actgaattcc gcattgcaga gataattgta tttaagtgcc tagctcgata caataaacgc
601 catttgacca ttcaccacat tggtgtgcac ctccaagctg ggtaccagct gctagcaagc
661 ttgctagcgg ccgctcgagg ccggcaaggc cggatccaga catgataaga tacattgatg
721 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
781 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
841 gcattcattt tatgtttcag gttcaggggg aggtggggag gttttttaaa gcaagtaaaa
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文献和实验Yale RNA prep (Rebecca D. Burdine, Michael J. Stern [modified by S.M. Hettenbach]) Add 8 ml of TRIZOL to 2 ml packed worms in 15 ml centrifuge tube. Vortex and invert tube to solubilize and lyse worms for at least 10 min.
A quick RNA mini-prep for Neurospora mycelial cultures
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1.常用载体分类:2,常用载体元件介绍:(1) 启动子:启动子是位于结构基因5'端上游的DNA序列,能活化RNA聚合酶,使之与模板DNA准确的结合并具有转录起始的特异性。启动子主要分为广谱启动子和特异性启动子。广谱启动子为在各种组织、细胞上广泛表达的启动子;特异性启动子为在特异性组织或者细胞类型上表达的启动子。常用广谱启动子有CAG,CMV,EF1a,PGK等,还包含启动RNA的III型启动子U6和H1启动子;常用特异性启动子如成熟神经元特异性启动子hSyn,投射神经元特异性启动
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