碧云天生物技术-pCMV-N-Flag(pCMV-N-Flag质粒)(D2722-100μg)

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pCMV-N-Flag

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产品编号: D2722-100μg

产品包装:100μg

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产品编号	产品名称	产品包装	产品价格
D2722-1μg	pCMV-N-Flag	1μg	888.00元
D2722-100μg	pCMV-N-Flag	100μg	1169.00元

pCMV-N-Flag是碧云天自行研发的用于在哺乳动物细胞中表达N端和Flag tag(Flag标签)融合的目的蛋白的表达质粒。含有CMV启动子可以高效启动目的蛋白在细胞中的表达；在多克隆位点的5'端含有一个可以编码Flag标签的序列，因此可以表达出含有Flag标签的融合蛋白，可以方便地使用抗Flag的抗体来识别目的蛋白，有利于目的蛋白检测和分离纯化。质粒为卡那霉素抗性。转染细胞后，可使用G418筛选稳定表达目的蛋白的细胞株。
pCMV-N-Flag质粒的主要信息如下：

Feature Nucleotide	Position
CMV promoter	1-602
T3 promoter and T3 primer binding site	620-639
Flag tag	679-702
multiple cloning site	705-778
T7 promoter and T7 primer binding site	821-842
SV40 polyA signal	854-1237
f1 origin of ss-DNA replication	1375-1681
bla promoter	1706-1830
SV40 promoter	1850-2188
neomycin/kanamycin resistance ORF	2223-3014
HSV-thymidine kinase(TK)polyA signal	3015-3473
pUC origin	3602-4269

pCMV-N-Flag质粒的图谱如下：

pCMV-N-Flag质粒的详细图谱见说明书：https://www.beyotime.com/Manual/D2722 pCMV-N-Flag.pdf 。

pCMV-N-Flag中没有的酶切位点(Restriction enzymes that do not cut pCMV-N-Flag)包括：

Afl II	Age I	Ahd I	Asc I	Bbs I	Bbv II	Blp I
Bsg I	BsiW I	BsmB I	BspM II	BsrG I	BssH II	Bst1107 I
BstE II	Ear I	Eco47 III	Eco72 I	EcoN I	Esp I	Fse I
Nru I	PflM I	Pme I	Pml I	PpuM I	Psp1406 I	Sap I
Sca I	Spl I

pCMV-N-Flag中的单酶切位点(Restriction enzymes that cut pCMV-N-Flag once)包括：

Nde I	CA`TA,TG	241	Pvu I	CG,AT`CG	855
SnaB I	TAC\|GTA	347	Bcl I	T`GATC,A	1009
Nhe I	G`CTAG,C	598	Mun I	C`AATT,G	1102
Sac I	G,AGCT`C	656	Hpa I	GTT\|AAC	1115
Sac II	CC,GC`GG	663	Mlu I	A`CGCG,T	1238
BstX I	CCAN,NNNN`NTGG	664	Dra III	CAC,NNN`GTG	1468
Not I	GC`GGCC,GC	669	Sfi I	GGCCN,NNN`NGGCC	2127
PspA I	C`CCGG,G	706	BseR I	GAGGAG 16/14	2170
Xma I	C`CCGG,G	706	Stu I	AGG\|CCT	2173
Srf I	GCCC\|GGGC	708	Cla I	AT`CG,AT	2192
Sma I	CCC\|GGG	708	Kas I	G`GCGC,C	2351
BamH I	G`GATC,C	713	Nar I	GG`CG,CC	2352
Hind III	A`AGCT,T	719	Ehe I	GGC\|GCC	2353
Pst I	C,TGCA`G	729	Bbe I	G,GCGC`C	2355
EcoR I	G`AATT,C	731	Msc I	TGG\|CCA	2434
EcoR V	GAT\|ATC	739	Tth111 I	GACN`N,NGTC	2470
Sal I	G`TCGA,C	743	BsrD I	GCAATG, 8	2585
Acc I	GT`MK,AC	744	Bsp1286 I	G,DGCH`C	2655
Bgl II	A`GATC,T	749	Rsr II	CG`GWC,CG	2868
PaeR7 I	C`TCGA,G	755	BsiC I	TT`CG,AA	3034
Xho I	C`TCGA,G	755	BstB I	TT`CG,AA	3034
Xba I	T`CTAG,A	761	Bsa I	GGTCTC 7/11	3341
Spe I	A`CTAG,T	767	HgiE II	ACCNNNNNNGGT-1/13	3681
Bsp120 I	G`GGCC,C	773	ApaL I	G`TGCA,C	3956
Apa I	G,GGCC`C	777

pCMV-N-Flag质粒中对于插入片段进行测序时，推荐使用的正向测序引物T3和反向测序引物T7的序列如下：
T3 primer(620-639): 5′AATTAACCCTCACTAAAGGG 3′
T7 primer(821-842): 5′GTAATACGACTCACTATAGGGC 3′
pCMV-N-Flag的全序列信息：D2722 pCMV-N-Flag-seq.txt
包装清单：

产品编号	产品名称	包装
D2722-1μg	pCMV-N-Flag	1μg
D2722-001μg	pCMV-N-Flag	100μg
—	说明书	1份

保存条件：
-20℃保存。
注意事项：
本质粒未经碧云天书面许可不得用于任何商业用途，也不得移交给订货人所在实验室外的任何个人或单位。
本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。
为了您的安全和健康，请穿实验服并戴一次性手套操作。

使用说明：
1. 首次使用时请先取少量本质粒转化大肠杆菌，进行质粒小量、中量或大量抽提后再用于后续用途。抽提获得的质粒可以通过酶切电泳进行鉴定，或通过测序进行鉴定。
2. 100μg包装的本产品质粒浓度为0.1μg/μl，共1ml。可以直接用于酶切或者转染细胞。
3. pCMV-N-Flag质粒在其多克隆位点适当酶切后可以插入待表达的目的基因，需注意插入基因片段和tag之间的读码框要一致，即需要避免发生移码突变。构建的质粒可以用常规方法转染细胞。

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