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pEGFP-N1哺乳细胞绿色荧光载体质粒

货号：kl-zl-1068-01

货号	产品名称	规格	库存	价格	数量	购买
kl-zl-1068-01	pEGFP-N1哺乳细胞绿色荧光载体质粒	NA		立即咨询	+ -

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pEGFP-N1质粒货号：kl-zl-1068 规格：20ul

启动子：	CMV
复制子：	pUC ori，f1 ori
终止子：	SV40 poly(A) signal
质粒分类：	哺乳细胞，荧光蛋白报告载体
质粒大小：	4733bp
原核抗性：	Kan
筛选标记：	Neo
克隆菌株：	DH5α
培养条件：	37℃，有氧 LB
表达宿主：	哺乳细胞
诱导方式：	无须诱导，瞬时表达
5'测序引物：	CMV-F：CGCAAATGGGCGGTAGGCGTG
3'测序引物：	pEGFP-N： CGTCGCCGTCCAGCTCGACCAG

质粒简介：
pEGFP-N1 encodes a red-shifted variant of wild-type GFP which has been optimized for brighter fluorescence and higher expression in mammalian cells. (Excitation maximum = 488 nm; emission maximum = 507 nm.) pEGFP-N1 encodes the GFPmut1 variant which contains the double-amino-acid substitution of Phe-64 to Leu and Ser-65 to Thr. The coding sequence of the EGFP gene contains more than 190 silent base changes which correspond to human codon-usage preferences. Sequences flanking EGFP have been converted to a Kozak consensus translation initiation site to further increase the translation efficiency in eukaryotic cells. The MCS in pEGFP-N1 is between the immediate early promoter of CMV and the EGFP coding sequences. Genes cloned into the MCS will be expressed as fusions to the N-terminus of EGFP if they are in the same reading frame as EGFP and there are no intervening stop codons. SV40 polyadenylation signals downstream of the EGFP gene direct proper processing of the 3' end of the EGFP mRNA. The vector backbone also contains an SV40 origin for replication in mammalian cells expressing the SV40 T antigen. A neomycin-resistance cassette, consisting of the SV40 early promoter, the neomycin/kanamycin resistance gene of Tn5, and polyadenylation signals from the Herpes simplex virus thymidine kinase (HSV TK) gene, allows stably transfected eukaryotic cells to be selected using G418. A bacterial promoter upstream of this cassette expresses kanamycin resistance in E. coli. The pEGFP-N1 backbone also provides a pUC origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.
Fusions to the N terminus of EGFP retain the fluorescent properties of the native protein allowing the localization of the fusion protein in vivo . The target gene should be cloned into pEGFP-N1 so that it is in frame with the EGFP coding sequences, with no intervening in-frame stop codons. The inserted gene should include the initiating ATG codon. The recombinant EGFP vector can be transfected into mammalian cells using any standard transfection method. If required, stable transformants can be selected using G418 . pEGFP-N1 can also be used simply to express EGFP in a cell line of interest (e.g., as a transfection marker).
质粒图谱：

ORIGIN

1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg

61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt

121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca

181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc

241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta

301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac

361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg

421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg

481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt

541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta

601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg

661 gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc

721 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc

781 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg

841 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc

901 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc

961 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag

1021 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac

1081 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg

1141 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac

1201 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg

1261 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag

1321 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg

1381 gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat accacatttg

1441 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa

1501 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca

1561 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt

1621 ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc

1681 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc

1741 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag

1801 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc

1861 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa

1921 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg

1981 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt

2041 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc

2101 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa

2161 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat

2221 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt

2281 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt

2341 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc

2401 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc

2461 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg

2521 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc

2581 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa

2641 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg

2701 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc

2761 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca

2821 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc

2881 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca

2941 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat

3001 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca

3061 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg

3121 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc

3181 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct

3241 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct

3301 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac

3361 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc

3421 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag

3481 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg

3541 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg

3601 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa

3661 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt

3721 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg

3781 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca

3841 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt

3901 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga

3961 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca

4021 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac

4081 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg

4141 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag

4201 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac

4261 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt

4321 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg

4381 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc

4441 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc

4501 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc

4561 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa

4621 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt

4681 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cat

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