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pEGFP-C1质粒

货号：kl-zl-0659-01
培养条件：37℃，5%CO2

货号	产品名称	规格	库存	价格	数量	购买
kl-zl-0659-01	pEGFP-C1质粒	NA		立即咨询	+ -

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pEGFP-C1质粒货号：kl-zl-0659 规格：20ul
质粒描述：

启动子： CMV promoter

复制子： F1 ori,pUC ori，SV40 ori

终止子： SV40 poly(A) signal

质粒分类：哺乳系列质粒；哺乳荧光质粒；哺乳绿色质粒

质粒大小： 4731bp

质粒标签： C-EGFP

原核抗性：卡纳霉素Kan（50μg/ml）

筛选标记：新霉素Neo/G418

克隆菌株： DH5α等大肠杆菌

培养条件： 37℃，有氧 LB

表达宿主： 293T等哺乳细胞

培养条件： 37℃，5%CO2

诱导方式：无须诱导，瞬时表达

5'测序引物： pEGFP-C-5

3'测序引物： pEGFP-C-3

质粒简介：
pEGFP-C1编码野生型GFP的红移型变体，其已经针对更明亮的荧光和哺乳动物细胞中更高的表达进行了优化。（激发最大= 488nm;发射最大值= 507nm）pEGFP-C1编码GFPmut1变体，其含有Phe-64对Leu和Ser-65至Thr的双氨基酸取代pEGFPC1中的MCS在EGFP编码序列和SV40 polyA之间克隆到MCS中的基因如果与EGFP的阅读框处于相同的阅读框并且不存在中间终止密码子，则将其表达为EGFP的C末端的融合物。 EGFP基因下游的SV40聚腺苷酸信号直接适当处理EGFP mRNA的3'末端。载体骨架还含有用于在表达SV40 T抗原的哺乳动物细胞中复制的SV40来源。由SV40早期启动子，Tn5的新霉素/卡那霉素抗性基因和来自单纯疱疹病毒胸苷激酶（HSV TK）基因的多聚腺苷酸化信号组成的新霉素抗性盒（Neor）允许使用G418选择稳定转染的真核细胞。该盒上游的细菌启动子在大肠杆菌中表达卡那霉素抗性。 pEGFP-C1backbone还提供了用于在大肠杆菌中繁殖的pUC复制起点和用于单链DNA生产的f1来源。
pEGFP-C1encodes a red-shifted variant of wild-type GFP (1–3) which has been optimized for brighter fluorescence and higher expression in mammalian cells. (Excitation maximum = 488 nm;emission maximum = 507 nm.) pEGFP-C1encodes the GFPmut1 variant (4) 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 (5). Sequences flanking EGFP have been converted to a Kozak consensus translation initiation site (6) to further increase the translation efficiency in eukaryotic cells. The MCS in pEGFPC1is between the EGFP coding sequences and the SV40 poly A. Genes cloned into the MCS will be expressed as fusions to the C-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 (Neor), 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-C1backbone also provides a pUC origin of replication for propagation in E. coli and an f1 origin for single stranded DNA production.
Fusions to the C 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-C1 so that it is in frame with the EGFP coding sequences, with no intervening in-frame stop codons. 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-C1 can also be used simply to express EGFP in a cell line of interest (e.g., as a transfection marker).
Propagation in E. coli
• Suitable host strains： DH5α, HB101, and other general purpose strains. Single-stranded DNA production requires a host containing an F plasmid such as JM109 or XL1-Blue.
• Selectable marker： plasmid confers resistance to kanamycin (30 µg/ml) to E. coli hosts.
• E. coli replication origin： pUC
• Copy number： ≈500
• Plasmid incompatibility group： pMB1/ColE1
质粒图谱：

序列：

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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg

661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc

721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg

781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc

841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag

901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag

961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac

1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac

1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc

1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg

1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc

1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag

1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc

1381 gcgggcccgg gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta

1441 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg

1501 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat

1561 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc

1621 aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc

1681 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc

1741 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag

1801 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga

1861 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc

1921 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa

1981 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt

2041 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc

2101 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat

2161 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg

2221 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt

2281 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca

2341 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat

2401 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg

2461 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc

2521 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta

2581 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga

2641 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc

2701 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc

2761 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc

2821 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac

2881 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc

2941 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac

3001 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg

3061 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct

3121 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt

3181 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg

3241 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac

3301 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg

3361 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg

3421 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat

3481 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc

3541 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg

3601 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata

3661 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc

3721 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt

3781 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac

3841 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta

3901 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc

3961 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa

4021 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca

4081 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta

4141 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc

4201 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca

4261 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta

4321 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag

4381 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt

4441 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc

4501 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac

4561 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac

4621 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc

4681 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca t

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