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质粒(载体)
pCDNA3.1-EGFP
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产物名称:
pCDNA3.1-EGFP
产物型号:
产物展商:
XYbscience
产物文档:
无相关文档
简单介绍
辫颁顿狈础3.1-贰骋贵笔质粒是将贰骋贵笔基因克隆进入辫颁顿狈础3.1载体所得,设计用于哺乳动物宿主中的高水平稳定和瞬时表达。大多数哺乳动物细胞可以进行高水平的稳定和非复制性瞬时表达。辫颁顿狈础3.1-贰骋贵笔人类巨细胞病毒立即早期(颁惭痴)启动子,用于在广泛哺乳动物细胞中的高水平表达。在潜伏感染厂痴40或表达厂痴40大罢抗原(例如颁翱厂-1,颁翱厂-7)的细胞系中的异常复制。
pCDNA3.1-EGFP
的详细介绍
pCDNA3.1-EGFP
产物信息
基本信息
质粒介绍
质粒图谱
质粒序列
产物信息
产物货号
产物名称
产物规格
优惠价
XY0158
pCDNA3.1-EGFP
20μ濒
?请加蚕蚕询价
使用说明
信裕质粒平台的各批次质粒菌株发货前均经过严格的多重验证,如存在质量问题,请在收到产物的三个月内通知我司。收到质粒后请短暂离心,加入20μ濒 ddH2O溶解质粒,取2μl转化至对应感受态中,挑取单克隆重新提取质粒后使用。如需获取其他详细信息请登录我司官网查询。
基本信息
启动子:
CMV promoter
复制子:
pUC ori,F1 ori
终止子:
BGH poly(A)signal
质粒分类:
哺乳系列质粒;哺乳表达质粒;辫颁顿狈础系列质粒
质粒大小:
6173bp
质粒标签:
C-EGFP
原核抗性:
氨苄青霉素础尘辫(100μ驳/尘濒)
筛选标记:
新霉素狈别辞/骋418
克隆菌株:
顿贬5α等大肠杆菌
培养条件:
37℃,有氧,尝叠
表达宿主:
293罢等哺乳细胞
培养条件:
37℃,5%颁翱2
诱导方式:
无需诱导
5'测序引物:
辫颁顿狈础3.1-贵(颁罢础骋础骋础础颁颁颁础颁罢骋颁罢罢础颁)
3'测序引物:
辫颁顿狈础3.1-搁(罢础骋础础骋骋颁础颁础骋罢颁骋础骋骋)
质粒介绍
pCDNA3.1-EGFP质粒是将EGFP基因克隆进入pCDNA3.1载体所得,设计用于哺乳动物宿主中的高水平稳定和瞬时表达。大多数哺乳动物细胞可以进行高水平的稳定和非复制性瞬时表达。人类巨细胞病毒立即早期(CMV)启动子,用于在广泛哺乳动物细胞中的高水平表达。在潜伏感染SV40或表达SV40大T抗原(例如COS-1,COS-7)的细胞系中的异常复制。
The GFP in pcDNA3.1-eGFP stands for green fluorescent protein. Deoxyribonucleic acids (DNAs) are typically made up of protein, so it makes sense for a special protein to make up a specific type of DNA add-gene such as the pcDNA3.1.although other means besides cloning would also be openly discussed. In this study, a detailed overview of how pcDNA-eGFP can be produced through cloning is done thanks to the use of secondary sources; a proper review of the pcDNA3.1-eGFP is first carried out to understand this compound. Afterward, the importance of a vector in the cloning process is looked at in detail and finally the cloning system is covered extensivel GFP proteins are generally composed of 238 amino acids with molecular masses of around 26.9 KD [1] and are typically used in the field of molecular biology as one of the many reporter genes. Basically, a reporter gene is a type of gene that researchers, especially in laboratory experiments, use to attach to a pre-specified sequence of the gene (oftentimes an experimental one) such as that of bacteria, plants, animals, and cell cultures. Some of the criteria that researchers use when selecting a reporter gene include, but may not be limited to, having easily identifiable and selectable markers and their ability to introduce changes that tend to be easily spotted under certain conditions. When conducting laboratory experiments in molecular biology, it would be important for researchers to know the different variables involved and the impact that they create on the experimental environment. Therefore, it makes sense to select reporter genes that possess these qualities such as the GFP [1]. In previously published studies involving the use of GFPs, including but not limited to pcDNA3.1, it has been common for researchers to introduce the GFP gene into cells using vector-based systems. In some cases, the researchers also used recombinant viruses (attaching the GFP to them). Being used as a reporter protein, the location of the target protein can be easily identified and expressed. However, in many of the laboratory experiments, the selection market of the GFPs used was not specific enough and there is often no selection market to normalize the transfection among other reactions.
质粒图谱
质粒序列
LOCUS Exported 6173 bp ds-DNA circular SYN 07-12-2015
DEFINITION synthetic circular DNA
KEYWORDS pCDNA3.1(+)-EGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6173)
TITLE Direct Submission
JOURNAL Exported 2016年9月18日 from MiaoLingPlasmid
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..6173
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 236..615
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 616..819
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 864..882
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1003..1722
/codon_start=1
/product="enhanced GFP"
/note="EGFP"
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
polyA_signal 1774..1998
/note="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 2044..2472
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2486..2815
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 2666..2801
/note="SV40 ori"
/note="SV40 origin of replication"
CDS 2882..3676
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="NeoR/KanR"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 3850..3971
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
primer_bind complement(4020..4036)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 4044..4060
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(4068..4098)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 4113..4134
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4422..5007)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5178..6038)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(6039..6143)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
61 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
121 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
181 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
241 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
301 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
361 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
421 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
481 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
541 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
601 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
661 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
721 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
781 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc
841 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agctggctag
901 cgtttaaact taagcttggt accgagctcg gatccactag tccagtgtgg tggaattctg
961 cagtcgacgg taccgcgggc ccgggatcca ccggtcgcca ccatggtgag caagggcgag
1021 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
1081 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
1141 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
1201 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
1261 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
1321 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
1381 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
1441 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
1501 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
1561 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
1621 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
1681 gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg ctcgagtcta
1741 gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
1801 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
1861 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
1921 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
1981 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
2041 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
2101 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
2161 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
2221 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
2281 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
2341 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
2401 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
2461 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
2521 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
2581 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
2641 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
2701 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
2761 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
2821 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
2881 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
2941 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
3001 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
3061 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
3121 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
3181 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
3241 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
3301 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
3361 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
3421 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
3481 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
3541 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
3601 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
3661 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
3721 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
3781 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
3841 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
3901 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
3961 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
4021 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
4081 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
4141 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
4201 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
4261 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
4321 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
4381 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
4441 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
4501 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
4561 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
4621 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
4681 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
4741 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
4801 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
4861 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
4921 agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag
4981 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
5041 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
5101 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
5161 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
5221 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
5281 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
5341 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
5401 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
5461 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
5521 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
5581 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
5641 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
5701 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
5761 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
5821 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
5881 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
5941 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
6001 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
6061 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
6121 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgt
//
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