| 质粒名称: | eGFP-luciferase-C1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 163523 |
| 存储实验室: | Thomas Weber |
|
载体骨架基本信息 |
|
| 载体名称: | eGFP-C1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4731bp |
| 完整质粒大小: | 6366bp |
| 载体类型: | 哺乳细胞表达, Bacterial Expression, Luciferase |
| 载体抗性: | Kanamycin |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Firefly luciferase |
| 物种来源: | Fire Fly |
| 插入大小: | 1688bp |
| 标签/融合蛋白: | eGFP (N terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | Xho1 (unknown if destroyed) |
| 3’克隆位点: | BamH1 (unknown if destroyed) |
| 5’测序引物 | CATGGTCCTGCTGGAGTTCG |
|
文献引用方法 |
|
| 材料和方法部分: |
eGFP-luciferase-C1 was a gift from Thomas Weber (Addgene plasmid # 163523 ; http://n2t.net/addgene:163523 ; RRID:Addgene_163523)
|
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS eGFP-luciferase-C1 6366 bp DNA circular SYN 25-AUG-2024
DEFINITION Expresses both eGFP and firefly luciferase in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6366)
TITLE Expression plasmid for production of murine IFN gamma
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6366)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
FEATURES Location/Qualifiers
source 1..6366
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 61..364
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 519..539
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 607..616
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 613..1329
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 613..1329
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(658..679)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(919..938)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1266..1287
/label=EGFP-C
/note="EGFP, forward primer"
CDS 1372..3024
/codon_start=1
/gene="luc+"
/product="firefly luciferase"
/label=luciferase
/note="enhanced luc+ version of the luciferase gene"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
primer_bind complement(1438..1456)
/label=LucNrev
/note="Firefly luciferase, reverse primer"
polyA_signal 3154..3275
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3191..3210)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3245..3264
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(3282..3737)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3419..3440)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3631..3650
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3764..3868
/gene="bla"
/label=AmpR promoter
promoter 3870..4227
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 3870..4061
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(3893..3913)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 4078..4213
/label=SV40 ori
/note="SV40 origin of replication"
CDS 4262..5056
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
CDS 4262..5056
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(4316..4335)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4926..4945
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(5244..5263)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5288..5335
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5664..6252
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6153..6172
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
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 tatggaagac
1381 gccaaaaaca taaagaaagg cccggcgcca ttctatccgc tggaagatgg aaccgctgga
1441 gagcaactgc ataaggctat gaagagatac gccctggttc ctggaacaat tgcttttaca
1501 gatgcacata tcgaggtgga catcacttac gctgagtact tcgaaatgtc cgttcggttg
1561 gcagaagcta tgaaacgata tgggctgaat acaaatcaca gaatcgtcgt atgcagtgaa
1621 aactctcttc aattctttat gccggtgttg ggcgcgttat ttatcggagt tgcagttgcg
1681 cccgcgaacg acatttataa tgaacgtgaa ttgctcaaca gtatgggcat ttcgcagcct
1741 accgtggtgt tcgtttccaa aaaggggttg caaaaaattt tgaacgtgca aaaaaagctc
1801 ccaatcatcc aaaaaattat tatcatggat tctaaaacgg attaccaggg atttcagtcg
1861 atgtacacgt tcgtcacatc tcatctacct cccggtttta atgaatacga ttttgtgcca
1921 gagtccttcg atagggacaa gacaattgca ctgatcatga actcctctgg atctactggt
1981 ctgcctaaag gtgtcgctct gcctcataga actgcctgcg tgagattctc gcatgccaga
2041 gatcctattt ttggcaatca aatcattccg gatactgcga ttttaagtgt tgttccattc
2101 catcacggtt ttggaatgtt tactacactc ggatatttga tatgtggatt tcgagtcgtc
2161 ttaatgtata gatttgaaga agagctgttt ctgaggagcc ttcaggatta caagattcaa
2221 agtgcgctgc tggtgccaac cctattctcc ttcttcgcca aaagcactct gattgacaaa
2281 tacgatttat ctaatttaca cgaaattgct tctggtggcg ctcccctctc taaggaagtc
2341 ggggaagcgg ttgccaagag gttccatctg ccaggtatca ggcaaggata tgggctcact
2401 gagactacat cagctattct gattacaccc gagggggatg ataaaccggg cgcggtcggt
2461 aaagttgttc cattttttga agcgaaggtt gtggatctgg ataccgggaa aacgctgggc
2521 gttaatcaaa gaggcgaact gtgtgtgaga ggtcctatga ttatgtccgg ttatgtaaac
2581 aatccggaag cgaccaacgc cttgattgac aaggatggat ggctacattc tggagacata
2641 gcttactggg acgaagacga acacttcttc atcgttgacc gcctgaagtc tctgattaag
2701 tacaaaggct atcaggtggc tcccgctgaa ttggaatcca tcttgctcca acaccccaac
2761 atcttcgacg caggtgtcgc aggtcttccc gacgatgacg ccggtgaact tcccgccgcc
2821 gttgttgttt tggagcacgg aaagacgatg acggaaaaag agatcgtgga ttacgtcgcc
2881 agtcaagtaa caaccgcgaa aaagttgcgc ggaggagttg tgtttgtgga cgaagtaccg
2941 aaaggtctta ccggaaaact cgacgcaaga aaaatcagag agatcctcat aaaggccaag
3001 aagggcggaa agatcgccgt gtaaggatcc accggatcta gataactgat cataatcagc
3061 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
3121 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
3181 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
3241 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat
3301 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
3361 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
3421 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
3481 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
3541 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
3601 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
3661 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
3721 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
3781 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
3841 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
3901 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
3961 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
4021 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
4081 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
4141 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
4201 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
4261 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
4321 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
4381 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
4441 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
4501 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
4561 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
4621 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
4681 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
4741 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
4801 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
4861 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
4921 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
4981 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
5041 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
5101 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
5161 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
5221 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
5281 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
5341 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
5401 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
5461 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
5521 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
5581 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
5641 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
5701 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
5761 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
5821 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
5881 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
5941 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
6001 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
6061 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
6121 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
6181 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
6241 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
6301 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
6361 atgcat
//
载体图谱质粒图谱pdf版和相关资料下载
载体质粒应用举例
- 上一篇：mAmetrine-N1
- 下一篇：没有了
