| 质粒名称: | EGFR-mEGFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 203774 |
| 存储实验室: | Thorsten Wohland |
|
载体骨架基本信息 |
|
| 载体名称: | pRK5 |
| 载体出品公司: | Clontech |
| 空载体大小: | 2925 bp |
| 完整质粒大小: | 7314 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | iRFP713 |
| 物种来源: | 光合细菌 Rhodopseudomonas palustris |
| 插入大小: | 951 bp |
| 基因突变: | T2A, S13L, A92T, V104T, V114I, E161K, Y193K, F198Y, D202T, I203V, Y258F, A283V, K288T, N290Y relative to RpBphP2 |
| GenBank编号 | JN247409 |
| 启动子: | CMV |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | AgeI(未破坏) |
| 3’克隆位点: | NotI (未破坏) |
| 5’测序引物 | CMV-F |
| 3’测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | |
| 参考文献部分: | |
买家导航
EGFR-mEGFP载体质粒基本信息
EGFR-mEGFP质粒图谱载体图谱和 EGFR-mEGFP载体序列质粒序列多克隆位点信息
EGFR-mEGFP质粒载体简介
EGFR-mEGFP质粒序列载体序列
LOCUS EGFR-mEGFP 7314 bp DNA circular SYN 14-APR-2025
DEFINITION Number and brightness analysis of EGFR oligomerization.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7314)
AUTHORS Gupta A, Lu D, Balasubramanian H, Chi Z, Wohland T
TITLE Heptanol-mediated phase separation determines phase preference of
molecules in live cell membranes.
JOURNAL J Lipid Res. 2022 Jun;63(6):100220. doi: 10.1016/j.jlr.2022.100220.
Epub 2022 Apr 28.
PUBMED 35490741
REFERENCE 2 (bases 1 to 7314)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Lipid
Res."; date: "2022-06"; volume: "63(6):100220. doi"; pages: "
10.1016/j.jlr.2022.100220. Epub 2022 Apr 28"
COMMENT Sequence Label: EGFR-mEGFP
FEATURES Location/Qualifiers
source 1..7314
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 1..21
/gene="mutant version of attB"
/label=attB4
/bound_moiety="BP Clonase(TM)"
/note="core recombination site for the Gateway(R) BP
reaction"
enhancer 82..385
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 386..589
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 540..560
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS join(623..694,695..2557,2558..2626,2627..4252)
/codon_start=1
/product="precursor of human epidermal growth factor
receptor"
/label=EGFR
/translation="MRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFE
DHFLSLQRMFNNCEVVLGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENL
QIIRGNMYYENSYALAVLSNYDANKTGLKELPMRNLQEILHGAVRFSNNPALCNVESIQ
WRDIVSSDFLSNMSMDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGR
CRGKSPSDCCHNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNP
EGKYSFGATCVKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGI
GIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVK
EITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISD
GDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWG
PEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGP
DNCIQCAHYIDGPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEG
CPTNGPKIPSIATGMVGALLLLLVVALGIGLFMRRRHIVRKRTLRRLLQERELVEPLTP
SGEAPNQALLRILKETEFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSP
KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ
YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA
EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG
ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL
PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA
CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN
PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS
LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
CDS join(4271..4273,4274..4276,4277..4987)
/codon_start=1
/product="enhanced GFP with monomerizing A206K mutation
(Zacharias et al., 2002)"
/label=mEGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(4316..4337)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(4577..4596)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4924..4945
/label=EGFP-C
/note="EGFP, forward primer"
protein_bind complement(5018..5042)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
polyA_signal 5071..5192
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5082..5101)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 5136..5155
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(5308..5325)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5479..6067)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5559..5578)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(6238..7029,7030..7098))
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(7099..7203)
/gene="bla"
/label=AmpR promoter
primer_bind 7271..7289
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 caactttgta tagaaaagtt gtagttatta atagtaatca attacggggt cattagttca
61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
361 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
421 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
481 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
541 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
601 ccgtcagatc accggtgcca ccatgcgacc ctccgggacg gccggggcag cgctcctggc
661 gctgctggct gcgctctgcc cggcgagtcg ggctctggag gaaaagaaag tttgccaagg
721 cacgagtaac aagctcacgc agttgggcac ttttgaagat cattttctca gcctccagag
781 gatgttcaat aactgtgagg tggtccttgg gaatttggaa attacctatg tgcagaggaa
841 ttatgatctt tccttcttaa agaccatcca ggaggtggct ggttatgtcc tcattgccct
901 caacacagtg gagcgaattc ctttggaaaa cctgcagatc atcagaggaa atatgtacta
961 cgaaaattcc tatgccttag cagtcttatc taactatgat gcaaataaaa ccggactgaa
1021 ggagctgccc atgagaaatt tacaggaaat cctgcatggc gccgtgcggt tcagcaacaa
1081 ccctgccctg tgcaacgtgg agagcatcca gtggcgggac atagtcagca gtgactttct
1141 cagcaacatg tcgatggact tccagaacca cctgggcagc tgccaaaagt gtgatccaag
1201 ctgtcccaat gggagctgct ggggtgcagg agaggagaac tgccagaaac tgaccaaaat
1261 catctgtgcc cagcagtgct ccgggcgctg ccgtggcaag tcccccagtg actgctgcca
1321 caaccagtgt gctgcaggct gcacaggccc ccgggagagc gactgcctgg tctgccgcaa
1381 attccgagac gaagccacgt gcaaggacac ctgcccccca ctcatgctct acaaccccac
1441 cacgtaccag atggatgtga accccgaggg caaatacagc tttggtgcca cctgcgtgaa
1501 gaagtgtccc cgtaattatg tggtgacaga tcacggctcg tgcgtccgag cctgtggggc
1561 cgacagctat gagatggagg aagacggcgt ccgcaagtgt aagaagtgcg aagggccttg
1621 ccgcaaagtg tgtaacggaa taggtattgg tgaatttaaa gactcactct ccataaatgc
1681 tacgaatatt aaacacttca aaaactgcac ctccatcagt ggcgatctcc acatcctgcc
1741 ggtggcattt aggggtgact ccttcacaca tactcctcct ctggatccac aggaactgga
1801 tattctgaaa accgtaaagg aaatcacagg gtttttgctg attcaggctt ggcctgaaaa
1861 caggacggac ctccatgcct ttgagaacct agaaatcata cgcggcagga ccaagcaaca
1921 tggtcagttt tctcttgcag tcgtcagcct gaacataaca tccttgggat tacgctccct
1981 caaggagata agtgatggag atgtgataat ttcaggaaac aaaaatttgt gctatgcaaa
2041 tacaataaac tggaaaaaac tgtttgggac ctccggtcag aaaaccaaaa ttataagcaa
2101 cagaggtgaa aacagctgca aggccacagg ccaggtctgc catgccttgt gctcccccga
2161 gggctgctgg ggcccggagc ccagggactg cgtctcttgc cggaatgtca gccgaggcag
2221 ggaatgcgtg gacaagtgca accttctgga gggtgagcca agggagtttg tggagaactc
2281 tgagtgcata cagtgccacc cagagtgcct gcctcaggcc atgaacatca cctgcacagg
2341 acggggacca gacaactgta tccagtgtgc ccactacatt gacggccccc actgcgtcaa
2401 gacctgcccg gcaggagtca tgggagaaaa caacaccctg gtctggaagt acgcagacgc
2461 cggccatgtg tgccacctgt gccatccaaa ctgcacctac ggatgcactg ggccaggtct
2521 tgaaggctgt ccaacgaatg ggcctaagat cccgtccatc gccactggga tggtgggggc
2581 cctcctcttg ctgctggtgg tggccctggg gatcggcctc ttcatgcgaa ggcgccacat
2641 cgttcggaag cgcacgctgc ggaggctgct gcaggagagg gagcttgtgg agcctcttac
2701 acccagtgga gaagctccca accaagctct cttgaggatc ttgaaggaaa ctgaattcaa
2761 aaagatcaaa gtgctgggct ccggtgcgtt cggcacggtg tataagggac tctggatccc
2821 agaaggtgag aaagttaaaa ttcccgtcgc tatcaaggaa ttaagagaag caacatctcc
2881 gaaagccaac aaggaaatcc tcgatgaagc ctacgtgatg gccagcgtgg acaaccccca
2941 cgtgtgccgc ctgctgggca tctgcctcac ctccaccgtg caactcatca cgcagctcat
3001 gcccttcggc tgcctcctgg actatgtccg ggaacacaaa gacaatattg gctcccagta
3061 cctgctcaac tggtgtgtgc agatcgcaaa gggcatgaac tacttggagg accgtcgctt
3121 ggtgcaccgc gacctggcag ccaggaacgt actggtgaaa acaccgcagc atgtcaagat
3181 cacagatttt gggctggcca aactgctggg tgcggaagag aaagaatacc atgcagaagg
3241 aggcaaagtg cctatcaagt ggatggcatt ggaatcaatt ttacacagaa tctataccca
3301 ccagagtgat gtctggagct acggggtgac cgtttgggag ttgatgacct ttggatccaa
3361 gccatatgac ggaatccctg ccagcgagat ctcctccatc ctggagaaag gagaacgcct
3421 ccctcagcca cccatatgta ccatcgatgt ctacatgatc atggtcaagt gctggatgat
3481 agacgcagat agtcgcccaa agttccgtga gttgatcatc gaattctcca aaatggcccg
3541 agacccccag cgctaccttg tcattcaggg ggatgaaaga atgcatttgc caagtcctac
3601 agactccaac ttctaccgtg ccctgatgga tgaagaagac atggacgacg tggtggatgc
3661 cgacgagtac ctcatcccac agcagggctt cttcagcagc ccctccacgt cacggactcc
3721 cctcctgagc tctctgagtg caaccagcaa caattccacc gtggcttgca ttgatagaaa
3781 tgggctgcaa agctgtccca tcaaggaaga cagcttcttg cagcgataca gctcagaccc
3841 cacaggcgcc ttgactgagg acagcataga cgacaccttc ctcccagtgc ctgaatacat
3901 aaaccagtcc gttcccaaaa ggcccgctgg ctctgtgcag aatcctgtct atcacaatca
3961 gcctctgaac cccgcgccca gcagagaccc acactaccag gacccccaca gcactgcagt
4021 gggcaacccc gagtatctca acactgtcca gcccacctgt gtcaacagca cattcgacag
4081 ccctgcccac tgggcccaga aaggcagcca ccaaattagc ctggacaacc ctgactacca
4141 gcaggacttc tttcccaagg aagccaagcc aaatggcatc tttaagggct ccacagctga
4201 aaatgcagaa tacctaaggg tcgcgccaca aagcagtgaa tttattggag caggaggtgg
4261 cgggactagt atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
4321 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
4381 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
4441 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
4501 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
4561 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
4621 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
4681 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
4741 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
4801 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
4861 cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga
4921 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
4981 gtacaagggt ggcggagggt ccgctagcta aaagcttacc cagctttctt gtacaaagtg
5041 gtgatggccg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
5101 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
5161 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
5221 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
5281 gtagcggccg cggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
5341 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
5401 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
5461 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
5521 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
5581 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
5641 ctttctctct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
5701 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
5761 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
5821 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
5881 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
5941 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
6001 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
6061 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
6121 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
6181 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
6241 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
6301 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
6361 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
6421 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
6481 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
6541 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
6601 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
6661 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
6721 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
6781 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
6841 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
6901 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
6961 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
7021 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
7081 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
7141 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
7201 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
7261 aacctataaa aataggcgta tcacgaggcc ctttcgtcgg cgcgccgcgg ccgc
//
