| 质粒名称: | pEG BacMam |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 160451 |
| 存储实验室: | Eric Gouaux |
|
载体骨架基本信息 |
|
| 载体名称: | pFBDM |
| 载体出品公司: | https://www.nature.com/articles/nbt1036 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | 我们化学合成了源自先前描述的BacMam载体pVLAD的遗传元素（Dukkipati，A.，Park，H.H.，Waghray，D.，Fischer，S.&Garcia，K.C.BacMam系统，用于结构研究中重组可溶性糖蛋白和膜糖蛋白的高水平表达。蛋白质实验纯化62，160–170（2008））；用于有效RNA剪接和mRNA处理的CMV启动子和合成内含子，以及用于有效mRNA处理、稳定性和输出的WPRE基序。 |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 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: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | CMV |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pEG BacMam was a gift from Eric Gouaux (Addgene plasmid # 160451 ; http://n2t.net/addgene:160451 ; RRID:Addgene_160451) |
| 参考文献部分: | Screening and large-scale expression of membrane proteins in mammalian cells for structural studies. Goehring A, Lee CH, Wang KH, Michel JC, Claxton DP, Baconguis I, Althoff T, Fischer S, Garcia KC, Gouaux E. Nat Protoc. 2014 Nov;9(11):2574-85. doi: 10.1038/nprot.2014.173. Epub 2014 Oct 9. 10.1038/nprot.2014.173 PubMed 25299155 |
买家导航
pEGBacMam载体基本信息
pEGBacMam载体图谱质粒图谱和多克隆位点信息
pEGBacMam载体简介
pEGBacMam载体序列
LOCUS pEG BacMam 6893 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION vector optimized for use in screening assays, as well as for
efficient production of baculovirus and robust expression of the
target protein.
ACCESSION .
VERSION .
KEYWORDS pEG BacMam
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6893)
AUTHORS Goehring A, Lee CH, Wang KH, Michel JC, Claxton DP, Baconguis I,
Althoff T, Fischer S, Garcia KC, Gouaux E
TITLE Screening and large-scale expression of membrane proteins in
mammalian cells for structural studies.
JOURNAL Nat Protoc. 2014 Nov;9(11):2574-85. doi: 10.1038/nprot.2014.173.
Epub 2014 Oct 9.
PUBMED 25299155
REFERENCE 2 (bases 1 to 6893)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6893
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 3..206
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 157..177
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 203..227
/label=LNCX
/note="Human CMV promoter, forward primer"
intron 342..474
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
primer_bind 457..476
/label=pMT2-F
/note="Synthetic intron, forward primer"
misc_feature 607..1195
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(660..680)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(1078..1089)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 1328..1462
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1365..1384)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1419..1438
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
mobile_element 1491..1656
/mobile_element_type="transposon:Tn7"
/note="mini-Tn7 element (left end of the Tn7 transposon)"
rep_origin 1840..2294
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1927..1946)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2136..2157
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 2320..2424
/gene="bla"
/label=AmpR promoter
CDS 2425..3285
/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"
primer_bind complement(2643..2662)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3456..4044
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3945..3964
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4198..4215
/label=L4440
/note="L4440 vector, forward primer"
mobile_element 4349..4573
/mobile_element_type="transposon:Tn7"
/note="mini-Tn7 element (right end of the Tn7 transposon)"
CDS complement(4640..5173)
/codon_start=1
/gene="aacC1"
/product="gentamycin acetyltransferase"
/label=GmR
/note="confers resistance to gentamycin"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(5362..5390)
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
polyA_signal 5907..5955
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
primer_bind 5980..5999
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
promoter complement(6091..6200)
/label=p10 promoter
/note="baculovirus promoter for expression in insect cells"
promoter 6245..6336
/gene="polh from Autographa californica"
/label=polyhedrin promoter
/note="promoter for the baculovirus polyhedrin gene"
primer_bind 6261..6278
/label=Polyhedrin-F
/note="Polyhedrin promoter, forward primer"
enhancer 6516..2
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
ORIGIN
1 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
61 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
121 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
181 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcact agaagcttta
241 ttgcggtagt ttatcacagt taaattgcta acgcagtcag tgcttctgac acaacagtct
301 cgaacttaag ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa
361 gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt
421 tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc
481 actcccagtt caattacagc tcttaaggga tcccggtccg aagcgcgcgg aattcaaagg
541 cctacgtcga cgagctcact agtcgcggcc gctttcgaat ctagagcctg cagtctcgac
601 aagcttaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
661 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
721 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
781 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
841 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
901 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
961 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
1021 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
1081 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
1141 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgaa
1201 gcttgtcgag aagtactaga ggatcataat cagccatacc acatttgtag aggttttact
1261 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
1321 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
1381 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
1441 tgtatcttat catgtctgga tctgatcact gcttgagcct aggagatccg aaccagataa
1501 gtgaaatcta gttccaaact attttgtcat ttttaatttt cgtattagct tacgacgcta
1561 cacccagttc ccatctattt tgtcactctt ccctaaataa tccttaaaaa ctccatttcc
1621 acccctccca gttcccaact attttgtccg cccacagcgg ggcatttttc ttcctgttat
1681 gtttttaatc aaacatcctg ccaactccat gtgacaaacc gtcatcttcg gctacttttt
1741 ctctgtcaca gaatgaaaat ttttctgtca tctcttcgtt attaatgttt gtaattgact
1801 gaatatcaac gcttatttgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
1861 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
1921 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
1981 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
2041 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
2101 ttcgcccttt gacgttggag tccacgttct taatagtgga ctcttgttcc aaactggaac
2161 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
2221 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
2281 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
2341 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
2401 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
2461 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
2521 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
2581 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
2641 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
2701 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
2761 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
2821 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
2881 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
2941 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
3001 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
3061 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
3121 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
3181 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
3241 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
3301 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
3361 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
3421 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
3481 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
3541 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
3601 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
3661 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
3721 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
3781 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
3841 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
3901 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
3961 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
4021 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
4081 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
4141 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
4201 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
4261 tgcggtattt cacaccgcat agaccagccg cgtaacctgg caaaatcggt tacggttgag
4321 taataaatgg atgccctgcg taagcgggtg tgggcggaca ataaagtctt aaactgaaca
4381 aaatagatct aaactatgac aataaagtct taaactagac agaatagttg taaactgaaa
4441 tcagtccagt tatgctgtga aaaagcatac tggacttttg ttatggctaa agcaaactct
4501 tcattttctg aagtgcaaat tgcccgtcgt attaaagagg ggcgtggcca agggcatggt
4561 aaagactata ttcgcggcgt tgtgacaatt taccgaacaa ctccgcggcc gggaagccga
4621 tctcggcttg aacgaattgt taggtggcgg tacttgggtc gatatcaaag tgcatcactt
4681 cttcccgtat gcccaacttt gtatagagag ccactgcggg atcgtcaccg taatctgctt
4741 gcacgtagat cacataagca ccaagcgcgt tggcctcatg cttgaggaga ttgatgagcg
4801 cggtggcaat gccctgcctc cggtgctcgc cggagactgc gagatcatag atatagatct
4861 cactacgcgg ctgctcaaac ttgggcagaa cgtaagccgc gagagcgcca acaaccgctt
4921 cttggtcgaa ggcagcaagc gcgatgaatg tcttactacg gagcaagttc ccgaggtaat
4981 cggagtccgg ctgatgttgg gagtaggtgg ctacgtctcc gaactcacga ccgaaaagat
5041 caagagcagc ccgcatggat ttgacttggt cagggccgag cctacatgtg cgaatgatgc
5101 ccatacttga gccacctaac tttgttttag ggcgactgcc ctgctgcgta acatcgttgc
5161 tgctgcgtaa catcgttgct gctccataac atcaaacatc gacccacggc gtaacgcgct
5221 tgctgcttgg atgcccgagg catagactgt acaaaaaaac agtcataaca agccatgaaa
5281 accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgat
5341 cgcatacgct acttgcatta cagtttacga accgaacagg cttatgtcaa ctgggttcgt
5401 gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct tgggcagcag cgaagtcgag
5461 gcatttctgt cctggctggc gaacgagcgc aaggtttcgg tctccacgca tcgtcaggca
5521 ttggcggcct tgctgttctt ctacggcaag gtgctgtgca cggatctgcc cttgcttcag
5581 gagatcggta gacctcggcc gtcgcggcgc ttgccggtgg tgctgacccc ggatgaagtg
5641 gttcgcatcc tcggttttct ggaaggcgag catcgtttgt tcgcccagga ctctagctat
5701 agttctagtg gttggctaca gctttgttta aacaaagctg gctatggcag ggcttgccgc
5761 cccgacgttg gctgcgagcc ctgggccttc acccgaactt gggggttggg gtggggaaaa
5821 ggaagaaacg cgggcgtatt ggtcccaatg gggtctcggt ggggtatcga cagagtgcca
5881 gccctgggac cgaaccccgc gtttatgaac aaacgaccca acacccgtgc gttttattct
5941 gtctttttat tgccgtcata gcgcgggttc cttccggtat tgtctccttc cgtgtttcag
6001 ttagcctccc ccatctcccg gtaccgcatg ctatgcatca gctgctagca ccatggctcg
6061 agatcccggg tgatcaagtc ttcgtcgagt gattgtaaat aaaatgtaat ttacagtata
6121 gtattttaat taatatacaa atgatttgat aataattctt atttaactat aatatattgt
6181 gttgggttga attaaaggtc cgtatactag tatcgattcg cgacctactc cggaatatta
6241 atagatcatg gagataatta aaatgataac catctcgcaa ataaataagt attttactgt
6301 tttcgtaaca gttttgtaat aaaaaaacct ataaatattc cggattattc ataccgtccc
6361 accatcgggc gcggatcctc aatattggcc attagccata ttattcattg gttatatagc
6421 ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata atatgtacat
6481 ttatattggc tcatgtccaa tatgaccgcc atgttggcat tgattattga ctagttatta
6541 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
6601 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
6661 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
6721 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc
6781 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
6841 acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta cca
//
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