| 质粒名称: | pEZT-BM |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 74099 |
| 存储实验室: | Ryan Hibbs |
|
载体骨架基本信息 |
|
| 载体名称: | Modified pFastBacDual |
| 载体出品公司: | Thermo Fisher Scientific |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | 从pVLAD6载体中移除多面体启动子并替换为含有CMV的盒体（Dukkipati等人，蛋白质表达纯化，2008年）。多个克隆站点已替换为自定义。合成，密码子优化的EGFP插入p10启动子后。参见Morales Perez等人，结构，2016年，了解向量的全部细节。 |
| 载体类型: | 哺乳动物表达；杆状病毒 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
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, p10 |
| 标签/融合蛋白: | p10启动子后的EGFP |
| 克隆方法: | Ligation Independent Cloning |
| 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: | pEZT-BM was a gift from Ryan Hibbs (Addgene plasmid # 74099 ; http://n2t.net/addgene:74099 ; RRID:Addgene_74099) |
| 参考文献部分: | Manipulation of Subunit Stoichiometry in Heteromeric Membrane Proteins. Morales-Perez CL, Noviello CM, Hibbs RE. Structure. 2016 Mar 24. pii: S0969-2126(16)00082-4. doi: 10.1016/j.str.2016.03.004. 10.1016/j.str.2016.03.004 PubMed 27041595 |
买家导航
pEZTBM载体基本信息
pEZTBM载体图谱质粒图谱和多克隆位点信息
pEZTBM载体简介
pEZTBM载体序列
LOCUS pEZT-BM 7427 bp ds-DNA circular SYN 22-APR-2019
DEFINITION Multi-purpose rapid titer bacmam expression vector.
ACCESSION .
VERSION .
KEYWORDS pEZT-BM
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7427)
AUTHORS Morales-Perez CL, Noviello CM, Hibbs RE
TITLE Manipulation of Subunit Stoichiometry in Heteromeric Membrane
Proteins.
JOURNAL Structure. 2016 Mar 24. pii: S0969-2126(16)00082-4. doi:
10.1016/j.str.2016.03.004.
PUBMED 27041595
REFERENCE 2 (bases 1 to 7427)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 22, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7427
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 102..556
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(189..208)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 398..419
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 582..686
/gene="bla"
/label=AmpR promoter
CDS 687..1547
/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(905..924)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1718..2306
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2207..2226
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2460..2477
/label=L4440
/note="L4440 vector, forward primer"
mobile_element 2611..2835
/mobile_element_type="transposon:Tn7"
/note="mini-Tn7 element (right end of the Tn7 transposon)"
CDS complement(2902..3435)
/codon_start=1
/gene="aacC1"
/product="gentamycin acetyltransferase"
/label=GmR
/note="confers resistance to gentamycin"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(3624..3652)
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
polyA_signal 4160..4208
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
primer_bind 4233..4252
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
CDS complement(4293..5006)
/codon_start=1
/product="enhanced GFP with monomerizing A206K mutation
(Zacharias et al., 2002)"
/label=mEGFP
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(4335..4356)
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 4684..4703
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4943..4964
/label=EGFP-N
/note="EGFP, reverse primer"
promoter complement(5031..5140)
/label=p10 promoter
/note="baculovirus promoter for expression in insect cells"
enhancer 5325..5704
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 5705..5908
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 5859..5879
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 5905..5929
/label=LNCX
/note="Human CMV promoter, forward primer"
intron 6044..6176
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
primer_bind 6159..6178
/label=pMT2-F
/note="Synthetic intron, forward primer"
promoter 6221..6239
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 6287..6875
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(6340..6360)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(6758..6769)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 7017..7151
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(7054..7073)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 7108..7127
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
mobile_element 7180..7345
/mobile_element_type="transposon:Tn7"
/note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
1 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga
61 ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
241 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
361 ttttcgccct ttgacgttgg agtccacgtt cttaatagtg gactcttgtt ccaaactgga
421 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
481 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
541 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
601 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
661 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
721 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
781 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
841 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
901 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
961 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
1021 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
1081 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
1141 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
1201 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
1261 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
1321 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
1381 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
1441 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
1501 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
1561 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
1621 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
1681 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
1741 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
1801 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
1861 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
1921 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
1981 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
2041 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
2101 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
2161 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
2221 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
2281 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
2341 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
2401 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
2461 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
2521 tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct ggcaaaatcg gttacggttg
2581 agtaataaat ggatgccctg cgtaagcggg tgtgggcgga caataaagtc ttaaactgaa
2641 caaaatagat ctaaactatg acaataaagt cttaaactag acagaatagt tgtaaactga
2701 aatcagtcca gttatgctgt gaaaaagcat actggacttt tgttatggct aaagcaaact
2761 cttcattttc tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggc caagggcatg
2821 gtaaagacta tattcgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc
2881 gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac
2941 ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc
3001 ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag
3061 cgcggtggca atgccctgcc tccggtgctc gccggagact gcgagatcat agatatagat
3121 ctcactacgc ggctgctcaa acttgggcag aacgtaagcc gcgagagcgc caacaaccgc
3181 ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta
3241 atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag
3301 atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat
3361 gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt
3421 gctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg
3481 cttgctgctt ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga
3541 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg
3601 agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttc
3661 gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg
3721 aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg
3781 cattggcggc cttgctgttc ttctacggca aggtgctgtg cacggatctg cccttgcttc
3841 aggagatcgg tagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag
3901 tggttcgcat cctcggtttt ctggaaggcg agcatcgttt gttcgcccag gactctagct
3961 atagttctag tggttggcta cgtacccgta gtggctatgg cagggcttgc cgccccgacg
4021 ttggctgcga gccctgggcc ttcacccgaa cttgggggtt ggggtgggga aaaggaagaa
4081 acgcgggcgt attggtccca atggggtctc ggtggggtat cgacagagtg ccagccctgg
4141 gaccgaaccc cgcgtttatg aacaaacgac ccaacacccg tgcgttttat tctgtctttt
4201 tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt cagttagcct
4261 cccccatctc ccggttccgc atgctatgca tccttgtaca gctcgtccat gccgagagtg
4321 atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct
4381 ttgctcagct tggactgggt gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg
4441 ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg
4501 tggcggatct tgaagttcac cttgatgccg ttcttctgct tgtcggccat gatatagacg
4561 ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag
4621 tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg
4681 cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg
4741 ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac
4801 tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg
4861 gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac
4921 acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg
4981 gtgaacagct cctcgccctt gctcaccggg tgatcaagtc ttcgtcgagt gattgtaaat
5041 aaaatgtaat ttacagtata gtattttaat taatatacaa atgatttgat aataattctt
5101 atttaactat aatatattgt gttgggttga attaaaggtc cgtatactcc ggaatattaa
5161 taggatcccg gtccgagcgc gcggaattca atattggcca ttagccatat tattcattgg
5221 ttatatagca taaatcaata ttggctattg gccattgcat acgttgtatc tatatcataa
5281 tatgtacatt tatattggct catgtccaat atgaccgcca tgttggcatt gattattgac
5341 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
5401 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
5461 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
5521 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
5581 aagtccgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
5641 catgacctta cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
5701 catggtgatg cggttttggc agtacaccaa tgggcgtgga tagcggtttg actcacgggg
5761 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
5821 ggactttcca aaatgtcgta ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt
5881 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatca ctagaagctt
5941 tattgcggta gtttatcaca gttaaattgc taacgcagtc agtgcttctg acacaacagt
6001 ctcgaactta agctgcagaa gttggtcgtg aggcactggg caggtaagta tcaaggttac
6061 aagacaggtt taaggagacc aatagaaact gggcttgtcg agacagagaa gactcttgcg
6121 tttctgatag gcacctattg gtcttactga catccacttt gcctttctct ccacaggtgt
6181 ccactcccag ttcaattaca gctcttaagg ctagagtact taatacgact cactataggc
6241 tagcctcgag gcggccgcga ctgcagggat ccgaggtacc aagcttaatc aacctctgga
6301 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
6361 tgaatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
6421 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
6481 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
6541 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
6601 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
6661 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
6721 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
6781 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
6841 gacgagtcgg atctcccttt gggccgcctc cccgcctgca gtctgacaag cttgtcgaga
6901 agtactagag gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
6961 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
7021 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
7081 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
7141 atgtctggat ctgatcactg cttgagccta ggagatccga accagataag tgaaatctag
7201 ttccaaacta ttttgtcatt tttaattttc gtattagctt acgacgctac acccagttcc
7261 catctatttt gtcactcttc cctaaataat ccttaaaaac tccatttcca cccctcccag
7321 ttcccaacta ttttgtccgc ccacagcggg gcatttttct tcctgttatg tttttaatca
7381 aacatcctgc caactccatg tgacaaaccg tcatcttcgg ctacttt
//
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