| 质粒名称: | pHLmMBP-10 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 72348 |
| 存储实验室: | Luca Jovine |
|
载体骨架基本信息 |
|
| 载体名称: | pHLmMBP-10 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 5931 bp |
| 载体修饰: | 人细胞色素c氧化酶亚基VIII（human cytochrome C oxidase subunit VIII） |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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 | -- |
| 标签/融合蛋白: | -- |
| 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: | pHLmMBP-10 was a gift from Luca Jovine (Addgene plasmid # 72348 ; http://n2t.net/addgene:72348 ; RRID:Addgene_72348) |
| 参考文献部分: | Easy mammalian expression and crystallography of maltose-binding protein-fused human proteins. Bokhove M, Sadat Al Hosseini H, Saito T, Dioguardi E, Gegenschatz-Schmid K, Nishimura K, Raj I, de Sanctis D, Han L, Jovine L. J Struct Biol. 2016 Feb 3. 194:1-7. pii: S1047-8477(16)30015-6. doi: 10.1016/j.jsb.2016.01.016. 10.1016/j.jsb.2016.01.016 PubMed 26850170 |
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 pHLmMBP-10 5931 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Mammalian expression of secreted N-terminally 8His-tagged mMBP TEV
cleavage site-linked fusion proteins with C-terminal
6His-tag/Strep-tag II/HA-tag.
ACCESSION .
VERSION .
KEYWORDS pHLmMBP-10
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5931)
AUTHORS Bokhove M, Sadat Al Hosseini H, Saito T, Dioguardi E,
Gegenschatz-Schmid K, Nishimura K, Raj I, de Sanctis D, Han L,
Jovine L
TITLE Easy mammalian expression and crystallography of maltose-binding
protein-fused human proteins.
JOURNAL J Struct Biol. 2016 Feb 3. 194:1-7. pii: S1047-8477(16)30015-6. doi:
10.1016/j.jsb.2016.01.016.
PUBMED 26850170
REFERENCE 2 (bases 1 to 5931)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5931
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 4..383
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 386..661
/label=chicken beta-actin promoter
intron 662..1670
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1594..1616
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
regulatory 1758..1767
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1857..1880
/codon_start=1
/product="8xHis affinity tag"
/label=8xHis
/translation="HHHHHHHH"
CDS 3015..3035
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
CDS 3066..3083
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 3111..3134
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS 3153..3179
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
primer_bind complement(3288..3307)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_site 3353..3408
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(3407..3426)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(3767..3783)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3767..3783)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3780..3802)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3791..3807
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(3815..3845)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3860..3881
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4010..4027)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4181..4769)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4261..4280)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4940..5800)
/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 5563..5582
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5801..5905)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
1681 acgtgctggt tgttgtgctg tctcatcatt ttggcaaaga attgcggccg tctcaggccg
1741 aattcgtcga caagcttgcc accatgggga tccttcccag ccctgggatg cctgcgctgc
1801 tctccctcgt gagccttctc tccgtgctgc tgatgggttg cgtagctgaa accggtcacc
1861 atcaccatca ccatcaccat aaaaccgaag aaggtaaact ggtaatctgg attaacggcg
1921 ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat accggaatta
1981 aagtcaccgt tgagcatccc gataaactgg aagagaaatt cccacaggtt gcggcaactg
2041 gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac gctcaatctg
2101 gcctgttggc tgaaatcacc ccggccgcag cgttccagga caagctgtat ccgtttacct
2161 gggatgccgt gcgctacaac ggcaagctga ttgcttaccc gatcgctgtt gaagcgttgt
2221 ccctgattta taacaaggat ctgctgccga acccgccaaa aacctgggaa gagatcccgg
2281 cgctggataa agaactgaaa gcgaaaggta agagcgcgct gatgttcaac ctgcaagaac
2341 cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag tatgcagccg
2401 gaaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg ggtctgacct
2461 tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac tccatcgcag
2521 aacatgcctt taatcatggc gaaacagcga tgaccatcaa cggcccgtgg gcatggtcca
2581 acatcgacac cagcgcagtg aattatggtg taacggtgct gccgaccttc aagggtcaac
2641 catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt ccgaacaaag
2701 agctggcaaa agagttcctc gaaaactacc tgctgactga tgaaggtctg gaagcggtta
2761 ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag ttggtgaaag
2821 atccacgtgt tgccgccaca atggaaaacg cccagaaagg tgaaatcatg cccaacatcc
2881 cgcagatgtc cgctttctgg tatgccgtgc gcactgcggt gattaacgcc gccagcggtc
2941 gtcagactgt cgatgcagcc ctggcagccg cgcagactaa tatcaccaag ggcggaggga
3001 gcggcggagg gagcgaaaac ctgtattttc agagcgcggc cgccgcgagc tcggctagcc
3061 tcgagcacca tcaccatcac cattgataaa cgcgtttaat taaccccggg tggagccacc
3121 cccagttcga gaagtgataa gatgatcaag gatatcccta cgacgtgccc gactacgcct
3181 gataatctag aactagtatc gcgataattc actcctcagg tgcaggctgc ctatcagaag
3241 gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct
3301 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
3361 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
3421 tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca
3481 tatgccatat gctggctgcc atgaacaaag gtggctataa agaggtcatc agtatatgaa
3541 acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt
3601 ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt
3661 tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct
3721 cttatgaaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc
3781 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
3841 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
3901 ccgctttcca gtcgggaaac ctgtcgtgcc agcggatcga tccgctgcat taatgaatcg
3961 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
4021 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
4081 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
4141 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
4201 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
4261 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
4321 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
4381 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
4441 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
4501 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
4561 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
4621 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
4681 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
4741 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
4801 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
4861 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
4921 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
4981 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
5041 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
5101 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
5161 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
5221 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
5281 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
5341 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
5401 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
5461 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
5521 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
5581 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
5641 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
5701 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
5761 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
5821 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
5881 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct g
//
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