| 质粒名称: | pnEA-NpHG_N |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 147033 |
| 存储实验室: | Elisa Izaurralde |
|
载体骨架基本信息 |
|
| 载体名称: | pnEA |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 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: | 低拷贝 |
|
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: | -- |
| 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: | pnEA-NpHG_N was a gift from Elisa Izaurralde (Addgene plasmid # 147033 ; http://n2t.net/addgene:147033 ; RRID:Addgene_147033) |
| 参考文献部分: | Deciphering correct strategies for multiprotein complex assembly by co-expression: application to complexes as large as the histone octamer. Diebold ML, Fribourg S, Koch M, Metzger T, Romier C. J Struct Biol. 2011 Aug;175(2):178-88. doi: 10.1016/j.jsb.2011.02.001. Epub 2011 Feb 12. 10.1016/j.jsb.2011.02.001 PubMed 21320604 |
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 pnEA-NpHG_N 6434 bp ds-DNA circular SYN 25-FEB-2022
DEFINITION Bacterial Expression vector.
ACCESSION .
VERSION .
KEYWORDS pnEA-NpHG_N
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6434)
AUTHORS Diebold ML, Fribourg S, Koch M, Metzger T, Romier C
TITLE Deciphering correct strategies for multiprotein complex assembly by
co-expression: application to complexes as large as the histone
octamer.
JOURNAL J Struct Biol. 2011 Aug;175(2):178-88. doi:
10.1016/j.jsb.2011.02.001. Epub 2011 Feb 12.
PUBMED 21320604
REFERENCE 2 (bases 1 to 6434)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Feb 25, 2022 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6434
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(18..36)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 104..208
/gene="bla"
/label=AmpR promoter
CDS 209..1069
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(427..446)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1240..1828
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1729..1748
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1982..1999
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 2014..2153
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(2079..2098)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2239..2261
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(2255..2446)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
primer_bind 4801..4820
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(4840..4917)
/gene="lacI"
/label=lacI promoter
primer_bind 5123..5142
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 5226..5245
/label=T7
/note="T7 promoter, forward primer"
promoter 5226..5244
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 5245..5269
/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)."
RBS 5284..5306
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 5325..5342
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 5358..6011
/codon_start=1
/product="glutathione S-transferase from Schistosoma
japonicum"
/label=GST
/translation="MAPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNSDHVTHPDFMLYDALDVVL
YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
primer_bind 5969..5991
/label=pGEX 5'
/note="pGEX vectors, Glutathione-S-transferase, forward
primer"
CDS 6033..6056
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
primer_bind complement(6161..6179)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 6175..6222
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter complement(6397..6425)
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
ORIGIN
1 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
61 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
181 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
421 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
721 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
1021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
1081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
1141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
1201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
1261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
1321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
1381 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
1441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
1501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
1561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
1621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
1681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
1741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
1801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
1861 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
1921 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
1981 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
2041 tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca
2101 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
2161 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
2221 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
2281 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
2341 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
2401 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
2461 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
2521 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg
2581 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
2641 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
2701 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
2761 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
2821 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
2881 tcaacgacag gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgagatgc
2941 gccgcgtgcg gctgctggag atggcggacg cgatggatat gttctgccaa gggttggttt
3001 gcgcattcac agttctccgc aagaattgat tggctccaat tcttggagtg gtgaatccgt
3061 tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc cggctccatg caccgcgacg
3121 caacgcgggg aggcagacaa ggtatagggc ggcgcctaca atccatgcca acccgttcca
3181 tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc ggtccaatga tcgaagttag
3241 gctggtaaga gccgcgagcg atccttgaag ctgtccctga tggtcgtcat ctacctgcct
3301 ggacagcatg gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa gaatcataat
3361 ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag acgtagccca gcgcgtcggc
3421 cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac
3481 gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt
3541 cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc
3601 tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc
3661 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc
3721 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg
3781 aaactgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
3841 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
3901 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
3961 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
4021 gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
4081 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgcccttatt
4141 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
4201 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
4261 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
4321 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
4381 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
4441 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
4501 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
4561 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
4621 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
4681 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
4741 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
4801 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
4861 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
4921 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
4981 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
5041 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
5101 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
5161 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc
5221 gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc cactagtaat
5281 aattttgttt aactttaaga aggagatata ccatgagcgg cagccatcat catcatcatc
5341 acagcggcag cggctccatg gcccctatac taggttattg gaaaattaag ggccttgtgc
5401 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
5461 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
5521 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
5581 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
5641 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
5701 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
5761 atcgtttatg tcataaaaca tatttaaata gtgatcatgt aacccatcct gacttcatgt
5821 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
5881 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
5941 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
6001 atccgccaaa atcggatggt accggcagcg gcctggaagt tctgttccag gggccccata
6061 tgctcgagct taagcaattg ggatcctaat agtctagagc tagccctagg agatccggct
6121 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
6181 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
6241 tccggatatc ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat
6301 ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct
6361 gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct
6421 gtcaaacatg agaa
//
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