| 质粒名称: | pDest-636 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 159574 |
| 存储实验室: | Dominic Esposito |
|
载体骨架基本信息 |
|
| 载体名称: | -- |
| 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: | 昆虫表达 |
| 筛选标记: | -- |
| 载体抗性: | 氯霉素和庆大霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | ccdB Survival |
| 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: | -- |
| 标签/融合蛋白: | His6 MBP（插入件上的N端子） |
| 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: | pDest-636 was a gift from Dominic Esposito (Addgene plasmid # 159574 ; http://n2t.net/addgene:159574 ; RRID:Addgene_159574) |
| 参考文献部分: | Farnesylated and methylated KRAS4b: high yield production of protein suitable for biophysical studies of prenylated protein-lipid interactions. Gillette WK, Esposito D, Abreu Blanco M, Alexander P, Bindu L, Bittner C, Chertov O, Frank PH, Grose C, Jones JE, Meng Z, Perkins S, Van Q, Ghirlando R, Fivash M, Nissley DV, McCormick F, Holderfield M, Stephen AG. Sci Rep. 2015 Nov 2;5:15916. doi: 10.1038/srep15916. 10.1038/srep15916 PubMed 26522388 |
买家导航
pDest636载体基本信息
pDest636载体图谱质粒图谱和多克隆位点信息
pDest636载体简介
pDest636载体序列
LOCUS pDest-636 7689 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Gateway Destination vector for protein expression in baculovirus.
ACCESSION .
VERSION .
KEYWORDS pDest-636
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7689)
AUTHORS Gillette WK, Esposito D, Abreu Blanco M, Alexander P, Bindu L,
Bittner C, Chertov O, Frank PH, Grose C, Jones JE, Meng Z, Perkins
S, Van Q, Ghirlando R, Fivash M, Nissley DV, McCormick F,
Holderfield M, Stephen AG
TITLE Farnesylated and methylated KRAS4b: high yield production of protein
suitable for biophysical studies of prenylated protein-lipid
interactions.
JOURNAL Sci Rep. 2015 Nov 2;5:15916. doi: 10.1038/srep15916.
PUBMED 26522388
REFERENCE 2 (bases 1 to 7689)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7689
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..92
/gene="polh from Autographa californica"
/label=polyhedrin promoter
/note="promoter for the baculovirus polyhedrin gene"
primer_bind 17..34
/label=Polyhedrin-F
/note="Polyhedrin promoter, forward primer"
CDS 146..163
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 167..1264
/codon_start=1
/gene="malE (mutated)"
/product="maltose binding protein from E. coli"
/label=MBP
/note="This version of the gene does not encode a signal
sequence, so MBP will remain in the cytosol."
/translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
YAVRTAVINAASGRQTVDEALKDAQT"
regulatory 1121..1130
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
primer_bind 1238..1261
/label=MBP-F
/note="Maltose binding protein, forward primer"
protein_bind 1283..1407
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 1432..1462
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 1516..2175
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(1582..1601)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
CDS 2517..2822
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
primer_bind 2701..2720
/label=ccdB-fwd
/note="ccdB gene, forward primer"
protein_bind complement(2863..2987)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
polyA_signal 3132..3266
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3169..3188)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3223..3242
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
mobile_element 3295..3460
/mobile_element_type="transposon:Tn7"
/note="mini-Tn7 element (left end of the Tn7 transposon)"
rep_origin 3644..4098
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3731..3750)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3940..3961
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 4124..4228
/gene="bla"
/label=AmpR promoter
CDS 4229..5089
/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(4447..4466)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5260..5848
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5749..5768
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6002..6019
/label=L4440
/note="L4440 vector, forward primer"
mobile_element 6153..6377
/mobile_element_type="transposon:Tn7"
/note="mini-Tn7 element (right end of the Tn7 transposon)"
CDS complement(6444..6977)
/codon_start=1
/gene="aacC1"
/product="gentamycin acetyltransferase"
/label=GmR
/note="confers resistance to gentamycin"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(7166..7194)
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
promoter complement(7557..7666)
/label=p10 promoter
/note="baculovirus promoter for expression in insect cells"
ORIGIN
1 atcatggaga taattaaaat gataaccatc tcgcaaataa ataagtattt tactgttttc
61 gtaacagttt tgtaataaaa aaacctataa atattccgga ttattcatac cgtcccacca
121 tcgggcgcgg atcaccatgg gatcccatca ccatcaccat cacggcaaaa tcgaagaagg
181 taaactggta atctggatta acggcgataa aggctataac ggtctcgctg aagtcggtaa
241 gaaattcgag aaagataccg gaattaaagt caccgttgag catccggata aactggaaga
301 gaaattccca caggttgcgg caactggcga tggccctgac attatcttct gggcacacga
361 ccgctttggt ggctacgctc aatctggcct gttggctgaa atcaccccgg acaaagcgtt
421 ccaggacaag ctgtatccgt ttacctggga tgccgtacgt tacaacggca agctgattgc
481 ttacccgatc gctgttgaag cgttatcgct gatttataac aaagatctgc tgccgaaccc
541 gccaaaaacc tgggaagaga tcccggcgct ggataaagaa ctgaaagcga aaggtaagag
601 cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg ccgctgattg ctgctgacgg
661 gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa
721 cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg attaaaaaca aacacatgaa
781 tgcagacacc gattactcca tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac
841 catcaacggc ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt atggtgtaac
901 ggtactgccg accttcaagg gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg
961 tattaacgct gccagtccga acaaagagct ggcaaaagag ttcctcgaaa actatctgct
1021 gactgatgaa ggtctggaag cggttaataa agacaaaccg ctgggtgccg tagcgctgaa
1081 gtcttacgag gaagagttgg cgaaagatcc acgtattgcc gccaccatgg aaaacgccca
1141 gaaaggtgaa atcatgccga acatcccgca gatgtccgct ttctggtatg ccgtgcgtac
1201 tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat gaagccctga aagacgcgca
1261 gactaattcg ggatcctcga tcacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa
1321 tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta cataatactg
1381 taaaacacaa catatccagt cactatggcg gccgcattag gcaccccagg ctttacactt
1441 tatgcttccg gctcgtataa tgtgtggatt ttgagttagg atccgtcgag attttcagga
1501 gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa
1561 tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag
1621 accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt
1681 tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg
1741 gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc
1801 catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag
1861 tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct
1921 aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt
1981 tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa
2041 tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt
2101 tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg
2161 cagggcgggg cgtaaacgcg tggatccggc ttactaaaag ccagataaca gtatgcgtat
2221 ttgcgcgctg atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca
2281 aaaagaggta tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt
2341 tgctcaaggc atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga
2401 agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt
2461 cgcccggttt attgaaatga acggctcttt tgctgacgag aacaggggct ggtgaaatgc
2521 agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga
2581 gtgatattat tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc
2641 tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc
2701 gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg
2761 atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat
2821 aaatgtcagg ctcccttata cacagccagt ctgcaggtcg accatagtga ctggatatgt
2881 tgtgttttac agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata
2941 tttatatcat tttacgtttc tcgttcagct ttcttgtaca aagtggtgat cgagctagct
3001 agaagcttgt cgagaagtac tagaggatca taatcagcca taccacattt gtagaggttt
3061 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
3121 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
3181 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
3241 tcaatgtatc ttatcatgtc tggatctgat cactgcttga gcctaggaga tccgaaccag
3301 ataagtgaaa tctagttcca aactattttg tcatttttaa ttttcgtatt agcttacgac
3361 gctacaccca gttcccatct attttgtcac tcttccctaa ataatcctta aaaactccat
3421 ttccacccct cccagttccc aactattttg tccgcccaca gcggggcatt tttcttcctg
3481 ttatgttttt aatcaaacat cctgccaact ccatgtgaca aaccgtcatc ttcggctact
3541 ttttctctgt cacagaatga aaatttttct gtcatctctt cgttattaat gtttgtaatt
3601 gactgaatat caacgcttat ttgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
3661 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
3721 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
3781 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
3841 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
3901 gtttttcgcc ctttgacgtt ggagtccacg ttcttaatag tggactcttg ttccaaactg
3961 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
4021 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
4081 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
4141 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
4201 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
4261 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
4321 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
4381 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
4441 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
4501 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
4561 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
4621 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
4681 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
4741 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
4801 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
4861 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
4921 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
4981 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
5041 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
5101 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
5161 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
5221 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
5281 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
5341 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
5401 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
5461 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
5521 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
5581 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
5641 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
5701 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
5761 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
5821 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
5881 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
5941 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
6001 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
6061 tctgtgcggt atttcacacc gcatagacca gccgcgtaac ctggcaaaat cggttacggt
6121 tgagtaataa atggatgccc tgcgtaagcg ggtgtgggcg gacaataaag tcttaaactg
6181 aacaaaatag atctaaacta tgacaataaa gtcttaaact agacagaata gttgtaaact
6241 gaaatcagtc cagttatgct gtgaaaaagc atactggact tttgttatgg ctaaagcaaa
6301 ctcttcattt tctgaagtgc aaattgcccg tcgtattaaa gaggggcgtg gccaagggca
6361 tggtaaagac tatattcgcg gcgttgtgac aatttaccga acaactccgc ggccgggaag
6421 ccgatctcgg cttgaacgaa ttgttaggtg gcggtacttg ggtcgatatc aaagtgcatc
6481 acttcttccc gtatgcccaa ctttgtatag agagccactg cgggatcgtc accgtaatct
6541 gcttgcacgt agatcacata agcaccaagc gcgttggcct catgcttgag gagattgatg
6601 agcgcggtgg caatgccctg cctccggtgc tcgccggaga ctgcgagatc atagatatag
6661 atctcactac gcggctgctc aaacttgggc agaacgtaag ccgcgagagc gccaacaacc
6721 gcttcttggt cgaaggcagc aagcgcgatg aatgtcttac tacggagcaa gttcccgagg
6781 taatcggagt ccggctgatg ttgggagtag gtggctacgt ctccgaactc acgaccgaaa
6841 agatcaagag cagcccgcat ggatttgact tggtcagggc cgagcctaca tgtgcgaatg
6901 atgcccatac ttgagccacc taactttgtt ttagggcgac tgccctgctg cgtaacatcg
6961 ttgctgctgc gtaacatcgt tgctgctcca taacatcaaa catcgaccca cggcgtaacg
7021 cgcttgctgc ttggatgccc gaggcataga ctgtacaaaa aaacagtcat aacaagccat
7081 gaaaaccgcc actgcgccgt taccaccgct gcgttcggtc aaggttctgg accagttgcg
7141 tgagcgcata cgctacttgc attacagttt acgaaccgaa caggcttatg tcaactgggt
7201 tcgtgccttc atccgtttcc acggtgtgcg tcacccggca accttgggca gcagcgaagt
7261 cgaggcattt ctgtcctggc tggcgaacga gcgcaaggtt tcggtctcca cgcatcgtca
7321 ggcattggcg gccttgctgt tcttctacgg caaggtgctg tgcacggatc tgcccttgct
7381 tcaggagatc ggtagacctc ggccgtcgcg gcgcttgccg gtggtgctga ccccggatga
7441 agtggttcgc atcctcggtt ttctggaagg cgagcatcgt ttgttcgccc aggactctag
7501 ctatagttct agtggttggc tactcgagat cccgggtgat caagtcttcg tcgagtgatt
7561 gtaaataaaa tgtaatttac agtatagtat tttaattaat atacaaatga tttgataata
7621 attcttattt aactataata tattgtgttg ggttgaatta aaggtccgta tactccggaa
7681 tattaatag
//
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