| 质粒名称: | pRK5-HA-Ubiquitin-K6 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 22900 |
| 存储实验室: | Sandra Weller |
|
载体骨架基本信息 |
|
| 载体名称: | pRK5-HA |
| 空载体大小: | 4800 bp |
| 完整质粒大小: | 4996 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 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: | Ubiquitin C / Ub |
| 物种来源: | H. sapiens (human) |
| 插入大小： | 220bp |
| 基因突变: | K6 only, all other lysines mutated to arginines |
| 标签/融合蛋白: | HA (N terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | unknown (unknown if destroyed) |
| 3’克隆位点: | unknown (unknown if destroyed) |
| 5’测序引物 | SP6 |
|
文献引用方法 |
|
| 材料和方法部分: |
pRK5-HA-Ubiquitin-K6 was a gift from Sandra Weller (Addgene plasmid # 22900 ; http://n2t.net/addgene:22900 ; RRID:Addgene_22900)
|
| 参考文献部分: | Virus-Induced Chaperone-Enriched (VICE) domains function as nuclear protein quality control centers during HSV-1 infection. Livingston CM, Ifrim MF, Cowan AE, Weller SK. PLoS Pathog. 2009 Oct . 5(10):e1000619. 10.1371/journal.ppat.1000619 PubMed 19816571 |
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 pRK5-HA-Ubiquitin-K6 4996 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pRK5-HA-Ubiquitin-K6
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4996)
AUTHORS Livingston CM, Ifrim MF, Cowan AE, Weller SK
TITLE Virus-Induced Chaperone-Enriched (VICE) domains function as nuclear
protein quality control centers during HSV-1 infection.
JOURNAL PLoS Pathog. 2009 Oct . 5(10):e1000619.
PUBMED 19816571
REFERENCE 2 (bases 1 to 4996)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4996
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 98..119
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 134..164
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 172..188
/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)."
primer_bind 177..199
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 196..212
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 196..212
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
enhancer 241..620
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 621..824
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 775..795
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 821..845
/label=LNCX
/note="Human CMV promoter, forward primer"
promoter 1055..1073
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 1055..1072
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind 1086..1105
/label=pMT2-F
/note="Synthetic intron, forward primer"
regulatory 1190..1199
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1202..1228
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 1259..1486
/codon_start=1
/product="human ubiquitin variant containing Lys-6, with
all other lysines mutated to arginine"
/label=ubiquitin-K6
/translation="MQIFVKTLTGRTITLEVEPSDTIENVRARIQDREGIPPDQQRLIF
AGRQLEDGRTLSDYNIQRESTLHLVLRLRGG"
regulatory 1523..1532
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
polyA_signal 1536..1670
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1573..1592)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1627..1646
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
promoter 1739..2096
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(1762..1782)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 1947..2082
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2009..2028
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(2116..2133)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(2116..2132)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2125..2147)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 2345..2800
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2432..2451)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2642..2663
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2817..2836)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2936..2958
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(2996..3014)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 3082..3186
/gene="bla"
/label=AmpR promoter
CDS 3187..4047
/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(3405..3424)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4218..4806
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4707..4726
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4960..4977
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
61 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
121 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
181 gataacaatt tcacacagga aacagctatg acatgattac gaattaattc gagctcgccc
241 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
301 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
361 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
421 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
481 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
541 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
601 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
661 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
721 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
781 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
841 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
901 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
961 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
1021 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
1081 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat
1141 cgattgaatt cccgaaccga cagtcggtct cttcaccaag gccattcgcg ccaccatggg
1201 ctacccctat gatgtgcctg actacgcaga tctcaatggt ggtggtggtg ggtcgaccat
1261 gcagatcttc gtcaaaacgt taaccggtag aaccataact ctagaagttg aaccatccga
1321 taccatcgaa aacgttagag ctagaattca agacagagaa ggcattccac ctgatcaaca
1381 aagattgatc tttgccggta gacagctcga ggacggtaga acgctgtctg attacaacat
1441 tcagagagag tcgaccttac atcttgtctt aagactaaga ggtggttgag cggccgctaa
1501 gtaagtaagg atccccagct tggccgccat ggcccaactt gtttattgca gcttataatg
1561 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
1621 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc gggaattaat
1681 tcggcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta ggtaccttct
1741 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
1801 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
1861 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
1921 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
1981 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
2041 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
2101 tgttaacagc ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
2161 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
2221 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat
2281 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
2341 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
2401 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
2461 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
2521 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
2581 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
2641 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttaat
2701 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
2761 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
2821 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
2881 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
2941 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
3001 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
3061 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
3121 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
3181 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
3241 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
3301 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
3361 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
3421 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
3481 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
3541 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
3601 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
3661 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
3721 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
3781 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
3841 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
3901 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
3961 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
4021 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
4081 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
4141 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
4201 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
4261 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
4321 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
4381 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
4441 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
4501 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
4561 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
4621 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
4681 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
4741 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
4801 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
4861 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
4921 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
4981 gcggaagagc gcccaa
//
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