| 质粒名称: | Flag Nprl2 pRK5 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 46333 |
| 存储实验室: | David Sabatini |
|
载体骨架基本信息 |
|
| 载体名称: | pRK5 |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | 5861 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | Nprl2型 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1143 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | 标志（主干上的N终端） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | SalI(未破坏) |
| 3’克隆位点: | NotI(未破坏) |
| 5’测序引物 | SP6 |
| 3’测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | Flag Nprl2 pRK5 was a gift from David Sabatini (Addgene plasmid # 46333 ; http://n2t.net/addgene:46333 ; RRID:Addgene_46333) |
| 参考文献部分: | A Tumor suppressor complex with GAP activity for the Rag GTPases that signal amino acid sufficiency to mTORC1. Bar-Peled L, Chantranupong L, Cherniack AD, Chen WW, Ottina KA, Grabiner BC, Spear ED, Carter SL, Meyerson M, Sabatini DM. Science. 2013 May 31;340(6136):1100-6. doi: 10.1126/science.1232044. 10.1126/science.1232044 PubMed 23723238 |
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 Flag Nprl2 pRK5 5861 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Flag Nprl2 pRK5
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5861)
AUTHORS Bar-Peled L, Chantranupong L, Cherniack AD, Chen WW, Ottina KA,
Grabiner BC, Spear ED, Carter SL, Meyerson M, Sabatini DM
TITLE A Tumor suppressor complex with GAP activity for the Rag GTPases
that signal amino acid sufficiency to mTORC1.
JOURNAL Science. 2013 May 31;340(6136):1100-6. doi: 10.1126/science.1232044.
PUBMED 23723238
REFERENCE 2 (bases 1 to 5861)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5861
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 581..605
/label=LNCX
/note="Human CMV promoter, forward primer"
promoter 815..833
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 815..832
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind 846..865
/label=pMT2-F
/note="Synthetic intron, forward primer"
CDS 924..947
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
regulatory 2148..2157
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
polyA_signal 2161..2295
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2198..2217)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2252..2271
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
promoter 2364..2721
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2387..2407)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2572..2707
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2634..2653
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(2741..2758)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(2741..2757)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2750..2772)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 2970..3425
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3057..3076)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3267..3288
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3442..3461)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3561..3583
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(3621..3639)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 3707..3811
/gene="bla"
/label=AmpR promoter
CDS 3812..4672
/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(4030..4049)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4843..5431
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5332..5351
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5585..5602
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 5719..5740
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5755..5785
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5793..5809
/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 5798..5820
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 5817..5833
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 5817..5833
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat
901 cgattgaatt caccatggaa atggactaca aggatgacga tgacaagggt ggtggtggtg
961 gtgcgtcgac gatgggcagc ggctgccgca tcgaatgcat attcttcagc gagttccacc
1021 ccacgctggg acccaagatc acctatcagg tccctgaaga cttcatctcc cgagagctgt
1081 ttgacacagt ccaagtgtac atcatcacca agccagagct gcagaacaag cttatcactg
1141 tcacagctat ggaaaagaag ctgatcggct gtcctgtgtg catcgaacac aagaagtaca
1201 gccgcaatgc tctcctcttc aacctgggct tcgtgtgtga tgcccaggcc aagacctgcg
1261 ccctcgagcc cattgttaaa aagctggctg gctatctgac cacactagag ctagagagca
1321 gcttcgtgtc catggaggag agcaagcaga agttggtgcc catcatgacc atcttgctgg
1381 aggagctaaa tgcctcaggc cggtgcactc tgcccattga tgagtccaac accatccact
1441 tgaaggtgat tgagcagcgg ccagaccctc cggtggccca ggagtatgat gtacctgtct
1501 ttaccaaaga caaggaggat ttcttcaact cacagtggga cctcactaca caacaaatcc
1561 tgccctacat tgatgggttc cgccacatcc agaagatttc agcagaggca gatgtggagc
1621 tcaacctggt gcgcattgct atccagaacc tgctgtacta cggcgttgtg acactggtgt
1681 ccatcctcca gtactccaat gtatactgcc caacgcccaa ggtccaggac ctggtagatg
1741 acaagtccct gcaagaggca tgtctatcct acgtgaccaa gcaagggcac aagagggcca
1801 gtctccggga tgtgttccag ctatactgca gcctgagccc tggcactacc gtgcgagacc
1861 tcattggccg ccacccccag cagctgcagc atgttgatga acggaagctg atccagttcg
1921 ggcttatgaa gaacctcatc aggcgactac agaagtatcc tgtgcgggtg actcgggaag
1981 agcagagcca ccctgcccgg ctttatacag gctgccacag ctatgacgag atctgctgca
2041 agacaggcat gagctaccat gagctggatg agcggcttga aaatgacccc aacatcatca
2101 tctgctggaa gtgagcggcc gctaagtaag taaggatccc cagcttggcc gccatggccc
2161 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
2221 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
2281 tatcatgtct ggatcgggaa ttaattcggc gcagcaccat ggcctgaaat aacctctgaa
2341 agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
2401 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
2461 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
2521 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
2581 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
2641 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
2701 ggaggcctag gcttttgcaa aaagctgtta acagcttggc actggccgtc gttttacaac
2761 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
2821 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
2881 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
2941 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
3001 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
3061 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
3121 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
3181 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
3241 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
3301 ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
3361 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
3421 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
3481 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
3541 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
3601 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
3661 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
3721 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
3781 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
3841 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
3901 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
3961 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
4021 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
4081 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
4141 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
4201 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
4261 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
4321 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
4381 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
4441 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
4501 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
4561 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
4621 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
4681 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
4741 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
4801 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
4861 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
4921 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
4981 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
5041 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
5101 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
5161 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
5221 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
5281 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
5341 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
5401 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
5461 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
5521 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
5581 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
5641 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
5701 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
5761 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
5821 aaacagctat gacatgatta cgaattaatt cgagctcgcc c
//
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