| 质粒名称: | HA Nprl3 pRK5 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 46330 |
| 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: | 6475 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: | Nprl3 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1710 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | HA（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: | BamHI(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: | HA Nprl3 pRK5 was a gift from David Sabatini (Addgene plasmid # 46330 ; http://n2t.net/addgene:46330 ; RRID:Addgene_46330) |
| 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 HA Nprl3 pRK5 6475 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS HA Nprl3 pRK5
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6475)
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 6475)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6475
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 107..128
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/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 186..208
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 205..221
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
enhancer 250..629
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 630..833
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 784..804
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 830..854
/label=LNCX
/note="Human CMV promoter, forward primer"
promoter 1064..1082
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 1064..1081
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind 1095..1114
/label=pMT2-F
/note="Synthetic intron, forward primer"
regulatory 1199..1208
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1211..1237
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
regulatory 3011..3020
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
polyA_signal 3024..3158
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3061..3080)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3115..3134
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
promoter 3227..3584
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3250..3270)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3435..3570
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3497..3516
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(3604..3621)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(3604..3620)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3613..3635)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 3833..4288
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3920..3939)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4130..4151
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4305..4324)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4424..4446
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4484..4502)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4570..4674
/gene="bla"
/label=AmpR promoter
CDS 4675..5535
/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(4893..4912)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5706..6294
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6195..6214
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6448..6465
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
181 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aattaattcg
241 agctcgcccg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
301 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
361 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
421 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
481 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
541 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
601 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
661 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
721 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
781 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
841 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
901 gggaccgatc cagcctccgc ggccgggaac ggtgcattgg aacgcggatt ccccgtgcca
961 agagtgacgt aagtaccgcc tatagagtct ataggcccac ccccttggct tcgttagaac
1021 gcggctacaa ttaatacata accttatgta tcatacacat acgatttagg tgacactata
1081 gaataacatc cactttgcct ttctctccac aggtgtccac tcccaggtcc aactgcacct
1141 cggttctatc gattgaattc ccgaaccgac agtcggtctc ttcaccaagg ccattcgcgc
1201 caccatgggc tacccctatg atgtgcctga ctacgcagat ctcaatggtg gtggtggtgg
1261 gtcgacgatg cgggacaaca ccagccccat cagcgtgatt ctggtgagct cggggagcag
1321 gggcaataag ctgctgttca ggtacccctt ccagagaagc caggagcacc cggcgtccca
1381 gacaagtaag ccgcgtagca gatacgctgc cagcaacacg ggcgaccatg ctgatgagca
1441 ggacggcgat tccaggtttt cagatgttat tctggcaaca attttggcaa ccaagtctga
1501 aatgtgtggc caaaaatttg aactgaagat tgataatgtg cgatttgttg ggcacccaac
1561 actgctacag catgctctgg ggcagatctc caaaacagat ccttccccga agagggaagc
1621 acctactatg attcttttta atgtggtgtt tgcactgagg gccaacgcag acccgtcagt
1681 gataaactgt ctgcataacc tgtcccgtcg tatcgccacc gtgctgcagc acgaggagcg
1741 ccgctgccag tacctcaccc gggaggccaa gctgatcctg gcgctccagg atgaggtgtc
1801 cgccatggct gatggaaatg aaggtcctca gtccccattc catcacatcc tgcccaagtg
1861 caagctggcc agggacctca aggaagctta tgacagcctg tgcacgtcgg gcgtagttcg
1921 gcttcacatc aacagctggc tggaggtgag cttctgcctg ccccacaaga tccactatgc
1981 ggcctccagt ctgatccccc cagaggccat cgaacggagc ctgaaagcca tccgccccta
2041 ccatgccctg ctgctgctca gtgatgagaa gtccttgctg ggtgagcttc ctattgactg
2101 ctcccctgcc ctagtgcggg tgatcaagac cacatctgct gtgaagaacc tgcagcagct
2161 agcccaagat gcggacctgg ccttgctgca ggttttccag cttgcagctc atctggtgta
2221 ctggggcaag gccatcatca tctacccgct gtgtgagaac aacgtctaca tgctgtctcc
2281 caatgccagc gtatgtctgt actccccgct ggccgagcag ttctcccacc agttcccatc
2341 tcatgacctg ccgtccgttc ttgccaagtt ctccttgccg gtctccttgt cagaatttag
2401 gaatcccctg gcccccgctg tgcaggagac ccagctcatc cagatggtgg tgtggatgct
2461 gcagcgccgg cttctcatcc agctgcacac ctatgtctgc ctgatggcct cacccagcga
2521 ggaggagccc cgtccgcgag aggacgacgt ccccttcact gcccgggtcg gcggtcgcag
2581 cctcagcacg cccaacgccc tcagctttgg ctccccaacc agcagcgatg acatgaccct
2641 caccagcccc agcatggaca actccagcgc agagctactt cccagcgggg actcgccact
2701 gaaccagagg atgacggaga acctgctggc cagcctgtcg gagcatgaac gcgcagccat
2761 cctcagtgta cccgcagccc agaaccctga ggacctccgc atgtttgcca ggctccttca
2821 ctacttccgc ggacgccacc acctggagga gattatgtac aacgagaaca cgcggcgctc
2881 ccagctgctc atgctgtttg acaagttccg cagcgtgctg gtggtgacca cccacgagga
2941 ccctgtcatt gccgtcttcc aggctctgct cccctgagcg gccgctaagt aagtaaggat
3001 ccccagcttg gccgccatgg cccaacttgt ttattgcagc ttataatggt tacaaataaa
3061 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
3121 tgtccaaact catcaatgta tcttatcatg tctggatcgg gaattaattc ggcgcagcac
3181 catggcctga aataacctct gaaagaggaa cttggttagg taccttctga ggcggaaaga
3241 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
3301 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
3361 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
3421 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
3481 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
3541 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctg ttaacagctt
3601 ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
3661 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
3721 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct
3781 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
3841 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
3901 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
3961 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
4021 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
4081 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
4141 ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt
4201 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
4261 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
4321 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
4381 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
4441 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
4501 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
4561 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
4621 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
4681 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
4741 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
4801 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
4861 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
4921 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
4981 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
5041 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
5101 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
5161 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
5221 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
5281 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
5341 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
5401 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
5461 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
5521 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
5581 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
5641 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
5701 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
5761 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
5821 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
5881 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
5941 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
6001 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
6061 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
6121 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
6181 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
6241 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
6301 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
6361 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
6421 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaag
//
