| 质粒名称: | pRK5-FLAG-C7orf59 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 42332 |
| 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: | BD PharMingen |
| 空载体大小: | 4661 bp |
| 完整质粒大小: | 5018 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: | C7orf59 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | NM_001008395.2 NP_001008396.1 |
| 启动子: | CMV, SP6 |
| 标签/融合蛋白: | 标志（插入时的N端子） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoRI (未破坏) |
| 3’克隆位点: | HindIII (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: | pRK5-FLAG-C7orf59 was a gift from David Sabatini (Addgene plasmid # 42332 ; http://n2t.net/addgene:42332 ; RRID:Addgene_42332) |
| 参考文献部分: | Ragulator Is a GEF for the Rag GTPases that Signal Amino Acid Levels to mTORC1. Bar-Peled L, Schweitzer LD, Zoncu R, Sabatini DM. Cell. 2012 Sep 14;150(6):1196-208. doi: 10.1016/j.cell.2012.07.032. 10.1016/j.cell.2012.07.032 PubMed 22980980 |
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-FLAG-C7orf59 5018 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pRK5-FLAG-C7orf59
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5018)
AUTHORS Bar-Peled L, Schweitzer LD, Zoncu R, Sabatini DM
TITLE Ragulator Is a GEF for the Rag GTPases that Signal Amino Acid Levels
to mTORC1.
JOURNAL Cell. 2012 Sep 14;150(6):1196-208. doi: 10.1016/j.cell.2012.07.032.
PUBMED 22980980
REFERENCE 2 (bases 1 to 5018)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5018
/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 1305..1314
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
polyA_signal 1318..1452
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1355..1374)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1409..1428
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
promoter 1521..1878
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(1544..1564)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 1729..1864
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 1791..1810
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(1898..1915)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(1898..1914)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1907..1929)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 2127..2582
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2214..2233)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2424..2445
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2599..2618)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2718..2740
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(2778..2796)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 2864..2968
/gene="bla"
/label=AmpR promoter
CDS 2969..3829
/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(3187..3206)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4000..4588
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4489..4508
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4742..4759
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 4876..4897
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4912..4942
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4950..4966
/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 4955..4977
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 4974..4990
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 4974..4990
/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 gatgacttct gcgctgaccc aggggctgga gcgaatccca gaccagctcg
1021 gctacctggt actgagtgaa ggtgcagtgc tggcgtcatc tggggacctg gagaatgatg
1081 agcaggcagc cagtgccatc tctgagctgg tcagcacagc ctgcggtttc cggctgcacc
1141 gcggcatgaa tgtgcccttc aagcgcctgt ctgtggtctt tggagaacac acactgctgg
1201 tgacggtgtc aggacagagg gtgtttgtgg tgaagaggca gaaccgaggt cgggagccca
1261 ttgatgtctg agcggccgct aagtaagtaa ggatccccag cttggccgcc atggcccaac
1321 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
1381 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
1441 catgtctgga tcgggaatta attcggcgca gcaccatggc ctgaaataac ctctgaaaga
1501 ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
1561 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
1621 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
1681 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
1741 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
1801 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
1861 ggcctaggct tttgcaaaaa gctgttaaca gcttggcact ggccgtcgtt ttacaacgtc
1921 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
1981 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
2041 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
2101 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
2161 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
2221 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
2281 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
2341 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
2401 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
2461 tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
2521 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
2581 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
2641 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
2701 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
2761 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
2821 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
2881 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
2941 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
3001 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
3061 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
3121 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
3181 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
3241 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
3301 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
3361 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
3421 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
3481 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
3541 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
3601 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
3661 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
3721 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
3781 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
3841 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
3901 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
3961 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
4021 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
4081 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
4141 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
4201 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
4261 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
4321 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
4381 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
4441 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
4501 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
4561 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
4621 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
4681 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
4741 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
4801 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
4861 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
4921 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
4981 cagctatgac atgattacga attaattcga gctcgccc
//
