| 质粒名称: | pcDNA3/Flag-METTL14 |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 53740 |
| 存储实验室: | Chuan He |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3 |
| 载体出品公司: | Invitrogen |
| 空载体大小: | 5446 bp |
| 完整质粒大小: | 6710bp |
| 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: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | Methyltransferase-like 14 / METTL14 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1371bp |
| GenBank编号 | NM_020961.2 |
| 启动子: | T7 |
| 标签/融合蛋白: | FLAG tag (N terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BamHI(未破坏) |
| 3’克隆位点: | XhoI (未破坏) |
| 5’测序引物 | 5' TAATACGACTCACTATAGGG 3' |
| 3’测序引物 | 5' ATTTAGGTGACACTATAG 3' |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3/Flag-METTL14 was a gift from Chuan He (Addgene plasmid # 53740 ; http://n2t.net/addgene:53740 ; RRID:Addgene_53740) |
| 参考文献部分: | A METTL3-METTL14 complex mediates mammalian nuclear RNA N6-adenosine methylation. Liu J, Yue Y, Han D, Wang X, Fu Y, Zhang L, Jia G, Yu M, Lu Z, Deng X, Dai Q, Chen W, He C. Nat Chem Biol. 2014 Feb;10(2):93-5. doi: 10.1038/nchembio.1432. Epub 2013 Dec 6. 10.1038/nchembio.1432 PubMed 24316715 |
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 pcDNA3/Flag-METTL14 6710 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION For expression of METTL14 (methyltransferase-like 14) in human
cells.
ACCESSION .
VERSION .
KEYWORDS pcDNA3/Flag-METTL14
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6710)
AUTHORS Liu J, Yue Y, Han D, Wang X, Fu Y, Zhang L, Jia G, Yu M, Lu Z, Deng
X, Dai Q, Chen W, He C
TITLE A METTL3-METTL14 complex mediates mammalian nuclear RNA N6-adenosine
methylation.
JOURNAL Nat Chem Biol. 2014 Feb;10(2):93-5. doi: 10.1038/nchembio.1432. Epub
2013 Dec 6.
PUBMED 24316715
REFERENCE 2 (bases 1 to 6710)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6710
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 74..453
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 454..657
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 608..628
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 702..721
/label=T7
/note="T7 promoter, forward primer"
promoter 702..720
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 755..778
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
promoter complement(2174..2192)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(2175..2192)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(2212..2229)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2218..2442
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2488..2916
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2575..2594)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2785..2806
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2925..2945)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2969..3179
/label=SV40 promoter
/note="SV40 early promoter"
promoter 2991..3187
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 3038..3173
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3100..3119
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3254..4048
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3308..3327)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3918..3937
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4222..4343
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4259..4278)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4313..4332
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4392..4408)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4392..4408)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4405..4427)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4416..4432
/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)."
promoter complement(4440..4470)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4485..4506
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4623..4640)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4794..5382)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4874..4893)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5553..6413)
/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 6176..6195
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6414..6518)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6593..6612)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
61 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
121 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
181 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
241 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
301 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
361 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
421 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
481 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
541 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
601 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc
661 tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg
721 gagacccaag cttggtaccg agctcggatc catggattac aaggatgacg atgacaagga
781 tagccgcttg caggagatcc gggagcggca gaagttacgg cgacagctcc tcgcgcagca
841 gttgggagct gaaagtgccg acagcattgg tgccgtgtta aatagcaaag atgagcagag
901 agaaattgct gaaacaagag aaacttgcag ggcttcctat gatacctctg ctccaaatgc
961 aaaacgtaag tatctggatg aaggagagac agatgaagac aaaatggaag aatataagga
1021 tgaactagaa atgcaacagg atgaagaaaa tttgccatat gaagaagaga tttacaaaga
1081 ttctagtact tttcttaagg gaacacagag cttaaatccc cataatgatt actgccaaca
1141 ttttgtagac actggacata gacctcagaa tttcatcagg gatgtaggtt tagctgacag
1201 atttgaagaa tatcctaaac tgagggagct catcaggcta aaggatgagt taatagctaa
1261 atctaacact cctcccatgt acttacaagc cgatatagaa gcctttgaca tcagagaact
1321 aacacccaaa tttgatgtga ttcttctgga acccccttta gaagaatatt acagagaaac
1381 tggcatcact gctaatgaaa aatgctggac ttgggatgat attatgaagt tagaaattga
1441 tgagattgca gcacctcgat catttatttt tctctggtgt ggttctgggg aggggttgga
1501 ccttggaaga gtgtgtttac gaaaatgggg ttacagaaga tgtgaagata tttgttggat
1561 taaaaccaat aaaaacaatc ctgggaagac taagacttta gatccaaagg ctgtctttca
1621 gagaacaaag gaacactgcc tcatggggat caaaggaact gtgaagcgta gcacagacgg
1681 ggacttcatt catgctaatg ttgacattga cttaattatc acagaagaac ctgaaattgg
1741 caatatagaa aaacctgtag aaatttttca tataattgag catttttgtc ttggtagaag
1801 acgccttcat ctatttggaa gagatagtac aattcgacca ggctggctca cagttggacc
1861 aacgcttaca aatagcaact acaatgcaga aacatatgca tcctatttca gtgctcctaa
1921 ttcctacttg actggttgta cagaagaaat tgagagactt cgaccaaaat cgcctcctcc
1981 caaatctaaa tctgaccgag gaggtggagc tcccagaggt ggaggaagag gtggaacttc
2041 tgctggccgt ggacgagaaa gaaatagatc taacttccga ggagaaagag gtggctttag
2101 agggggccgt ggaggagcac acagaggtgg ctttccacct cgataactcg agcatgcatc
2161 tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg
2221 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
2281 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
2341 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
2401 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
2461 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
2521 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
2581 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
2641 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
2701 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
2761 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
2821 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
2881 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
2941 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
3001 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
3061 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
3121 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
3181 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
3241 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
3301 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
3361 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
3421 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
3481 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
3541 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
3601 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
3661 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
3721 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
3781 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
3841 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
3901 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
3961 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
4021 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
4081 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
4141 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
4201 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
4261 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
4321 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
4381 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
4441 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
4501 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
4561 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
4621 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
4681 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
4741 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
4801 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
4861 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
4921 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
4981 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
5041 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
5101 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
5161 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
5221 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
5281 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
5341 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
5401 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
5461 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
5521 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
5581 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
5641 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
5701 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
5761 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
5821 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
5881 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
5941 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
6001 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
6061 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
6121 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
6181 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
6241 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
6301 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
6361 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
6421 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
6481 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
6541 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
6601 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
6661 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
//
