| 质粒名称: | pFRT/TO/HIS/FLAG/HA-ALKBH5 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 38073 |
| 存储实验室: | Markus Landthaler |
|
载体骨架基本信息 |
|
| 载体名称: | pFRT/TO/HIS/FLAG/HA-DEST |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5522 bp |
| 完整质粒大小: | 6875 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 潮霉素 |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | ALKBH5 alkB, alkylation repair homolog 5 (E. coli),ABH5; OFOXD; OFOXD1 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1353 bp |
| 基因突变: | cDNA克隆MGC:71143图像：6339184，3'UTR较长，N端被截断：位置34-aaavaaa序列缺失 |
| GenBank编号 | NM_017758.3 |
| 启动子 | -- |
| 标签/融合蛋白: | HIS/FLAG/HA（主干上的N终端） |
| 克隆方法: | Gateway克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | CMV |
| 3' 测序引物 | BGH |
|
文献引用方法 |
|
| 材料和方法部分: | pFRT/TO/HIS/FLAG/HA-ALKBH5 was a gift from Markus Landthaler (Addgene plasmid # 38073 ; http://n2t.net/addgene:38073 ; RRID:Addgene_38073) |
| 参考文献部分: | The mRNA-Bound Proteome and Its Global Occupancy Profile on Protein-Coding Transcripts. Baltz AG, Munschauer M, Schwanhausser B, Vasile A, Murakawa Y, Schueler M, Youngs N, Penfold-Brown D, Drew K, Milek M, Wyler E, Bonneau R, Selbach M, Dieterich C, Landthaler M. Mol Cell. 2012 Jun 8;46(5):674-90. 10.1016/j.molcel.2012.05.021 PubMed 22681889 |
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 pFRT/TO/HIS/FLAG/HA-ALKBH5 6875 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pFRT/TO/HIS/FLAG/HA-ALKBH5
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6875)
AUTHORS Baltz AG, Munschauer M, Schwanhausser B, Vasile A, Murakawa Y,
Schueler M, Youngs N, Penfold-Brown D, Drew K, Milek M, Wyler E,
Bonneau R, Selbach M, Dieterich C, Landthaler M
TITLE The mRNA-Bound Proteome and Its Global Occupancy Profile on
Protein-Coding Transcripts.
JOURNAL Mol Cell. 2012 Jun 8;46(5):674-90.
PUBMED 22681889
REFERENCE 2 (bases 1 to 6875)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6875
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 67..108
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
primer_bind complement(146..163)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 152..376
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(509..528)
/label=F1ori-R
/note="F1 origin, reverse primer"
protein_bind 660..707
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
CDS 715..1737
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
"
polyA_signal 1867..1988
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1904..1923)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1958..1977
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(2037..2053)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2037..2053)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2050..2072)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2061..2077
/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(2085..2115)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2130..2151
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2268..2285)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2439..3027)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2519..2538)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3198..4058)
/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 3821..3840
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4059..4163)
/gene="bla"
/label=AmpR promoter
primer_bind complement(4238..4257)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 4429..4808
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 4809..5012
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 4963..4983
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
protein_bind 5014..5032
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 5035..5053
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 5063..5087
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 5187..5210
/codon_start=1
/product="8xHis affinity tag"
/label=8xHis
/translation="HHHHHHHH"
CDS 5214..5237
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 5238..5264
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
protein_bind 5280..5304
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(6690..6714)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 6767..6808
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
protein_bind complement(6865..14)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
ORIGIN
1 ttgtacaaag tggttgatat ccagcacagt ggcggccgct caagtctaga gggcccgcgg
61 ttcgaaggta agcctatccc taaccctctc ctcggtctcg attctacgct cgagtctaga
121 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
181 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
241 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
301 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
361 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
421 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
481 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
541 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
601 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtacctag
661 aagttcctat tccgaagttc ctattctcta gaaagtatag gaacttcctt ggccaaaaag
721 cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc
781 gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg
841 cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt
901 tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggagttc
961 agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg
1021 cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct
1081 gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa
1141 tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa
1201 actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt
1261 tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat
1321 gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg
1381 gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag
1441 cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg
1501 gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc
1561 gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact
1621 gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa
1681 gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaatagcac
1741 gtactacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
1801 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
1861 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
1921 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
1981 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
2041 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
2101 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
2161 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
2221 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
2281 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
2341 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
2401 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
2461 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
2521 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
2581 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
2641 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
2701 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
2761 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
2821 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
2881 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
2941 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
3001 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
3061 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
3121 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
3181 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
3241 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
3301 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
3361 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
3421 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
3481 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
3541 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
3601 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
3661 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
3721 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
3781 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
3841 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
3901 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
3961 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
4021 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
4081 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
4141 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
4201 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat
4261 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
4321 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
4381 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat
4441 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
4501 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
4561 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
4621 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
4681 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
4741 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
4801 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
4861 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg aaccaaaatc
4921 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
4981 gtgtacggtg ggaggtctat ataagcagag ctctccctat cagtgataga gatctcccta
5041 tcagtgatag agatcgtcga cgagctcgtt tagtgaaccg tcagatcgcc tggagacgcc
5101 atccacgctg ttttgacctc catagaagac accgggaccg atccagcctc cggactctag
5161 cgtttaaact taagcttggt accatgcacc accaccacca ccaccaccac atggactaca
5221 aggacgacga tgacaagtac ccttatgacg tgcccgatta cgctttgaac cagatatcaa
5281 caagtttgta caaaaaagca ggctccacca tgggaaccaa ttcagtcgac atggcggccg
5341 ccagcggcta cacggacctg cgtgagaagc tcaagtccat gacgtcccgg gacaactata
5401 aggcgggcag ccgggaggcc gccgccgctg ccgcagccgc cgctgccgcc gaaccttacc
5461 ctgtgtccgg ggccaagcgc aagtatcagg aggactcgga ccccgagcgc agcgactatg
5521 aggagcagca gctgcagaag gaggaggagg cgcgcaaggt gaagagcggc atccgccaga
5581 tgcgcctctt cagccaggac gagtgcgcca agatcgaggc ccgcattgac gaggtggtgt
5641 cccgcgctga gaagggcctg tacaacgagc acacggtgga ccgggcccca ctgcgcaaca
5701 agtacttctt cggcgaaggc tacacttacg gcgcccagct gcagaagcgc gggcccggcc
5761 aggagcgcct ctacccgccg ggcgacgtgg acgagatccc cgagtgggtg caccagctgg
5821 tgatccaaaa gctggtggag caccgcgtca tccccgaggg cttcgtcaac agcgccgtca
5881 tcaacgacta ccagcccggc ggctgcatcg tgtctcacgt ggaccccatc cacatcttcg
5941 agcgccccat cgtgtccgtg tccttcttta gcgactctgc gctgtgcttc ggctgcaagt
6001 tccagttcaa gcctattcgg gtgtcggaac cagtgctttc cctgccggtg cgcaggggaa
6061 gcgtgactgt gctcagtgga tatgctgctg atgaaatcac tcactgcata cggcctcagg
6121 acatcaagga gcgccgagca gtcatcatcc tcaggaagac aagattagat gcaccccggt
6181 tggaaacaaa gtccctgagc agctccgtgt taccacccag ctatgcttca gatcgcctgt
6241 caggaaacaa cagggaccct gctctgaaac ccaagcggtc ccaccgcaag gcagaccctg
6301 atgctgccca caggccacgg attctggaga tggacaagga agagaaccgg cgctcggtgc
6361 tgctgcccac acaccggcgg aggggtagct tcagctctga gaactactgg cgcaagtcat
6421 acgagtcctc agaggactgc tctgaggcag caggcagccc tgggaactct ggctcatcct
6481 tacgtagttg cccctccttt tgttttgagg gttttgtttt tgttcattgg ggggtttttg
6541 ttttttgttt tttgtttttt ttgattctat atatttttcc ttggttttgt tgcctgttag
6601 ggctgaagaa tagaattggc caggacctag gttctcatat tcttggtatt cctcctggat
6661 ggaaaggctg ttggcatcaa tagatctaga cccagctttc ttgtacaaag tggttgatat
6721 ccagcacagt ggcggccgct caagtctaga gggcccgcgg ttcgaaggta agcctatccc
6781 taaccctctc ctcggtctcg attctacgcg tacctgatcc gcggccgcat agataactga
6841 tccagtgtgc ctcgagatat ctagacccag ctttc
//
