| 质粒名称: | pFRT_TO_FlagHA_TIAL1 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 106090 |
| 存储实验室: | Thomas Tuschl |
|
载体骨架基本信息 |
|
| 载体名称: | pFRT_TO_FlagHA_dest |
| 完整质粒大小: | 6464 bp |
| 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: | TIAL1 |
| 物种来源: | H. sapiens (human) |
| 标签/融合蛋白: | FlagHA (N terminal on backbone) |
| 克隆方法: | Gateway Cloning |
| 5’测序引物 | CMV-F |
| 3’测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pFRT_TO_FlagHA_TIAL1 was a gift from Thomas Tuschl (Addgene plasmid # 106090 ; http://n2t.net/addgene:106090 ; RRID:Addgene_106090) |
| 参考文献部分: | The TIA1 RNA-Binding Protein Family Regulates EIF2AK2-Mediated Stress Response and Cell Cycle Progression. Meyer C, Garzia A, Mazzola M, Gerstberger S, Molina H, Tuschl T. Mol Cell. 2018 Feb 15;69(4):622-635.e6. doi: 10.1016/j.molcel.2018.01.011. 10.1016/j.molcel.2018.01.011 PubMed 29429924 |
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_FlagHA_TIAL1 6464 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION gene expression of FlagHA-TIAL1.
ACCESSION .
VERSION .
KEYWORDS pFRT_TO_FlagHA_TIAL1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6464)
AUTHORS Meyer C, Garzia A, Mazzola M, Gerstberger S, Molina H, Tuschl T
TITLE The TIA1 RNA-Binding Protein Family Regulates EIF2AK2-Mediated
Stress Response and Cell Cycle Progression.
JOURNAL Mol Cell. 2018 Feb 15;69(4):622-635.e6. doi:
10.1016/j.molcel.2018.01.011.
PUBMED 29429924
REFERENCE 2 (bases 1 to 6464)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6464
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
protein_bind 820..838
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 841..859
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 869..893
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 993..1016
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 1017..1043
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
protein_bind 1059..1083
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(2218..2242)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 2295..2336
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
primer_bind complement(2416..2433)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2422..2646
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(2779..2798)
/label=F1ori-R
/note="F1 origin, reverse primer"
protein_bind 2930..2977
/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 2985..4007
/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 4137..4258
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4174..4193)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4228..4247
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4307..4323)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4307..4323)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4320..4342)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4331..4347
/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(4355..4385)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4400..4421
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4538..4555)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4709..5297)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4789..4808)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5468..6328)
/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 6091..6110
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6329..6433)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
961 ctctagcgtt taaacttaag cttggtacca tggactacaa ggacgacgat gacaagtacc
1021 cttatgacgt gcccgattac gctttgaacc agatatcaac aagtttgtac aaaaaagcag
1081 gcttcatgat ggaagacgac gggcagcccc ggactctata cgtaggtaac ctttccagag
1141 atgtgacaga agtccttata cttcagttgt tcagtcagat tggaccctgt aaaagctgta
1201 aaatgataac agagcataca agcaatgacc catattgctt tgtggaattt tatgaacaca
1261 gagatgcagc tgctgcatta gctgctatga atgggagaaa aattttggga aaggaggtca
1321 aagtaaactg ggcaaccaca ccaagtagcc agaaaaaaga tacttccaat cacttccatg
1381 tgtttgttgg ggatttgagt ccagaaatta caacagaaga tatcaaatca gcatttgccc
1441 cctttggtaa aatatcggat gcccgggtag ttaaagacat ggcaactgga aaatccaaag
1501 gctatggttt tgtatctttt tataacaaac tggatgcaga aaatgcgatt gtgcatatgg
1561 gcggtcagtg gttgggtggt cgtcaaatcc gaaccaattg ggccactcgt aaaccacctg
1621 cacctaaaag tacacaagaa aacaacacta agcagttgag atttgaagat gtagtaaacc
1681 agtcaagtcc aaaaaattgt actgtgtact gtggaggaat tgcgtctggg ttaacagatc
1741 agcttatgag acagacattc tcaccatttg gacaaattat ggaaataaga gttttcccag
1801 aaaagggcta ttcatttgtc agattttcaa cccatgaaag tgcagcccat gccattgttt
1861 cggtgaacgg tactacgatt gaaggacatg tggttaaatg ctattggggt aaagaatctc
1921 ctgatatgac taaaaacttc caacaggttg actatagtca atggggccaa tggagccaag
1981 tgtatggaaa cccacaacag tatggacagt atatggcaaa tgggtggcaa gtaccgcctt
2041 atggagtata cgggcaacca tggaatcaac aaggatttgg agtagatcaa tcaccttctg
2101 ctgcttggat gggtggattt ggtgctcagc ctccccaagg acaagctcct ccccctgtaa
2161 tacctcctcc taaccaagcc ggatatggta tggcaagtta ccaaacacag tgataagacc
2221 cagctttctt gtacaaagtg gttgatatcc agcacagtgg cggccgctca agtctagagg
2281 gcccgcggtt cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta
2341 cctgatccgc ggccgcatag ataactgatc cagtgtgcct cgagtctaga gggcccgttt
2401 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
2461 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
2521 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
2581 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
2641 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
2701 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
2761 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
2821 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
2881 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtacctag aagttcctat
2941 tccgaagttc ctattctcta gaaagtatag gaacttcctt ggccaaaaag cctgaactca
3001 ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc
3061 agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg
3121 tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact
3181 ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggaattc agcgagagcc
3241 tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg
3301 aactgcccgc tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc
3361 ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat
3421 ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg
3481 acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg
3541 actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg
3601 acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat
3661 acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc
3721 gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg gcgtatatgc
3781 tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag
3841 cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta
3901 cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg
3961 atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaatagcac gtactacgag
4021 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
4081 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
4141 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
4201 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
4261 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
4321 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
4381 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
4441 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
4501 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
4561 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
4621 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
4681 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
4741 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
4801 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
4861 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
4921 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
4981 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
5041 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
5101 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
5161 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
5221 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
5281 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
5341 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
5401 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
5461 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
5521 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
5581 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
5641 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
5701 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
5761 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
5821 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
5881 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
5941 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
6001 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
6061 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
6121 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
6181 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
6241 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
6301 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
6361 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
6421 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
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