| 质粒名称: | pcDNA3.1-MS2-Tet1-CD |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 83341 |
| 存储实验室: | Ronggui Hu |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1-hygro(+) |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达,CRISPR |
| 筛选标记: | 潮霉素 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | MS2-Tet1-CD |
| 物种来源: | Synthetic |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | -- |
| 克隆方法: | Gibson克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | BGH-rev |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3.1-MS2-Tet1-CD was a gift from Ronggui Hu (Addgene plasmid # 83341 ; http://n2t.net/addgene:83341 ; RRID:Addgene_83341) |
| 参考文献部分: | A CRISPR-based approach for targeted DNA demethylation. Xu X, Tao Y, Gao X, Zhang L, Li X, Zou W, Ruan K, Wang F, Xu GL, Hu R. Cell Discov. 2016 May 3;2:16009. doi: 10.1038/celldisc.2016.9. eCollection 2016. 10.1038/celldisc.2016.9 PubMed 27462456 |
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 pcDNA3.1-MS2-Tet1-CD 8104 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION To introduce MS2 coat protein fused Tet1-CD.
ACCESSION .
VERSION .
KEYWORDS pcDNA3.1-MS2-Tet1-CD
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8104)
AUTHORS Xu X, Tao Y, Gao X, Zhang L, Li X, Zou W, Ruan K, Wang F, Xu GL, Hu
R
TITLE A CRISPR-based approach for targeted DNA demethylation.
JOURNAL Cell Discov. 2016 May 3;2:16009. doi: 10.1038/celldisc.2016.9.
eCollection 2016.
PUBMED 27462456
REFERENCE 2 (bases 1 to 8104)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8104
/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"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 926..1312
/codon_start=1
/product="bacteriophage MS2 coat protein"
/label=MS2-N55K
/note="binds with high affinity to a specific stem-loop
structure in the viral RNA (Lim et al., 1994)"
/translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
CSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
VKAMQGLLKDGNPIPSAIAANSGIY"
CDS 1364..1384
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 3461..3484
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
primer_bind complement(3525..3542)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3531..3755
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 3801..4229
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3888..3907)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4098..4119
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4238..4258)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 4243..4573
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4424..4559
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4486..4505
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4622..5647
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
E"
polyA_signal 5777..5898
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5814..5833)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5868..5887
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5947..5963)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5947..5963)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5960..5982)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5971..5987
/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(5995..6025)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6040..6061
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6178..6195)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6349..6937)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6429..6448)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7108..7968)
/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 7731..7750
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7969..8073)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
901 gtttaaactt aagcttggta ccatggcttc taactttact cagttcgttc tcgtcgacaa
961 tggcggaact ggcgacgtga ctgtcgcccc aagcaacttc gctaacgggg tcgctgaatg
1021 gatcagctct aactcgcgtt cacaggctta caaagtaacc tgtagcgttc gtcagagctc
1081 tgcgcagaag cgcaaataca ccatcaaagt cgaggtgcct aaagtggcaa cccagactgt
1141 tggtggtgta gagcttcctg tagccgcatg gcgttcgtac ttaaatatgg aactaaccat
1201 tccaattttc gctacgaatt ccgactgcga gcttattgtt aaggcaatgc aaggtctcct
1261 aaaagatgga aacccgattc cctcagcaat cgcagcaaac tccggcatct acagcgctgg
1321 aggaggtgga agcggaggag gaggaagcgg aggaggaggt agccccaaga agaagagaaa
1381 ggtggaggcc agcggtggag gaagtggagg tggttcaggt ggaggatccg gcgaagctgc
1441 accctgtgac tgtgatggag gtacacaaaa agaaaaaggc ccatattata cacaccttgg
1501 ggcaggacca agtgtggctg ctgtcaggga gctcatggag actaggtttg gccagaaggg
1561 gaaggcaatc cggattgaga agatagtgtt cacggggaag gaagggaaga gctctcaggg
1621 ctgcccggtc gccaagtggg tgatcagaag aagtggtcct gaagagaagc ttatttgttt
1681 ggttcgtgag cgtgtagacc atcactgttc gacggctgtg atagttgtcc ttatcctgct
1741 gtgggaaggt atccctcgcc tgatggctga ccgcctgtac aaagagctca ctgagaactt
1801 gaggtcctac agcggacatc ccacagaccg aagatgtacc ctcaacaaaa agcgtacctg
1861 cacctgtcaa ggcatcgacc caaaaacctg cggagcgtcc ttctcctttg gctgttcgtg
1921 gagcatgtat ttcaacggct gtaagtttgg gaggagtgaa aaccccagaa aattcagact
1981 tgctccaaac taccccttac ataactacta taagagaatt actggaatga gttctgaagg
2041 aagtgacgtg aaaaccgggt ggatcattcc agaccgcaag accctcataa gcagagagga
2101 aaaacagctt gaaaagaatt tacaagaatt ggctacagta ttagctccac tttacaagca
2161 gatggctcca gttgcttatc aaaatcaggt ggaatatgaa gaagttgctg gagactgtcg
2221 acttggaaat gaagaggggc gtcctttctc tggtgtcacc tgttgcatgg atttttgtgc
2281 ccattctcac aaggacattc acaacatgca caacggaagc accgtggtgt gtacgttgat
2341 tcgagcagat ggccgtgaca caaattgtcc cgaggatgaa caactccacg tcctgccact
2401 ataccggctt gcagacactg atgaatttgg ctccgtggaa gggatgaagg ccaaaatcaa
2461 atctggggcc atccaagtca atgggccaac caggaagagg cgactacgtt ttactgagcc
2521 tgttcctcga tgtgggaaga gggccaaaat gaagcagaac cacaataaat caggttcaca
2581 caacactaag agcttttcat cagcctcatc tacttctcac ctagtgaaag acgaatctac
2641 agacttctgt cccctgcagg cttcctccgc agaaacatct acctgtacgt acagtaaaac
2701 agcctcaggt gggtttgcag aaacaagtag tattctccac tgcacaatgc cttctggagc
2761 acacagtggt gctaatgcag ctgctgggga atgtactgga acggtgcagc ctgccgaggt
2821 ggctgctcat cctcaccagt ctcttcccac agccgattct cccgttcatg ctgagcctct
2881 cactagtcca tctgagcagc taacttctaa ccagtcaaac cagcagctcc ctctcctcag
2941 caattctcag aaactggctt cctgtcaggt ggaagatgag cggcaccctg aagcggatga
3001 gcctcagcac cccgaggacg ataacttgcc tcaacttgat gagttctggt cagacagtga
3061 ggagatctac gccgatcctt cctttggtgg cgtggcgata gcacccattc acggctcggt
3121 gctcattgag tgcgctcgga aggagcttca tgctaccacc tctttgcgct cccccaaacg
3181 aggggtccct tttcgtgtgt cccttgtatt ctaccagcac aaaagcctaa acaagcctaa
3241 tcatggtttt gatatcaaca aaattaagtg taaatgcaaa aaagtaacga aaaaaaagcc
3301 cgcagaccgg gagtgtcctg atgtatcccc cgaagccaat ttatcacacc aaattccttc
3361 tcgagttgca tcaaccttaa cccgagacaa tgttgttacc gtgtccccat actctctcac
3421 tcatgttgcg ggaccctaca atcgttgggt cgcggccgca gactacaagg acgatgacga
3481 taagtaactc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
3541 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
3601 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
3661 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
3721 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
3781 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
3841 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
3901 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat
3961 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
4021 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
4081 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
4141 ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga
4201 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
4261 ggaaagtccc caggctcccc aggcaggcag aagtatgcaa agcatgcatc tcaattagtc
4321 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
4381 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
4441 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
4501 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
4561 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt
4621 gatgaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga
4681 cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga
4741 tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga
4801 tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat
4861 tggggaattc agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt
4921 gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggccatgga
4981 tgcgatcgct gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg
5041 aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta
5101 tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga
5161 gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg
5221 ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc
5281 gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc
5341 ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc
5401 gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga
5461 cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg
5521 agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg
5581 ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa
5641 ggaatagcac gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
5701 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
5761 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
5821 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
5881 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
5941 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
6001 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
6061 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
6121 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
6181 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
6241 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
6301 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
6361 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
6421 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
6481 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
6541 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
6601 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
6661 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
6721 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
6781 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
6841 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
6901 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
6961 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
7021 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
7081 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
7141 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
7201 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
7261 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
7321 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
7381 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
7441 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
7501 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
7561 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
7621 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
7681 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
7741 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
7801 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
7861 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
7921 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
7981 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
8041 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
8101 cgtc
//
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