| 质粒名称: | pJFA334E9 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 49237 |
| 存储实验室: | Keith Joung |
|
载体骨架基本信息 |
|
| 载体名称: | -- |
| 载体出品公司: | Joung lab |
| 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: | TALEN；TALE-TET1表达载体 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | TET1 tet methylcytosine dioxygenase 1 |
| 物种来源: | H. sapiens (human), Synthetic |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | EF1a |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | OMM520 |
| 3' 测序引物 | OJA199 |
|
文献引用方法 |
|
| 材料和方法部分: | pJFA334E9 was a gift from Keith Joung (Addgene plasmid # 49237 ; http://n2t.net/addgene:49237 ; RRID:Addgene_49237) |
| 参考文献部分: | Targeted DNA demethylation and activation of endogenous genes using programmable TALE-TET1 fusion proteins. Maeder ML, Angstman JF, Richardson ME, Linder SJ, Cascio VM, Tsai SQ, Ho QH, Sander JD, Reyon D, Bernstein BE, Costello JF, Wilkinson MF, Joung JK. Nat Biotechnol. 2013 Oct 9. doi: 10.1038/nbt.2726. 10.1038/nbt.2726 PubMed 24108092 |
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 pJFA334E9 8121 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION TET1-catalytic domain (CD) expression vector for TALE-TET1 fusion
protein to induce targeted demethylation in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pJFA334E9
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8121)
AUTHORS Maeder ML, Angstman JF, Richardson ME, Linder SJ, Cascio VM, Tsai
SQ, Ho QH, Sander JD, Reyon D, Bernstein BE, Costello JF, Wilkinson
MF, Joung JK
TITLE Targeted DNA demethylation and activation of endogenous genes using
programmable TALE-TET1 fusion proteins.
JOURNAL Nat Biotechnol. 2013 Oct 9. doi: 10.1038/nbt.2726.
PUBMED 24108092
REFERENCE 2 (bases 1 to 8121)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8121
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 331..350
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(569..673)
/gene="bla"
/label=AmpR promoter
primer_bind complement(752..771)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 927..2105
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 1158..2096
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 2053..2073
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
regulatory 2113..2122
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2125..2145
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(5161..5180)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5215..5234
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5271..5291)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 5276..5605
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 5456..5591
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5518..5537
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 5615..6214
/codon_start=1
/gene="pac from Streptomyces"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
primer_bind complement(5615..5634)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 6111..6131
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
CDS 6276..6317
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
CDS 6327..6344
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(6367..6384)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 6373..6597
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(6668..6684)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6668..6684)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6681..6703)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6692..6708
/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(6716..6746)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6761..6782
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6899..6916)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7070..7658)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7150..7169)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7829..568)
/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"
ORIGIN
1 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
61 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
121 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
181 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
241 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
301 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
361 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
421 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
481 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
541 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
601 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
661 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
721 gatctcccga tcccctaggg tcgactctca gtacaatctg ctctgatgcc gcatagttaa
781 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
841 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
901 gttttgcgct gcttcgctag tcgtgaggct ccggtgcccg tcagtgggca gagcgcacat
961 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa
1021 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
1081 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
1141 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
1201 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc
1261 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
1321 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
1381 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
1441 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
1501 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
1561 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
1621 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
1681 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
1741 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
1801 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
1861 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
1921 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
1981 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
2041 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
2101 cgtgaagcgg ccgccaccat ggcccccaag aagaagagga aggtgggcat tcaccgcggg
2161 gtacctatgg tggacttgag gacactcggt tattcgcaac agcaacagga gaaaatcaag
2221 cctaaggtca ggagcaccgt cgcgcaacac cacgaggcgc ttgtggggca tggcttcact
2281 catgcgcata ttgtcgcgct ttcacagcac cctgcggcgc ttgggacggt ggctgtcaaa
2341 taccaagata tgattgcggc cctgcccgaa gccacgcacg aggcaattgt aggggtcggt
2401 aaacagtggt cgggagcgcg agcacttgag gcgctgctga ctgtggcggg tgagcttagg
2461 gggcctccgc tccagctcga caccgggcag ctgctgaaga tcgcgaagag agggggagta
2521 acagcggtag aggcagtgca cgcctggcgc aatgcgctca ccggggcccc cttgaacaga
2581 gacgatctag agctagacgt ctcgctgaca cccgaacagg tggtcgccat tgcttcccac
2641 gacggaggac ggccagcctt ggagagcatt gtggcccagc tgtccaggcc ggaccctgcc
2701 ctggccgcgt taaccaacga ccacctcgtc gccttggcct gcctcggcgg acgtcctgcg
2761 ctggatgcag tgaaaaaggg attgccgcac gcgccggcct tgatcaaaag aaccaatcgc
2821 cgtattcccg aacgcacatc ccatcgcgtt gccgaccacg cgcaagtggt tcgcgtgctg
2881 ggttttttcc agtgccactc ccacccagcg caagcatttg atgacgccat gacgcagttc
2941 gggatgagcg gaggaggatc cctgcccacc tgcagctgtc ttgatcgagt tatacaaaaa
3001 gacaaaggcc catattatac acaccttggg gcaggaccaa gtgttgctgc tgtcagggaa
3061 atcatggaga ataggtatgg tcaaaaagga aacgcaataa ggatagaaat agtagtgtac
3121 accggtaaag aagggaaaag ctctcatggg tgtccaattg ctaagtgggt tttaagaaga
3181 agcagtgatg aagaaaaagt tctttgtttg gtccggcagc gtacaggcca ccactgtcca
3241 actgctgtga tggtggtgct catcatggtg tgggatggca tccctcttcc aatggccgac
3301 cggctataca cagagctcac agagaatcta aagtcataca atgggcaccc taccgacaga
3361 agatgcaccc tcaatgaaaa tcgtacctgt acatgtcaag gaattgatcc agagacttgt
3421 ggagcttcat tctcttttgg ctgttcatgg agtatgtact ttaatggctg taagtttggt
3481 agaagcccaa gccccagaag atttagaatt gatccaagct ctcccttaca tgaaaaaaac
3541 cttgaagata acttacagag tttggctaca cgattagctc caatttataa gcagtatgct
3601 ccagtagctt accaaaatca ggtggaatat gaaaatgttg cccgagaatg tcggcttggc
3661 agcaaggaag gtcgaccctt ctctggggtc actgcttgcc tggacttctg tgctcatccc
3721 cacagggaca ttcacaacat gaataatgga agcactgtgg tttgtacctt aactcgagaa
3781 gataaccgct ctttgggtgt tattcctcaa gatgagcagc tccatgtgct acctctttat
3841 aagctttcag acacagatga gtttggctcc aaggaaggaa tggaagccaa gatcaaatct
3901 ggggccatcg aggtcctggc accccgccgc aaaaaaagaa cgtgtttcac tcagcctgtt
3961 ccccgttctg gaaagaagag ggctgcgatg atgacagagg ttcttgcaca taagataagg
4021 gcagtggaaa agaaacctat tccccgaatc aagcggaaga ataactcaac aacaacaaac
4081 aacagtaagc cttcgtcact gccaacctta gggagtaaca ctgagaccgt gcaacctgaa
4141 gtaaaaagtg aaaccgaacc ccattttatc ttaaaaagtt cagacaacac taaaacttat
4201 tcgctgatgc catccgctcc tcacccagtg aaagaggcat ctccaggctt ctcctggtcc
4261 ccgaagactg cttcagccac accagctcca ctgaagaatg acgcaacagc ctcatgcggg
4321 ttttcagaaa gaagcagcac tccccactgt acgatgcctt cgggaagact cagtggtgcc
4381 aatgctgcag ctgctgatgg ccctggcatt tcacagcttg gcgaagtggc tcctctcccc
4441 accctgtctg ctcctgtgat ggagcccctc attaattctg agccttccac tggtgtgact
4501 gagccgctaa cgcctcatca gccaaaccac cagccctcct tcctcacctc tcctcaagac
4561 cttgcctctt ctccaatgga agaagatgag cagcattctg aagcagatga gcctccatca
4621 gacgaacccc tatctgatga ccccctgtca cctgctgagg agaaattgcc ccacattgat
4681 gagtattggt cagacagtga gcacatcttt ttggatgcaa atattggtgg ggtggccatc
4741 gcacctgctc acggctcggt tttgattgag tgtgcccggc gagagctgca cgctaccact
4801 cctgttgagc accccaaccg taatcatcca acccgcctct cccttgtctt ttaccagcac
4861 aaaaacctaa ataagcccca acatggtttt gaactaaaca agattaagtt tgaggctaaa
4921 gaagctaaga ataagaaaat gaaggcctca gagcaaaaag accaggcagc taatgaaggt
4981 ccagaacagt cctctgaagt aaatgaattg aaccaaattc cttctcataa agcattaaca
5041 ttaacccatg acaatgttgt caccgtgtcc ccttatgctc tcacacacgt tgcggggccc
5101 tataaccatt gggtctgatt aattaattgt ttattgcagc ttataatggt tacaaataaa
5161 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
5221 tgtccaaact catcaatgta tcttatgaat tccgattaag tcgaccctgt ggaatgtgtg
5281 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
5341 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
5401 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
5461 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
5521 ttatgcagag gccgaggccg cctcggcttc tgagctattc cagaagtagt gaggaggctt
5581 ttttggaggc ctaggctttt gcaaaaagct taccatgacc gagtacaagc ccacggtgcg
5641 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
5701 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
5761 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
5821 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
5881 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
5941 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
6001 cgtgtcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
6061 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
6121 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gagtgcccga aggaccgcgc
6181 gacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc
6241 gaccgaaagg agcgcacgac cccatgcatc tcgaaggtaa gcctatccct aaccctctcc
6301 tcggtctcga ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct
6361 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
6421 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
6481 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
6541 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tccatggctt
6601 ctgaggcgga aagaaccagc tggggctcga taccgtcgac ctctagctag agcttggcgt
6661 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
6721 tacgagccgg aagcataaag tgtaaagcct agggtgccta atgagtgagc taactcacat
6781 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
6841 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
6901 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
6961 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
7021 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
7081 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
7141 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
7201 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
7261 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
7321 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
7381 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
7441 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
7501 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
7561 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
7621 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
7681 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
7741 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
7801 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
7861 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
7921 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
7981 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
8041 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
8101 gtagttcgcc agttaatagt t
//
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