| 质粒名称: | pCDF-His-mTET1CD |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 81053 |
| 存储实验室: | Primo Schaer |
|
载体骨架基本信息 |
|
| 载体名称: | pCDF-Duet-1 |
| 载体出品公司: | Novagen |
| 载体抗性: | 链霉素 |
| 空载体大小: | 3780 bp |
| 完整质粒大小: | 5590 bp |
| 载体类型: | 细菌表达 |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | TET1 |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 2082 bp |
| 基因突变: | catalytic domain amino acid 1367-2057 |
| GenBank编号 | NM_001253857.1 |
| 启动子 | T7 |
| 标签/融合蛋白: | 6HIS（主干上的N终端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NotI (未破坏) |
| 3' 克隆位点: | PacI (未破坏) |
| 5' 测序引物 | GGATCTCGACGCTCTCCCT |
| 3' 测序引物 | T7_terminal_primer |
|
文献引用方法 |
|
| 材料和方法部分: | pCDF-His-mTET1CD was a gift from Primo Schaer (Addgene plasmid # 81053 ; http://n2t.net/addgene:81053 ; RRID:Addgene_81053) |
| 参考文献部分: | Biochemical reconstitution of TET1-TDG-BER-dependent active DNA demethylation reveals a highly coordinated mechanism. Weber AR, Krawczyk C, Robertson AB, Kusnierczyk A, Vagbo CB, Schuermann D, Klungland A, Schar P. Nat Commun. 2016 Mar 2;7:10806. doi: 10.1038/ncomms10806. 10.1038/ncomms10806 PubMed 26932196 |
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 pCDF-His-mTET1CD 5590 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION bacterial expression of the catalytic domain of murine TET1.
ACCESSION .
VERSION .
KEYWORDS pCDF-His-mTET1CD
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5590)
AUTHORS Weber AR, Krawczyk C, Robertson AB, Kusnierczyk A, Vagbo CB,
Schuermann D, Klungland A, Schar P
TITLE Biochemical reconstitution of TET1-TDG-BER-dependent active DNA
demethylation reveals a highly coordinated mechanism.
JOURNAL Nat Commun. 2016 Mar 2;7:10806. doi: 10.1038/ncomms10806.
PUBMED 26932196
REFERENCE 2 (bases 1 to 5590)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5590
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 115..134
/label=T7
/note="T7 promoter, forward primer"
promoter 115..133
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 134..158
/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)."
CDS 214..231
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(2388..2406)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 2402..2449
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(2620..3411)
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
promoter complement(3412..3503)
/gene="bla"
/label=AmpR promoter
rep_origin complement(3551..4289)
/direction=LEFT
/label=CloDF13 ori
/note="Plasmids containing the CloDF13 (CDF) origin of
replication can be propagated in E. coli cells that contain
additional plasmids with compatible origins."
CDS complement(4499..5581)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind 5543..5562
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(5582..69)
/gene="lacI"
/label=lacI promoter
ORIGIN
1 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
61 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aaattaatac
121 gactcactat aggggaattg tgagcggata acaattcccc tgtagaaata attttgttta
181 actttaataa ggagatatac catgggcagc agccatcacc atcatcacca cagccaggat
241 ccgaattcga gctcggcgcg cctgcaggtc gacaagcttg cggccgcaga agctgcaccc
301 tgtgactgtg atggaggtac acaaaaagaa aaaggcccat attatacaca ccttggggca
361 ggaccaagtg tggctgctgt cagggagctc atggagacta ggtttggcca gaaggggaag
421 gcaatccgga ttgagaagat agtgttcacg gggaaggaag ggaagagctc tcagggctgc
481 ccggtcgcca agtgggtgat cagaagaagt ggtcctgaag agaagcttat ttgtttggtt
541 cgtgagcgtg tagaccatca ctgttcgacg gctgtgatag ttgtccttat cctgctgtgg
601 gaaggtatcc ctcgcctgat ggctgaccgc ctgtacaaag agctcactga gaacttgagg
661 tcctacagcg gacatcccac agaccgaaga tgtaccctca acaaaaagcg tacctgcacc
721 tgtcaaggca tcgacccaaa aacctgcgga gcgtccttct cctttggctg ttcgtggagc
781 atgtatttca acggctgtaa gtttgggagg agtgaaaacc ccagaaaatt cagacttgct
841 ccaaactacc ccttacataa ctactataag agaattactg gaatgagttc tgaaggaagt
901 gacgtgaaaa ccgggtggat cattccagac cgcaagaccc tcataagcag agaggaaaaa
961 cagcttgaaa agaatttaca agaattggct acagtattag ctccacttta caagcagatg
1021 gctccagttg cttatcaaaa tcaggtggaa tatgaagaag ttgctggaga ctgtcgactt
1081 ggaaatgaag aggggcgtcc tttctctggt gtcacctgtt gcatggattt ttgtgcccat
1141 tctcacaagg acattcacaa catgcacaac ggaagcaccg tggtgtgtac gttgattcga
1201 gcagatggcc gtgacacaaa ttgtcccgag gatgaacaac tccacgtcct gccactatac
1261 cggcttgcag acactgatga atttggctcc gtggaaggga tgaaggccaa aatcaaatct
1321 ggggccatcc aagtcaatgg gccaaccagg aagaggcgac tacgttttac tgagcctgtt
1381 cctcgatgtg ggaagagggc caaaatgaag cagaaccaca ataaatcagg tactgctggt
1441 ctcagaagga agaggatctc tgccagccca aagggagccc caggttcaca caacactaag
1501 agcttttcat cagcctcatc tacttctcac ctagtgaaag acgaatctac agacttctgt
1561 cccctgcagg cttcctccgc agaaacatct acctgtacgt acagtaaaac agcctcaggt
1621 gggtttgcag aaacaagtag tattctccac tgcacaatgc cttctggagc acacagtggt
1681 gctaatgcag ctgctgggga atgtactgga acggtgcagc ctgccgaggt ggctgctcat
1741 cctcaccagt ctcttcccac agccgattct cccgttcatg ctgagcctct cactagtcca
1801 tctgagcagc taacttctaa ccagtcaaac cagcagctcc ctctcctcag caattctcag
1861 aaactggctt cctgtcaggt ggaagatgag cggcaccctg aagcggatga gcctcagcac
1921 cccgaggacg ataacttgcc tcaacttgat gaattctggt cagacagtga ggagatctac
1981 gccgatcctt cctttggtgg cgtggcgata gcacccattc acggctcggt gctcattgag
2041 tgcgctcgga aggagcttca tgctaccacc tctttgcgct cccccaaacg aggggtccct
2101 tttcgtgtgt cccttgtatt ctaccagcac aaaagcctaa acaagcctaa tcatggtttt
2161 gatatcaaca aaattaagtg taaatgcaaa aaagtaacga aaaaaaagcc cgcagaccgg
2221 gagtgtcctg atgtatcccc cgaagccaat ttatcacacc aaattccttc tcgagttgca
2281 tcaaccttaa cccgagacaa tgttgttacc gtgtccccat actctctcac tcatgttgcg
2341 ggaccctaca atcgttgggt ctaattaatt aacctaggct gctgccaccg ctgagcaata
2401 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaacctca
2461 ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca
2521 atagacataa gcggctattt aacgaccctg ccctgaaccg acgaccgggt catcgtggcc
2581 ggatcttgcg gcccctcggc ttgaacgaat tgttagacat tatttgccga ctaccttggt
2641 gatctcgcct ttcacgtagt ggacaaattc ttccaactga tctgcgcgcg aggccaagcg
2701 atcttcttct tgtccaagat aagcctgtct agcttcaagt atgacgggct gatactgggc
2761 cggcaggcgc tccattgccc agtcggcagc gacatccttc ggcgcgattt tgccggttac
2821 tgcgctgtac caaatgcggg acaacgtaag cactacattt cgctcatcgc cagcccagtc
2881 gggcggcgag ttccatagcg ttaaggtttc atttagcgcc tcaaatagat cctgttcagg
2941 aaccggatca aagagttcct ccgccgctgg acctaccaag gcaacgctat gttctcttgc
3001 ttttgtcagc aagatagcca gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag
3061 aatgtcattg cgctgccatt ctccaaattg cagttcgcgc ttagctggat aacgccacgg
3121 aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg cggagaatct cgctctctcc
3181 aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag
3241 ccttacggtc accgtaacca gcaaatcaat atcactgtgt ggcttcaggc cgccatccac
3301 tgcggagccg tacaaatgta cggccagcaa cgtcggttcg agatggcgct cgatgacgcc
3361 aactacctct gatagttgag tcgatacttc ggcgatcacc gcttccctca tactcttcct
3421 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
3481 atgtatttag aaaaataaac aaatagctag ctcactcggt cgctacgctc cgggcgtgag
3541 actgcggcgg gcgctgcgga cacatacaaa gttacccaca gattccgtgg ataagcaggg
3601 gactaacatg tgaggcaaaa cagcagggcc gcgccggtgg cgtttttcca taggctccgc
3661 cctcctgcca gagttcacat aaacagacgc ttttccggtg catctgtggg agccgtgagg
3721 ctcaaccatg aatctgacag tacgggcgaa acccgacagg acttaaagat ccccaccgtt
3781 tccggcgggt cgctccctct tgcgctctcc tgttccgacc ctgccgttta ccggatacct
3841 gttccgcctt tctcccttac gggaagtgtg gcgctttctc atagctcaca cactggtatc
3901 tcggctcggt gtaggtcgtt cgctccaagc tgggctgtaa gcaagaactc cccgttcagc
3961 ccgactgctg cgccttatcc ggtaactgtt cacttgagtc caacccggaa aagcacggta
4021 aaacgccact ggcagcagcc attggtaact gggagttcgc agaggatttg tttagctaaa
4081 cacgcggttg ctcttgaagt gtgcgccaaa gtccggctac actggaagga cagatttggt
4141 tgctgtgctc tgcgaaagcc agttaccacg gttaagcagt tccccaactg acttaacctt
4201 cgatcaaacc acctccccag gtggtttttt cgtttacagg gcaaaagatt acgcgcagaa
4261 aaaaaggatc tcaagaagat cctttgatct tttctactga accgctctag atttcagtgc
4321 aatttatctc ttcaaatgta gcacctgaag tcagccccat acgatataag ttgtaattct
4381 catgttagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
4441 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
4501 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
4561 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
4621 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
4681 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
4741 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc
4801 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
4861 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
4921 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
4981 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
5041 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
5101 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
5161 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
5221 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
5281 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
5341 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
5401 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
5461 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
5521 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
5581 cattcaccac
//
