| 质粒名称: | pET28-hTDG |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 70758 |
| 存储实验室: | Primo Schaer |
|
载体骨架基本信息 |
|
| 载体名称: | pET28c(+) |
| 载体出品公司: | Novagen |
| 空载体大小: | 5367bp |
| 完整质粒大小: | 6553bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | Kanamycin, 50 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | TDG、thymine DNA glycosylase |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1236 bp |
| GenBank编号 | NM_003211.4 |
| 启动子: | T7/lac operon |
| 标签/融合蛋白: | 6-HIS (N terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NheI (not destroyed) |
| 3’克隆位点: | SalI (not destroyed) |
| 5’测序引物 | pBRrevBam |
| 3’测序引物 | T7 terminal |
|
文献引用方法 |
|
| 材料和方法部分: | pET28-hTDG was a gift from Primo Schaer (Addgene plasmid # 70758 ; http://n2t.net/addgene:70758 ; RRID:Addgene_70758) |
| 参考文献部分: | 3CAPS - a structural AP-site analogue as a tool to investigate DNA base excision repair. Schuermann D, Scheidegger SP, Weber AR, Bjoras M, Leumann CJ, Schar P. Nucleic Acids Res. 2016 Jan 4. pii: gkv1520. 10.1093/nar/gkv1520 PubMed 26733580 |
买家导航
pET28-hTDG载体质粒基本信息
pET28-hTDG质粒图谱载体图谱和pET28-hTDG载体序列质粒序列多克隆位点信息
pET28-hTDG质粒载体简介
pET28-hTDG质粒序列载体序列
LOCUS pET28-hTDG 6553 bp DNA circular SYN 14-APR-2025
DEFINITION bacterial expression of human TDG.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6553)
AUTHORS Schuermann D, Scheidegger SP, Weber AR, Bjoras M, Leumann CJ, Schar
P
TITLE 3CAPS - a structural AP-site analogue as a tool to investigate DNA
base excision repair.
JOURNAL Nucleic Acids Res. 2016 Jan 4. pii: gkv1520.
PUBMED 26733580
REFERENCE 2 (bases 1 to 6553)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res. 2016 Jan 4. pii: gkv1520."
COMMENT Sequence Label: pET28-hTDG
FEATURES Location/Qualifiers
source 1..6553
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter complement(4..81)
/gene="lacI"
/label=lacI promoter
primer_bind 287..306
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 390..409
/label=T7
/note="T7 promoter, forward primer"
promoter 390..408
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 409..433
/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)."
RBS 448..470
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 489..506
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 516..533
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS 1803..1820
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(1873..1891)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 1887..1934
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 1971..2426
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2058..2077)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2268..2289
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(2519..3334)
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
CDS complement(2519..3334)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 3242..3261
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 3326..3345
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 3456..4044
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3945..3964
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4198..4215
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 4230..4372
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(4298..4317)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4458..4480
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(4474..4665)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(join(5474..6553,1..3))
/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 6518..6537
/label=LacI-R
/note="LacI, reverse primer"
ORIGIN
1 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
61 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
121 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
181 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
241 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
301 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
361 agaggatcga gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc
421 ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga tataccatgg
481 gcagcagcca tcatcatcat catcacagca gcggcctggt gccgcgcggc agccatatgg
541 ctagcgaagc ggagaacgcg ggcagctatt cccttcagca agctcaagct ttttatacgt
601 ttccatttca acaactgatg gctgaagctc ctaatatggc agttgtgaat gaacagcaaa
661 tgccagaaga agttccagcc ccagctcctg ctcaggaacc agtgcaagag gctccaaaag
721 gaagaaaaag aaaacccaga acaacagaac caaaacaacc agtggaaccc aaaaaacctg
781 ttgagtcaaa aaaatctggc aagtctgcaa aatcaaaaga aaaacaagaa aaaattacag
841 acacatttaa agtaaaaaga aaagtagacc gttttaatgg tgtttcagaa gctgaacttc
901 tgaccaagac tctccccgat attttgacct tcaatctgga cattgtcatt attggcataa
961 acccgggact aatggctgct tacaaagggc atcattaccc tggacctgga aaccattttt
1021 ggaagtgttt gtttatgtca gggctcagtg aggtccagct gaaccatatg gatgatcaca
1081 ctctaccagg gaagtatggt attggattta ccaacatggt ggaaaggacc acgcccggca
1141 gcaaagatct ctccagtaaa gaatttcgtg aaggaggacg tattctagta cagaaattac
1201 agaaatatca gccacgaata gcagtgttta atggaaaatg tatttatgaa atttttagta
1261 aagaagtttt tggagtaaag gttaagaact tggaatttgg gcttcagccc cataagattc
1321 cagacacaga aactctctgc tatgttatgc catcatccag tgcaagatgt gctcagtttc
1381 ctcgagccca agacaaagtt cattactaca taaaactgaa ggacttaaga gatcagttga
1441 aaggcattga acgaaatatg gacgttcaag aggtgcaata tacatttgac ctacagcttg
1501 cccaagagga tgcaaagaag atggctgtta aggaagaaaa atatgatcca ggttatgagg
1561 cagcatatgg tggtgcttac ggagaaaatc catgcagcag tgaaccttgt ggcttctctt
1621 caaatgggct aattgagagc gtggagttaa gaggagaatc agctttcagt ggcattccta
1681 atgggcagtg gatgacccag tcatttacag accaaattcc ttcctttagt aatcactgtg
1741 gaacacaaga acaggaagaa gaaagccatg cttaagtcga caagcttgcg gccgcactcg
1801 agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt
1861 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
1921 tgaggggttt tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg
1981 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
2041 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
2101 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
2161 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
2221 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
2281 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
2341 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
2401 taacaaaata ttaacgttta caatttcagg tggcactttt cggggaaatg tgcgcggaac
2461 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga attaattctt
2521 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
2581 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
2641 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
2701 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
2761 aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc
2821 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
2881 cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat
2941 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
3001 cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat
3061 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
3121 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
3181 actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat
3241 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
3301 tagagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt
3361 aagcagacag ttttattgtt catgaccaaa atcccttaac gtgagttttc gttccactga
3421 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
3481 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
3541 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
3601 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
3661 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
3721 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
3781 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
3841 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
3901 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
3961 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
4021 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
4081 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
4141 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
4201 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
4261 tgcggtattt cacaccgcat atatggtgca ctctcagtac aatctgctct gatgccgcat
4321 agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca
4381 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
4441 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
4501 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
4561 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
4621 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta
4681 agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat
4741 acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc
4801 ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa
4861 tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat
4921 aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac
4981 cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc
5041 gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct
5101 caacgacagg agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg
5161 cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa
5221 gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc
5281 gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac
5341 agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
5401 gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt
5461 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
5521 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
5581 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
5641 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
5701 acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg
5761 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
5821 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
5881 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
5941 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
6001 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
6061 cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
6121 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
6181 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
6241 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
6301 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
6361 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
6421 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
6481 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
6541 acgttactgg ttt
//
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