| 质粒名称: | pSANG10-3F-BG4 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 55756 |
| 存储实验室: | Shankar Balasubramanian |
|
载体骨架基本信息 |
|
| 载体名称: | pSANG10-3F |
| 载体出品公司: | John McCafferty |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 5438 bp |
| 完整质粒大小: | 6197 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 30 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | BG4,BG4 anti-G-quadruplex scFv antibody |
| 物种来源: | human phage display antibody from Sanger library |
| 插入大小: | 759 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | T7 |
| 标签/融合蛋白: |
6xHis（主干上的C终端）
tri-FLAG（主干上的C终端）
pelB引线（主干上的N端子）
|
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | Ncol(未破坏) |
| 3' 克隆位点: | Notl (未破坏) |
| 5' 测序引物 | T7 promoter |
| 3' 测序引物 | T7 terminal primer |
|
文献引用方法 |
|
| 材料和方法部分: | pSANG10-3F-BG4 was a gift from Shankar Balasubramanian (Addgene plasmid # 55756 ; http://n2t.net/addgene:55756 ; RRID:Addgene_55756) |
| 参考文献部分: | Quantitative visualization of DNA G-quadruplex structures in human cells. Biffi G, Tannahill D, McCafferty J, Balasubramanian S. Nat Chem. 2013 Mar;5(3):182-6. doi: 10.1038/nchem.1548. Epub 2013 Jan 20. 10.1038/nchem.1548 PubMed 23422559 |
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 pSANG10-3F-BG4 6169 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses BG4 scFv anti-G-quadruplex antibody in bacteria.
ACCESSION .
VERSION .
KEYWORDS pSANG10-3F-BG4
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6169)
AUTHORS Biffi G, Tannahill D, McCafferty J, Balasubramanian S
TITLE Quantitative visualization of DNA G-quadruplex structures in human
cells.
JOURNAL Nat Chem. 2013 Mar;5(3):182-6. doi: 10.1038/nchem.1548. Epub 2013
Jan 20.
PUBMED 23422559
REFERENCE 2 (bases 1 to 6169)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6169
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 1..140
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(66..85)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 226..248
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(242..433)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(1242..2324)
/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 2286..2305
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(2325..2402)
/gene="lacI"
/label=lacI promoter
primer_bind 2608..2627
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 2711..2730
/label=T7
/note="T7 promoter, forward primer"
promoter 2711..2729
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 2730..2754
/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 2769..2791
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 2799..2864
/codon_start=1
/gene="pelB (fragment)"
/product="leader peptide for secretion"
/label=pelB signal sequence
/translation="MKYLLPTAAAGLLLLAAQPAMA"
CDS 3630..3647
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 3654..3719
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 3744..3761
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(3814..3832)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 3828..3875
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 3912..4367
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3999..4018)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4209..4230
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(4459..5274)
/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 5182..5201
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 5266..5285
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 5396..5984
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5885..5904
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6138..6155
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact
61 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
121 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
181 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
241 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
301 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
361 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
421 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
481 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
541 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
601 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
661 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
721 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
781 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
841 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
901 gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
961 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
1021 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
1081 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
1141 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
1201 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
1261 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
1321 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
1381 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
1441 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta
1501 tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg
1561 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
1621 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
1681 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
1741 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
1801 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
1861 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
1921 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
1981 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
2041 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
2101 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
2161 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
2221 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
2281 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
2341 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg
2401 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag
2461 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa
2521 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
2581 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
2641 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat
2701 cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta
2761 gaaataattt tgtttaactt taagaaggag atatacatat gaaatacctg ctgccgaccg
2821 ctgctgctgg tctgctgctc ctcgctgccc agccggccat ggccgaggtg cagctggtgc
2881 agtctggggc tgaggtgaag aagcctgggg cctcagtgaa ggtctcctgc aaggcttctg
2941 gttacacctt taccagctat agtatcagct gggtgcgaca ggcccctgga caagggcttg
3001 agtggatggg atggatcagc gcttacaatg gtaacacaag ctatgcacag aagctccagg
3061 gcagagtcac gattaccgcg gacaaatcca cgagcacagc ctacatggag ctgagcagcc
3121 tgagatctga ggacacggcc gtgtattact gtgcgaaggc cgggcaccgt tcagggagat
3181 ataacaactg gttcgacccc tggggccagg gcaccctggt caccgtctcc tcactcgagg
3241 gtggaggcgg ttcaggcgga ggtggctctg gcggtggcgc tagccagtct gagctgactc
3301 agcctccctc ggtgtcagtg gccccaggac agacggccag gatcacctgt ggggaaaaca
3361 acattggaag taaaaatgtg cactggtacc agcagaagcc aggccaggcc cctgtgctga
3421 tcatctatag gggtagcaac cggccctctg ggatccctga gcgattctct ggctccaact
3481 cggggaacac ggccaccctg accatcagca gagcccaagc cggggatgag gctgactatt
3541 actgccaggt gtttgatagg aggagtgacc atccggtggt gttcggcgga gggaccaagc
3601 tgaccgtcct aggtgcggcc gcatccgcac atcatcatca ccatcacaag ctggactaca
3661 aagaccatga cggtgattat aaagatcatg acatcgatta caaggatgac gatgacaagt
3721 aataaaagct ttaataagtc gagcaccacc accaccacca ctgagatccg gctgctaaca
3781 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
3841 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat
3901 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
3961 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
4021 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
4081 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
4141 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
4201 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
4261 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
4321 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
4381 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
4441 ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
4501 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
4561 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
4621 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
4681 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
4741 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
4801 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
4861 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
4921 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
4981 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
5041 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
5101 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
5161 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
5221 gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcataacacc
5281 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgaccaaa atcccttaac
5341 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
5401 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
5461 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
5521 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
5581 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
5641 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
5701 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
5761 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
5821 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
5881 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
5941 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
6001 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
6061 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
6121 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcg
//
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