| 质粒名称: | pHH0103_SASH1-1/1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 91347 |
| 存储实验室: | Sachdev Sidhu |
|
载体骨架基本信息 |
|
| 载体名称: | pHH0103 |
| 载体出品公司: | -- |
| 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: | 细菌表达 |
| 筛选标记: | -- |
| 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: | SASH1-1/1 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 227 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | Sfi1（销毁后未知） |
| 3' 克隆位点: | Not1（销毁后未知） |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pHH0103_SASH1-1/1 was a gift from Sachdev Sidhu (Addgene plasmid # 91347 ; http://n2t.net/addgene:91347 ; RRID:Addgene_91347) |
| 参考文献部分: | Comprehensive Analysis of the Human SH3 Domain Family Reveals a Wide Variety of Non-canonical Specificities. Teyra J, Huang H, Jain S, Guan X, Dong A, Liu Y, Tempel W, Min J, Tong Y, Kim PM, Bader GD, Sidhu SS. Structure. 2017 Oct 3;25(10):1598-1610.e3. doi: 10.1016/j.str.2017.07.017. Epub 2017 Sep 7. 10.1016/j.str.2017.07.017 PubMed 28890361 |
买家导航
pHH0103_SASH1-1/1载体质粒基本信息
pHH0103_SASH1-1/1质粒图谱载体图谱和pHH0103_SASH1-1/1载体序列质粒序列多克隆位点信息
pHH0103_SASH1-1/1质粒载体简介
pHH0103_SASH1-1/1质粒序列载体序列
LOCUS pHH0103_SASH1-1/1 6918 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Protein expression and purification of human SH3 domain construct
SASH1-1/1.
ACCESSION .
VERSION .
KEYWORDS pHH0103_SASH1-1/1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6918)
AUTHORS Teyra J, Huang H, Jain S, Guan X, Dong A, Liu Y, Tempel W, Min J,
Tong Y, Kim PM, Bader GD, Sidhu SS
TITLE Comprehensive Analysis of the Human SH3 Domain Family Reveals a Wide
Variety of Non-canonical Specificities.
JOURNAL Structure. 2017 Oct 3;25(10):1598-1610.e3. doi:
10.1016/j.str.2017.07.017. Epub 2017 Sep 7.
PUBMED 28890361
REFERENCE 2 (bases 1 to 6918)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6918
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 5..23
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 46..123
/gene="lacI (mutant)"
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
CDS 124..1206
/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 complement(143..162)
/label=LacI-R
/note="LacI, reverse primer"
promoter 1449..1477
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 1485..1501
/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)."
primer_bind 1490..1512
/label=M13/pUC Reverse
/note="In lacZ gene"
CDS 1586..1603
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 1619..2272
/codon_start=1
/product="glutathione S-transferase from Schistosoma
japonicum"
/label=GST
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
primer_bind 2230..2252
/label=pGEX 5'
/note="pGEX vectors, Glutathione-S-transferase, forward
primer"
CDS 2279..2302
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
CDS 2315..2332
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
terminator 2561..2608
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
rep_origin complement(4201..4656)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4338..4359)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4550..4569
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind complement(4698..4720)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 4806..4946
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 4861..4880
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(4961..4978)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5132..5719)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5890..6750)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 6513..6532
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6751..6855)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 taaaaatagg cgtatcacga ggccctttcg tcttcaagaa ttcccgacac catcgaatgg
61 tgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg
121 aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc
181 gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa
241 gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa
301 cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt
361 gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta
421 gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc
481 agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc
541 tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt
601 attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac
661 cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct
721 ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac
781 tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc
841 actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag
901 tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc
961 tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc
1021 agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg
1081 cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc
1141 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
1201 cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcacaat tctcatgttt
1261 gacagcttat catcgactgc acggtgcacc aatgcttctg gcgtcaggca gccatcggaa
1321 gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct caaggcgcac
1381 tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa atattctgaa
1441 atgagctgtt gacaattaat catcggctcg tataatgtgt ggaattgtga gcggataaca
1501 atttcacaca ggaaacagcc agtccgttta ggtgttttca cgagcacttc accaacaagg
1561 accatagatt atgaaaatcg aagaacacca tcaccatcac cattccagcg gtaagcttat
1621 gtcccctata ctaggttatt ggaaaattaa gggccttgtg caacccactc gacttctttt
1681 ggaatatctt gaagaaaaat atgaagagca tttgtatgag cgcgatgaag gtgataaatg
1741 gcgaaacaaa aagtttgaat tgggtttgga gtttcccaat cttccttatt atattgatgg
1801 tgatgttaaa ttaacacagt ctatggccat catacgttat atagctgaca agcacaacat
1861 gttgggtggt tgtccaaaag agcgtgcaga gatttcaatg cttgaaggag cggttttgga
1921 tattagatac ggtgtttcga gaattgcata tagtaaagac tttgaaactc tcaaagttga
1981 ttttcttagc aagctacctg aaatgctgaa aatgttcgaa gatcgtttat gtcataaaac
2041 atatttaaat ggtgatcatg taacccatcc tgacttcatg ttgtatgacg ctcttgatgt
2101 tgttttatac atggacccaa tgtgcctgga tgcgttccca aaattagttt gttttaaaaa
2161 acgtattgaa gctatcccac aaattgataa gtacttgaaa tccagcaagt atatagcatg
2221 gcctttgcag ggctggcaag ccacgtttgg tggtggcgac catcctccaa aatcggatct
2281 ggaagttctg ttccaggggc ccctgtccag cggtctggtt ccgcgtggtt ccggtaccgc
2341 ggcccagccg gccatggccc agggcgcgct gggtcgtgct cgtgttcata ccgatttcac
2401 cccatctccg tatgataccg attccctgaa actgaaaaaa ggtgacatta tcgacatcat
2461 ttccaaacca ccgatgggta cttggatggg tctgctgaac aacaaagttg gtactttcaa
2521 attcatttac gttgatgttc tgtccgcggc cgcataataa accgatacaa ttaaaggctc
2581 cttttggagc cttttttttg gagattttca acgtgaaaaa attattattc gcaattcctt
2641 tagttgttcc tttctattct cactccgctg aaactgttga aagttgttta gcaaaacccc
2701 atacagaaaa ttcatttact aacgtctgga aagacgacaa aactttagat cgttacgcta
2761 actatgaggg ctgtctgtgg aatgctacag gcgttgtagt ttgtactggt gacgaaactc
2821 agtgtctagc tagagtggcg gtggctctgg ttccggtgat tttgattatg aaaagatggc
2881 aaacgctaat aagggggcta tgaccgaaaa tgccgatgaa aacgcgctac agtctgacgc
2941 taaaggcaaa cttgattctg tcgctactga ttacggtgct gctatcgatg gtttcattgg
3001 tgacgtttcc ggccttgcta atggtaatgg tgctactggt gattttgctg gctctaattc
3061 ccaaatggct caagtcggtg acggtgataa ttcaccttta atgaataatt tccgtcaata
3121 tttaccttcc ctccctcaat cggttgaatg tcgccctttt gtctttggcg ctggtaaacc
3181 atatgaattt tctattgatt gtgacaaaat aaacttattc cgtggtgtct ttgcgtttct
3241 tttatatgtt gccaccttta tgtatgtatt ttctacgttt gctaacatac tgcgtaataa
3301 ggagtcttaa tcatgccagt tcttttggct agcgccgccc tataccttgt ctgcctcccc
3361 gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct gaatggaagc cggcggcacc
3421 tcgctaacgg attcaccact ccaagaattg gagccaatca attcttgcgg agaactgtga
3481 atgcgcaaac caacccttgg cagaacatat ccatcgcgtc cgccatctcc agcagccgca
3541 cgcggcgcat ctcgggcagc gttgggtcct ggccacgggt gcgcatgatc gtgctcctgt
3601 cgttgaggac ccggctaggc tggcggggtt gccttactgg ttagcagaat gaatcaccga
3661 tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga gcaacaacat
3721 gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca
3781 ccattatgtt ccggatctgc atcgcaggat gctgctggct accctgtgga acacctacat
3841 ctgtattaac gaagcgctgg cattgaccct gagtgatttt tctctggtcc cgccgcatcc
3901 ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa
3961 cccgtatcgt gagcatcctc tctcgtttca tcggtatcat tacccccatg aacagaaatc
4021 ccccttacac ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac atggcccgct
4081 ttatcagaag ccagacatta acgcttctgg agaaactcaa cgagctggac gcggatgaac
4141 aggcagacat ctgtgaatcg cttcacgacc acgctgatga gctttaccgc aggatccgcg
4201 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
4261 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
4321 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
4381 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
4441 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
4501 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
4561 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
4621 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcgcg gatcctgcct cgcgcgtttc
4681 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
4741 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
4801 cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg
4861 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
4921 gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc
4981 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
5041 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
5101 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
5161 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
5221 agggtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
5281 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
5341 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
5401 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
5461 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
5521 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
5581 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
5641 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
5701 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
5761 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
5821 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
5881 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
5941 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
6001 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
6061 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
6121 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
6181 tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat
6241 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatattgtg
6301 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
6361 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
6421 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
6481 accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt
6541 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
6601 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
6661 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
6721 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
6781 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
6841 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
6901 tatcatgaca ttaaccta
//
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