| 质粒名称: | pET28a SpyCatcher-SnoopCatcher |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 72324 |
| 存储实验室: | Mark Howarth |
|
载体骨架基本信息 |
|
| 载体名称: | pET21b |
| 载体出品公司: | Novagen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5442 bp |
| 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: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | pET28a SpyCatcher-SnoopCatcher,SpyCatcher-SnoopCatcher |
| 物种来源: | Synthetic |
| 插入大小: | 774 bp |
| 基因突变: | 删除了SpyCatcher N端的35个氨基酸 |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | N端子His6标签 |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | TAA TAC GAC TCA CTA TAG GG |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pET28a SpyCatcher-SnoopCatcher was a gift from Mark Howarth (Addgene plasmid # 72324 ; http://n2t.net/addgene:72324 ; RRID:Addgene_72324) |
| 参考文献部分: | Programmable polyproteams built using twin peptide superglues. Veggiani G, Nakamura T, Brenner MD, Gayet RV, Yan J, Robinson CV, Howarth M. Proc Natl Acad Sci U S A. 2016 Jan 19. pii: 201519214. 10.1073/pnas.1519214113 PubMed 26787909 |
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 pET28a SpyCatcher-SnoopCatcher 6136 bp ds-DNA circular SYN 21-AUG-2019
DEFINITION Bacterial expression of SpyCatcher linked to SnoopCatcher..
ACCESSION .
VERSION .
KEYWORDS pET28a SpyCatcher-SnoopCatcher
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6136)
AUTHORS Veggiani G, Nakamura T, Brenner MD, Gayet RV, Yan J, Robinson CV,
Howarth M
TITLE Programmable polyproteams built using twin peptide superglues.
JOURNAL Proc Natl Acad Sci U S A. 2016 Jan 19. pii: 201519214.
PUBMED 26787909
REFERENCE 2 (bases 1 to 6136)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 21, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6136
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin complement(1..456)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(138..159)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 350..369
/label=F1ori-R
/note="F1 origin, reverse primer"
terminator 493..540
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 536..554
/label=T7 Term
/note="T7 terminator, reverse primer"
CDS complement(607..624)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS complement(1375..1392)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
RBS 1412..1434
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 1449..1473
/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 complement(1473..1492)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1474..1492)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1576..1595)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter 1801..1878
/gene="lacI"
/label=lacI promoter
CDS 1879..2961
/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(1898..1917)
/label=LacI-R
/note="LacI, reverse primer"
CDS 3770..3961
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
primer_bind complement(3955..3977)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 4063..4202
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 4118..4137
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(4217..4234)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4388..4975)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4468..4487)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5146..6006)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 5769..5788
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6007..6111)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
181 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccaatc cggatatagt
481 tcctcctttc agcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctag
541 ttattgctca gcggtggcag cagccaactc agcttccttt cgggctttgt tagcagccgg
601 atctcagtgg tggtggtggt ggtgctcgag ttattatttc ggcggtatcg gttcattggt
661 gatataatgt ttaccgttgg taaattcata tgtagccgga atatcctgcg gcacaatgct
721 ggtcacatca cgcacttcgc cattcacaat ctgaaacgcc acaatcggct tattctgcac
781 cggtttatag ccagcgggtt cgctattttc aaacaggcga tatttgccat cgctcagatt
841 cttaaaggtc agtttaccat cttcgccggt acgcacattt tgataggtcc cattctgatc
901 aatcgcgcca tagatatcgg gatagtcggg atgctgtttc tgcaggctaa acacggcacc
961 acgcagcggc ttcatatggg atcccttctc cagctgctcc ttcagcttct tggcgttggc
1021 cagctcctcg gcggcctgtc tctgctcctt ctgcttctgg gcctccaggg ccttcaggtt
1081 ggcgggggat ccaatatgag cgtcaccttt agttgctttg ccatttacag taacctgacc
1141 ttgctcatta actgtaaagg taatagcagt tgctacctca taaccgtctg gtgctgcggt
1201 ttcgacaaat gtatattttc ctggatacag gtagaaatct ttcacttgtc catctgaaat
1261 ccatgtacta atagttttac cagatgaatc acgcaactcc atagttgcac cagctaactc
1321 tttgccgtcc tcatcacgtt ttgagaattt aatatgggta gcactatcgt aatcgtgatg
1381 gtgatggtga tggtagtacg acatatgtat atctccttct taaagttaaa caaaattatt
1441 tctagagggg aattgttatc cgctcacaat tcccctatag tgagtcgtat taatttcgcg
1501 ggatcgagat ctcgatcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag
1561 gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact
1621 tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac
1681 tgttgggcgc catctccttg catgcaccat tccttgcggc ggcggtgctc aacggcctca
1741 acctactact gggctgcttc ctaatgcagg agtcgcataa gggagagcgt cgagatcccg
1801 gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt
1861 caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt
1921 gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg
1981 cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa
2041 caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac
2101 gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg
2161 gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt
2221 ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt
2281 gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca
2341 cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg
2401 gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg
2461 cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg
2521 gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat
2581 gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg
2641 cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac
2701 gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc
2761 ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag
2821 ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg
2881 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
2941 cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc
3001 accgggatct cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc
3061 gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg
3121 acaggtgccg gcagcgctct gggtcatttt cggcgaggac cgctttcgct ggagcgcgac
3181 gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc aagccttcgt
3241 cactggtccc gccaccaaac gtttcggcga gaagcaggcc attatcgccg gcatggcggc
3301 cccacgggtg cgcatgatcg tgctcctgtc gttgaggacc cggctaggct ggcggggttg
3361 ccttactggt tagcagaatg aatcaccgat acgcgagcga acgtgaagcg actgctgctg
3421 caaaacgtct gcgacctgag caacaacatg aatggtcttc ggtttccgtg tttcgtaaag
3481 tctggaaacg cggaagtcag cgccctgcac cattatgttc cggatctgca tcgcaggatg
3541 ctgctggcta ccctgtggaa cacctacatc tgtattaacg aagcgctggc attgaccctg
3601 tgtgattttt ctctggtccc gccgcatcca taccgccagt tgtttaccct cacaacgttc
3661 cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg agcatcctct ctcgtttcat
3721 cggtatcatt acccccatga acagaaatcc cccttacacg gaggcatcag tgaccaaaca
3781 ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc cagacattaa cgcttctgga
3841 gaaactcaac gagctggacg cggatgaaca ggcagacatc tgtgaatcgc ttcacgacca
3901 cgctgatgag ctttaccgca gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg
3961 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
4021 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc
4081 acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg
4141 agagtgcacc attgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
4201 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
4261 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
4321 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
4381 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
4441 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
4501 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
4561 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
4621 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
4681 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
4741 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
4801 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
4861 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
4921 gcggtggttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
4981 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
5041 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
5101 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
5161 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
5221 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
5281 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
5341 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
5401 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
5461 tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
5521 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
5581 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
5641 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
5701 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
5761 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
5821 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
5881 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
5941 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
6001 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
6061 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
6121 ccgaaaagtg ccacct
//
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