| 质粒名称: | AviTag-SpyCatcher |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 72326 |
| 存储实验室: | Mark Howarth |
|
载体骨架基本信息 |
|
| 载体名称: | pDEST14-SpyCatcher |
| 载体出品公司: | Howarth lab (Addgene plasmid #35044) |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6422 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: | AviTag-SpyCatcher |
| 物种来源: | Synthetic |
| 插入大小: | 474 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | N-末端His6标记和TEV裂解位点 |
| 克隆方法: | Ligation Independent 克隆 |
| 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: | AviTag-SpyCatcher was a gift from Mark Howarth (Addgene plasmid # 72326 ; http://n2t.net/addgene:72326 ; RRID:Addgene_72326) |
| 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
AviTagSpyCatcher载体基本信息
AviTagSpyCatcher载体图谱质粒图谱和多克隆位点信息
AviTagSpyCatcher载体简介
AviTagSpyCatcher载体序列
LOCUS AviTag-SpyCatcher 4115 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Bacterial expression of SpyCatcher with a peptide tag enabling
site-specific biotinylation..
ACCESSION .
VERSION .
KEYWORDS AviTag-SpyCatcher
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4115)
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 4115)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4115
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 21..40
/label=T7
/note="T7 promoter, forward primer"
promoter 21..39
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 71..93
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 113..130
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 137..181
/codon_start=1
/product="peptide tag that allows for enzymatic
biotinylation"
/label=AviTag(TM)
/translation="GLNDIFEAQKIEWHE"
CDS 206..226
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
CDS 636..653
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(706..724)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 720..767
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 804..1259
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(891..910)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1101..1122
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 1285..1389
/gene="bla"
/label=AmpR promoter
CDS 1390..2250
/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 complement(1608..1627)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2421..3009
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2910..2929
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3163..3180
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 3195..3334
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(3260..3279)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3420..3442
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(3436..3627)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
ORIGIN
1 agatctcgat cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc
61 tagaaataat tttgtttaac tttaagaagg agatatacat atgtcgtact accatcacca
121 tcaccatcac ggcagcggcc tgaacgatat ttttgaagcg cagaaaattg aatggcatga
181 aggcgattac gacatcccaa cgaccgaaaa cctgtatttt cagggcgcca tggttgatac
241 cttatcaggt ttatcaagtg agcaaggtca gtccggtgat atgacaattg aagaagatag
301 tgctacccat attaaattct caaaacgtga tgaggacggc aaagagttag ctggtgcaac
361 tatggagttg cgtgattcat ctggtaaaac tattagtaca tggatttcag atggacaagt
421 gaaagatttc tacctgtatc caggaaaata tacatttgtc gaaaccgcag caccagacgg
481 ttatgaggta gcaactgcta ttacctttac agttaatgag caaggtcagg ttactgtaaa
541 tggcaaagca actaaaggtg acgctcatat ttaaatggtt gatgcttgag gatccgaatt
601 cgagctccgt cgacaagctt gcgaccgcac tcgagcacca ccaccaccac cactgagatc
661 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
721 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa
781 ctatatccgg attggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
841 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
901 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
961 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
1021 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
1081 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
1141 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
1201 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
1261 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
1321 tcaaatatgt atccgctcat gagacaataa ccctggtaaa tgcttcaata atattgaaaa
1381 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
1441 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
1501 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
1561 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
1621 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
1681 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
1741 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
1801 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
1861 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
1921 accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
1981 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
2041 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
2101 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
2161 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
2221 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
2281 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
2341 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
2401 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
2461 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
2521 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
2581 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
2641 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
2701 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
2761 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
2821 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
2881 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
2941 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
3001 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
3061 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
3121 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
3181 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
3241 cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
3301 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
3361 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
3421 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
3481 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
3541 cagctcgttg agcttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
3601 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
3661 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
3721 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
3781 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc
3841 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga
3901 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca
3961 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt
4021 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat
4081 catgcgcacc cgtggccagg acccaacgct gcccg
//
