| 质粒名称: | pDJSVT86 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 108965 |
| 存储实验室: | Daniel Slade |
|
载体骨架基本信息 |
|
| 载体名称: | pET-16b |
| 载体出品公司: | EMD Millipore |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5711 bp |
| 完整质粒大小: | -- |
| 载体修饰: | 移除NcoI和XhoI位点之间的DNA，并替换为新序列。 |
| 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: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | T7 |
| 标签/融合蛋白: | OmpA 1-27，6His（主干上的N终端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | T7 promoter |
| 3' 测序引物 | T7 terminator |
|
文献引用方法 |
|
| 材料和方法部分: | pDJSVT86 was a gift from Daniel Slade (Addgene plasmid # 108965 ; http://n2t.net/addgene:108965 ; RRID:Addgene_108965) |
| 参考文献部分: | A chemical and biological toolbox for Type Vd secretion: Characterization of the phospholipase A1 autotransporter FplA from Fusobacterium nucleatum. Casasanta MA, Yoo CC, Smith HB, Duncan AJ, Cochrane K, Varano AC, Allen-Vercoe E, Slade DJ. J Biol Chem. 2017 Dec 8;292(49):20240-20254. doi: 10.1074/jbc.M117.819144. Epub 2017 Oct 11. 10.1074/jbc.M117.819144 PubMed 29021252 |
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 pDJSVT86 5762 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Express periplasmic, outer membrane, or secreted proteins in E. coli
- OmpA signal and 6His tag.
ACCESSION .
VERSION .
KEYWORDS pDJSVT86
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5762)
AUTHORS Casasanta MA, Yoo CC, Smith HB, Duncan AJ, Cochrane K, Varano AC,
Allen-Vercoe E, Slade DJ
TITLE A chemical and biological toolbox for Type Vd secretion:
Characterization of the phospholipase A1 autotransporter FplA from
Fusobacterium nucleatum.
JOURNAL J Biol Chem. 2017 Dec 8;292(49):20240-20254. doi:
10.1074/jbc.M117.819144. Epub 2017 Oct 11.
PUBMED 29021252
REFERENCE 2 (bases 1 to 5762)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5762
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 10..38
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
terminator 213..260
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 256..274
/label=T7 Term
/note="T7 terminator, reverse primer"
CDS complement(345..362)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
sig_peptide complement(381..443)
/product="signal peptide from the E. coli outer membrane
protein OmpA"
/label=OmpA signal peptide
RBS 456..478
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 493..517
/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(517..536)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(518..536)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(620..639)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter 845..922
/gene="lacI"
/label=lacI promoter
CDS 923..2005
/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(942..961)
/label=LacI-R
/note="LacI, reverse primer"
CDS 3317..3508
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
primer_bind complement(3502..3524)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 3610..3749
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 3665..3684
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(3764..3781)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3935..4523)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4015..4034)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4694..5554)
/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 5317..5336
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5555..5659)
/gene="bla"
/label=AmpR promoter
primer_bind 5727..5745
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
181 gcgggatatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag
241 gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt
301 cgggctttgt tagcagccgg atcctcgagg gtacccgcgg ccgcgtggtg gtggtggtgg
361 tgggtgttat ctttcggagc ggcctgcgct acggtagcga aaccagccag tgccactgca
421 atcgcgatag ctgtcttttt cattcccatg gtatatctcc ttcttaaagt taaacaaaat
481 tatttctaga ggggaattgt tatccgctca caattcccct atagtgagtc gtattaattt
541 cgcgggatcg agatctcgat cctctacgcc ggacgcatcg tggccggcat caccggcgcc
601 acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc
661 cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg
721 ggactgttgg gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc
781 ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgagat
841 cccggacacc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga
901 gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc
961 cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa
1021 aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc
1081 acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct
1141 gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag
1201 cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa
1261 tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc
1321 cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca
1381 gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca
1441 tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc
1501 ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat
1561 agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct
1621 gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc
1681 aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata
1741 cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca aacaggattt
1801 tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt
1861 gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa
1921 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
1981 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt
2041 aggcaccggg atctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt
2101 gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg
2161 taggacaggt gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg
2221 cgacgatgat cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct
2281 tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg
2341 cggccgacgc gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc
2401 ccattatgat tcttctcgct tccggcggca tcgggatgcc cgcgttgcag gccatgctgt
2461 ccaggcaggt agatgacgac catcagggac agcttcaagg atcgctcgcg gctcttacca
2521 gcctaacttc gatcattgga ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca
2581 catggaacgg gttggcatgg attgtaggcg ccgccctata ccttgtctgc ctccccgcgt
2641 tgcgtcgcgg tgcatggagc cgggccacct cgacctgaat ggaagccggc ggcacctcgc
2701 taacggattc accactccaa gaattggagc caatcaattc ttgcggagaa ctgtgaatgc
2761 gcaaaccaac ccttggcaga acatatccat cgcgtccgcc atctccagca gccgcacgcg
2821 gcgcatctcg ggcagcgttg ggtcctggcc acgggtgcgc atgatcgtgc tcctgtcgtt
2881 gaggacccgg ctaggctggc ggggttgcct tactggttag cagaatgaat caccgatacg
2941 cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa caacatgaat
3001 ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc cctgcaccat
3061 tatgttccgg atctgcatcg caggatgctg ctggctaccc tgtggaacac ctacatctgt
3121 attaacgaag cgctggcatt gaccctgagt gatttttctc tggtcccgcc gcatccatac
3181 cgccagttgt ttaccctcac aacgttccag taaccgggca tgttcatcat cagtaacccg
3241 tatcgtgagc atcctctctc gtttcatcgg tatcattacc cccatgaaca gaaatccccc
3301 ttacacggag gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg cccgctttat
3361 cagaagccag acattaacgc ttctggagaa actcaacgag ctggacgcgg atgaacaggc
3421 agacatctgt gaatcgcttc acgaccacgc tgatgagctt taccgcagct gcctcgcgcg
3481 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
3541 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
3601 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac
3661 tatgcggcat cagagcagat tgtactgaga gtgcaccatt gcggtgtgaa ataccgcaca
3721 gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc
3781 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
3841 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
3901 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
3961 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
4021 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
4081 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
4141 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
4201 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
4261 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
4321 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
4381 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
4441 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
4501 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
4561 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
4621 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
4681 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
4741 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
4801 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
4861 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
4921 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
4981 atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg
5041 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
5101 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
5161 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
5221 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
5281 ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa
5341 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
5401 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
5461 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
5521 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
5581 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
5641 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
5701 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcaag
5761 aa
//
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