| 质粒名称: | pET21a(+)-Bst LF-6xHis |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 159148 |
| 存储实验室: | Andrea Pauli |
|
载体骨架基本信息 |
|
| 载体名称: | pET21a |
| 载体出品公司: | EMD Bioscience |
| 空载体大小: | 5443 bp |
| 完整质粒大小: | 7137 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | large fragment of Geobacillus stearothermophilus DNA polymerase I,Bst LF |
| 物种来源: | Geobacillus stearothermophilus |
| 插入大小: | 1776 bp |
| 基因突变: | -- |
| GenBank编号 | AAB52611.1 |
| 启动子: | -- |
| 标签/融合蛋白: | His（插入件上的C端子） |
| 克隆方法: | -- |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pET21a(+)-Bst LF-6xHis was a gift from Andrea Pauli (Addgene plasmid # 159148 ; http://n2t.net/addgene:159148 ; RRID:Addgene_159148) |
| 参考文献部分: | A rapid, highly sensitive and open-access SARS-CoV-2 detection assay for laboratory and home testing. Kellner MJ, Ross JJ, Schnabl J, Dekens MPS, Heinen R, Grishkovskaya I, Bauer B, Stadlmann J, Menéndez-Arias L, Fritsche-Polanz R, Traugott M, Seitz T, Zoufaly A, Födinger M, Wenisch C, Zuber J, Pauli A, Brennecke J.. bioRxiv. 2020. 10.1101/2020.06.23.166397 |
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
-Bst LF-6xHis质粒图谱.png)
载体简介
载体序列
LOCUS pET21a(+)-Bst LF-6xHis 7133 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION expresses His-tagged large fragment of Bst polymerase.
ACCESSION .
VERSION .
KEYWORDS pET21a(+)-Bst LF-6xHis
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7133)
AUTHORS Kellner MJ, Ross JJ, Schnabl J, Dekens MPS, Heinen R, Grishkovskaya
I, Bauer B, Stadlmann J, Menendez-Arias L, Fritsche-Polanz R,
Traugott M, Seitz T, Zoufaly A, Fodinger M, Wenisch C, Zuber J,
Pauli A, Brennecke J.
TITLE A rapid, highly sensitive and open-access SARS-CoV-2 detection assay
for laboratory and home testing.
JOURNAL bioRxiv. 2020.
REFERENCE 2 (bases 1 to 7133)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7133
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 563..582
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(602..679)
/gene="lacI"
/label=lacI promoter
primer_bind 885..904
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 988..1007
/label=T7
/note="T7 promoter, forward primer"
promoter 988..1006
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 1007..1031
/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 1046..1068
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 2852..2869
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(2922..2940)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 2936..2983
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 3020..3475
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3107..3126)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3317..3338
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 3501..3605
/gene="bla"
/label=AmpR promoter
CDS 3606..4466
/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(3824..3843)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4637..5225
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5126..5145
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5379..5396
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 5411..5550
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(5476..5495)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5636..5658
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(5652..5843)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(6652..601)
/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"
ORIGIN
1 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
61 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
121 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
181 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
241 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
301 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
361 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
421 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
481 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
541 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
601 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
661 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
721 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
781 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
841 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
901 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
961 aggatcgaga tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg
1021 ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata tacatatggc
1081 ggagggagaa aagccgttgg aagaaatgga gtttgctatt gttgatgtaa ttaccgaaga
1141 gatgctggca gataaagccg ctttagtggt tgaagtaatg gaggaaaatt accacgacgc
1201 ccctatcgtt ggaattgcgt tggtcaatga acatgggcgc tttttcatgc gccctgagac
1261 tgctcttgct gactcacaat ttcttgcttg gctggccgat gagactaaga agaaatctat
1321 gttcgatgca aagcgtgccg ttgttgcttt gaagtggaaa ggcatcgaac ttcgcggtgt
1381 agctttcgac ctgctgcttg cagcatacct tcttaatcca gctcaggatg cgggcgacat
1441 cgcggctgta gcgaaaatga aacaatatga agctgttcgt tccgatgagg cagtctacgg
1501 caagggagtc aaacgttctc ttccggatga acagacatta gccgagcatc tggtccgtaa
1561 ggccgcagcc atctgggcac tggagcaacc tttcatggat gacctgcgca acaatgaaca
1621 ggaccagctt ctgacaaagc tggaacagcc gttggcggca attttggcgg aaatggaatt
1681 tactggcgtc aatgtggaca cgaagcgctt agaacaaatg ggatctgaac ttgccgagca
1741 gttacgtgca atcgagcaac gtatttatga gctggcggga caggagttca atatcaatag
1801 cccaaaacag ttgggggtaa tcctgtttga aaaactgcag ctgcccgtat tgaagaaaac
1861 gaagaccggt tattcgacaa gcgctgatgt gttggaaaaa ttggcacccc accacgagat
1921 cgtagagaat atcttgcatt accgccagtt gggcaagttg caaagcacgt atatcgaagg
1981 ccttcttaag gtggtacgcc cagacacggg gaaagtccat acgatgttta accaagcttt
2041 aacccaaact ggtcgcctta gtagtgcaga acccaattta caaaatattc ccatccgttt
2101 agaagaaggc cgtaaaattc gtcaggcctt tgtgccatct gaacccgatt ggttgatttt
2161 tgcagcggat tatagccaaa ttgagttacg cgtgttggca catatcgctg acgacgataa
2221 tctgatcgaa gcatttcagc gcgatttgga catccatact aagacagcga tggacatctt
2281 ccatgtaagt gaggaggaag tgacggcgaa catgcgccgc caggctaaag ctgttaactt
2341 cggtatcgta tacggaatca gcgactacgg gttggctcag aacttgaaca ttacgcgtaa
2401 ggaagcagcg gagtttattg agcgctactt cgcctctttc ccaggtgtta agcaatatat
2461 ggaaaacatc gttcaggagg cgaaacaaaa ggggtatgtc acgacacttc ttcatcgccg
2521 ccgttacctg cccgatatca ccagtcgtaa ctttaacgtg cgtagtttcg cggagcgcac
2581 ggcgatgaat acccccattc agggttccgc agccgacatt atcaaaaagg cgatgattga
2641 cttggccgcg cgcttaaagg aagagcaact tcaagcgcgc ctgttactgc aggtacatga
2701 tgagcttatc ttagaggctc ccaaagagga gatcgaacgt ttgtgtgagt tggttcccga
2761 agtcatggag caagctgtta cgttacgcgt accattaaag gtagactacc actacggtcc
2821 tacatggtat gacgcgaaag gttcttccgg gcaccaccac caccaccact gagatccggc
2881 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
2941 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
3001 atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
3061 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
3121 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
3181 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
3241 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
3301 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
3361 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
3421 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
3481 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
3541 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
3601 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
3661 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
3721 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
3781 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
3841 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
3901 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
3961 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
4021 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
4081 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
4141 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
4201 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
4261 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
4321 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
4381 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
4441 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
4501 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
4561 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
4621 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
4681 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
4741 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
4801 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
4861 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
4921 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
4981 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
5041 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
5101 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
5161 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
5221 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
5281 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
5341 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
5401 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
5461 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
5521 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
5581 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
5641 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
5701 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
5761 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
5821 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
5881 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
5941 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
6001 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
6061 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
6121 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
6181 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
6241 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
6301 cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg
6361 gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac
6421 aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct
6481 gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata
6541 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
6601 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
6661 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
6721 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
6781 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
6841 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
6901 gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
6961 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
7021 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
7081 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gcc
//
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