| 质粒名称: | pET21b-Casp1-His |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 11809 |
| 存储实验室: | Guy Salvesen |
|
载体骨架基本信息 |
|
| 载体名称: | pET-21 b |
| 载体出品公司: | Clontech |
| 空载体大小: | 5443 bp |
| 完整质粒大小: | 6575 bp |
| 载体类型: | 细菌细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 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: | Caspase-1 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1350 bp |
| 标签/融合蛋白: | His (C terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | XhoI(未破坏) |
| 3’克隆位点: | NdeI (未破坏) |
| 5’测序引物 | T7 |
|
文献引用方法 |
|
| 材料和方法部分: | pET21b-Casp1-His was a gift from Guy Salvesen (Addgene plasmid # 11809 ; http://n2t.net/addgene:11809 ; RRID:Addgene_11809) |
买家导航
pET21b-Casp1-His载体质粒基本信息
pET21b-Casp1-His质粒图谱载体图谱和pET21b-Casp1-His载体序列质粒序列多克隆位点信息
pET21b-Casp1-His质粒载体简介
pET21b-Casp1-His质粒序列载体序列
LOCUS pET21b-Casp1-His 6575 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pET21b-Casp1-His
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6575)
TITLE Salvesen Casp1, Casp2 plasmids
REFERENCE 2 (bases 1 to 6575)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6575
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 171..190
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 274..293
/label=T7
/note="T7 promoter, forward primer"
promoter 274..292
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 293..317
/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 332..354
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 1580..1597
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(1650..1668)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 1664..1711
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 1748..2203
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1835..1854)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2045..2066
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 2229..2333
/gene="bla"
/label=AmpR promoter
CDS 2334..3194
/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(2552..2571)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3365..3953
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3854..3873
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4107..4124
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 4139..4278
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(4204..4223)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4364..4386
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(4380..4571)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(5380..6462)
/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 6424..6443
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(6463..6540)
/gene="lacI"
/label=lacI promoter
ORIGIN
1 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt
61 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
121 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
181 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
241 gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt
301 gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatataca
361 tatggccgac aaggtcctga aggagaagag aaagctgttt atccgttcca tgggtgaagg
421 tacaataaat ggcttactgg atgaattatt acagacaagg gtgctgaaca aggaagagat
481 ggagaaagta aaacgtgaaa atgctacagt tatggataag acccgagctt tgattgactc
541 cgttattccg aaaggggcac aggcatgcca aatttgcatc acatacattt gtgaagaaga
601 cagttacctg gcagggacgc tgggactctc agcagatcaa acatctggaa attaccttaa
661 tatgcaagac tctcaaggag tactttcttc ctttccagct cctcaggcag tgcaggacaa
721 cccagctatg cccacatcct caggctcaga agggaatgtc aagctttgct ccctagaaga
781 agctcaaagg atatggaaac aaaagtcggc agagatttat ccaataatgg acaagtcaag
841 ccgcacacgt cttgctctca ttatctgcaa tgaagaattt gacagtattc ctagaagaac
901 tggagctgag gttgacatca caggcatgac aatgctgcta caaaatctgg ggtacagcgt
961 agatgtgaaa aaaaatctca ctgcttcgga catgactaca gagctggagg catttgcaca
1021 ccgcccagag cacaagacct ctgacagcac gttcctggtg ttcatgtctc atggtattcg
1081 ggaaggcatt tgtgggaaga aacactctga gcaagtccca gatatactac aactcaatgc
1141 aatctttaac atgttgaata ccaagaactg cccaagtttg aaggacaaac cgaaggtgat
1201 catcatccag gcctgccgtg gtgacagccc tggtgtggtg tggtttaaag attcagtagg
1261 agtttctgga aacctatctt taccaactac agaagagttt gaggatgatg ctattaagaa
1321 agcccacata gagaaggatt ttatcgcttt ctgctcttcc acaccagata atgtttcttg
1381 gagacatccc acaatgggct ctgtttttat tggaagactc attgaacaca tgcaagaata
1441 tgcctgttcc tgtgatgtgg aggaaatttt ccgcaaggtt cgattttcat ttgagcagcc
1501 agatggtaga gcgcagatgc ccaccactga aagagtgact ttgacaagat gtttctacct
1561 cttcccagga catctcgagc accaccacca ccaccactga gatccggctg ctaacaaagc
1621 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
1681 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc
1741 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
1801 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
1861 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
1921 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
1981 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
2041 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
2101 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
2161 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
2221 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
2281 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
2341 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
2401 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
2461 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
2521 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
2581 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
2641 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
2701 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
2761 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
2821 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
2881 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
2941 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
3001 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
3061 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
3121 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
3181 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
3241 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
3301 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
3361 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
3421 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
3481 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
3541 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
3601 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
3661 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
3721 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
3781 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
3841 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
3901 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
3961 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
4021 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
4081 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
4141 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct
4201 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
4261 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
4321 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
4381 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
4441 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
4501 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
4561 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
4621 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
4681 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
4741 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag
4801 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga
4861 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg
4921 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg
4981 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggg
5041 gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg
5101 acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc
5161 gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt
5221 cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc
5281 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt
5341 gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg
5401 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
5461 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
5521 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
5581 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc
5641 gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg
5701 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
5761 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
5821 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
5881 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
5941 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
6001 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
6061 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
6121 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
6181 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
6241 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
6301 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
6361 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
6421 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
6481 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc
6541 cgggatctcg acgctctccc ttatgcgact cctgc
//
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