| 质粒名称: | pET-21a(+)-IPP1-His |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 118978 |
| 存储实验室: | Sebastian Maerkl |
|
载体骨架基本信息 |
|
| 载体名称: | pET21a(+) |
| 载体出品公司: | EMD Biosciences |
| 空载体大小: | 5443 bp |
| 完整质粒大小: | 6099 bp |
| 载体修饰: | 含有T7启动子、RBS和T7终止子的部分原始骨架被插入物取代。 |
| 载体类型: | 细菌表达 |
| 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: | inorganic diphosphatase IPP1,Inorganic pyrophosphatase |
| 物种来源: | S. cerevisiae (budding yeast) |
| 插入大小: | 1376 bp |
| GenBank编号 | NM_001178359.1 |
| 启动子: | T7 promoter |
| 标签/融合蛋白: | 6x His（插入件上的C端子） |
| 克隆方法: | Gibson克隆 |
| 5’测序引物 | GCGTCCCATTCGCCAATCCG |
| 3’测序引物 | GCGTCCGGCGTAGAGGATC |
|
文献引用方法 |
|
| 材料和方法部分: | pET-21a(+)-IPP1-His was a gift from Sebastian Maerkl (Addgene plasmid # 118978 ; http://n2t.net/addgene:118978 ; RRID:Addgene_118978) |
| 参考文献部分: | A simple, robust, and low-cost method to produce the PURE cell-free system. Lavickova B, Maerkl SJ. ACS Synth Biol. 2019 Jan 11. doi: 10.1021/acssynbio.8b00427. 10.1021/acssynbio.8b00427 PubMed 30632751 |
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
-IPP1-His载体图谱.png)
载体简介
载体序列
LOCUS pET-21a(+)-IPP1-His 6228 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Bacterial expression plasmid for Inorganic pyrophosphatase.
ACCESSION .
VERSION .
KEYWORDS pET-21a(+)-IPP1-His
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6228)
AUTHORS Lavickova B, Maerkl SJ
TITLE A simple, robust, and low-cost method to produce the PURE cell-free
system.
JOURNAL ACS Synth Biol. 2019 Jan 11. doi: 10.1021/acssynbio.8b00427.
PUBMED 30632751
REFERENCE 2 (bases 1 to 6228)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6228
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 970..989
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(1009..1086)
/gene="lacI"
/label=lacI promoter
primer_bind 1292..1311
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 1399..1418
/label=T7
/note="T7 promoter, forward primer"
promoter 1399..1417
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 1418..1442
/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 1457..1479
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 2354..2371
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(2424..2442)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 2438..2485
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 2522..2977
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2609..2628)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2819..2840
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 3003..3107
/gene="bla"
/label=AmpR promoter
CDS 3108..3968
/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(3326..3345)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4139..4727
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4628..4647
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4881..4898
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 4913..5052
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(4978..4997)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5138..5160
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(5154..5345)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(6154..1008)
/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 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
61 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
121 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
181 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
241 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
301 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
361 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
421 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
481 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
541 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
601 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
661 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
721 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
781 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
841 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
901 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
961 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
1021 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
1081 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
1141 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
1201 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
1261 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
1321 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatcg
1381 atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt
1441 cccctctaga aataattttg tttaacttta agaaggagat atacatatga cctacactac
1501 cagacaaatt ggtgccaaga acaccttgga atacaaagtt tatatcgaaa aggatggtaa
1561 gccagtttct gccttccacg acattccctt gtacgctgac aaggaaaaca acattttcaa
1621 catggttgtt gaaattccac gttggaccaa cgccaagtta gaaatcacca aggaagaaac
1681 tttgaaccca atcatccaag acaccaagaa gggcaagttg agatttgtta gaaactgttt
1741 ccctcaccat ggttacattc acaactatgg tgctttccca caaacttggg aagacccaaa
1801 cgtaagccac ccagaaacta aggcagttgg tgacaacgat ccaattgatg tgttggaaat
1861 tggtgaaact attgcttaca ctggtcaagt caagcaagtt aaggctctag gtatcatggc
1921 tttattggat gaaggtgaga ccgattggaa agttattgcc attgatatta acgatccatt
1981 agccccaaaa ttgaacgaca ttgaggatgt tgagaaatac ttcccaggtc tgttgagggc
2041 tactaacgaa tggttcagaa tttacaaaat cccagatggt aagccagaaa accaatttgc
2101 cttctccggt gaagctaaga acaagaagta cgctttggat atcatcaagg aaacacatga
2161 ctcctggaaa caattaattg ctggtaagtc ttctgacagc aagggtattg atttgaccaa
2221 tgttactttg cctgacaccc caacctactc caaggctgcc tctgatgcca tcccaccagc
2281 ttctccaaag gcagatgctc caattgacaa gtctattgac aagtggttct tcatctccgg
2341 ttctgttctc gagcaccacc accaccacca ctgagatccg gctgctaaca aagcccgaaa
2401 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
2461 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg
2521 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
2581 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
2641 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
2701 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
2761 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
2821 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
2881 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
2941 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
3001 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
3061 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
3121 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
3181 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
3241 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
3301 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
3361 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
3421 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
3481 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
3541 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
3601 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
3661 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
3721 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
3781 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
3841 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
3901 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
3961 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
4021 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
4081 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
4141 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
4201 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
4261 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
4321 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
4381 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
4441 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
4501 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
4561 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
4621 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
4681 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
4741 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
4801 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
4861 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
4921 ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac
4981 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
5041 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
5101 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
5161 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
5221 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
5281 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
5341 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
5401 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
5461 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
5521 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
5581 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
5641 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
5701 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
5761 cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg tggggccgcc
5821 atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag
5881 gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg
5941 ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg
6001 agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac
6061 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa
6121 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
6181 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggc
//
载体图谱质粒图谱pdf版和相关资料下载
载体质粒应用举例
- 上一篇：phage UbiC G3BP1-SNAP
- 下一篇：CoV2-M-IRES-E
