| 质粒名称: | pmal_c5x_RNAse_Inhib |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 153314 |
| 存储实验室: | Drew Endy, Philippa Marrack |
|
载体骨架基本信息 |
|
| 载体名称: | pMAL-c5X |
| 载体出品公司: | NEB |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 30 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | RNAse Inhibitor |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | Maltose Binding Protein（主链N端） |
| 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: | pmal_c5x_RNAse_Inhib was a gift from Drew Endy & Philippa Marrack (Addgene plasmid # 153314 ; http://n2t.net/addgene:153314 ; RRID:Addgene_153314) |
买家导航
pmal_c5x_RNAse_Inhib载体质粒基本信息
pmal_c5x_RNAse_Inhib质粒图谱载体图谱和pmal_c5x_RNAse_Inhib载体序列质粒序列多克隆位点信息
pmal_c5x_RNAse_Inhib质粒载体简介
pmal_c5x_RNAse_Inhib质粒序列载体序列
LOCUS pmal_c5x_RNAse_Inhib 6994 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Bacterial expression of an RNase inhibitor.
ACCESSION .
VERSION .
KEYWORDS pmal_c5x_RNAse_Inhib
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6994)
TITLE COVID-19 Diagnostic Toolkit Enzymes
REFERENCE 2 (bases 1 to 6994)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6994
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 152..180
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 188..204
/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 193..215
/label=M13/pUC Reverse
/note="In lacZ gene"
CDS 274..1374
/codon_start=1
/gene="malE (mutated)"
/product="maltose binding protein from E. coli"
/label=MBP
/note="This version of the gene does not encode a signal
sequence, so MBP will remain in the cytosol."
/translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
ENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPQMSAF
WYAVRTAVINAASGRQTVDEALKDAQT"
primer_bind 1348..1371
/label=MBP-F
/note="Maltose binding protein, forward primer"
CDS 1423..1434
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
terminator 2826..2912
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 3004..3031
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 3059..3149
/gene="bla"
/label=AmpR promoter
CDS 3150..4010
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(3368..3387)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4098..4686
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4587..4606
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4840..4857
/label=L4440
/note="L4440 vector, forward primer"
primer_bind 5040..5062
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(5056..5247)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
promoter 5743..5820
/gene="lacI (mutant)"
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
CDS 5821..6903
/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(5840..5859)
/label=LacI-R
/note="LacI, reverse primer"
ORIGIN
1 ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat
61 aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata
121 acggttctgg caaatattct gaaatgagct gttgacaatt aatcatcggc tcgtataatg
181 tgtggaattg tgagcggata acaatttcac acaggaaaca gccagtccgt ttaggtgttt
241 tcacgagcaa ttgaccaaca aggaccatag attatgaaaa tcgaagaagg taaactggta
301 atctggatta acggcgataa aggctataac ggtctcgctg aagtcggtaa gaaattcgag
361 aaagataccg gaattaaagt caccgttgag catccggata aactggaaga gaaattccca
421 caggttgcgg caactggcga tggccctgac attatcttct gggcacacga ccgctttggt
481 ggctacgctc aatctggcct gttggctgaa atcaccccgg acaaagcgtt ccaggacaag
541 ctgtatccgt ttacctggga tgccgtacgt tacaacggca agctgattgc ttacccgatc
601 gctgttgaag cgttatcgct gatttataac aaagatctgc tgccgaaccc gccaaaaacc
661 tgggaagaga tcccggcgct ggataaagaa ctgaaagcga aaggtaagag cgcgctgatg
721 ttcaacctgc aagaaccgta cttcacctgg ccgctgattg ctgctgacgg gggttatgcg
781 ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa cgctggcgcg
841 aaagcgggtc tgaccttcct ggttgacctg attaaaaaca aacacatgaa tgcagacacc
901 gattactcca tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac catcaacggc
961 ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt atggtgtaac ggtactgccg
1021 accttcaagg gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg tattaacgcc
1081 gccagtccga acaaagagct ggcaaaagag ttcctcgaaa actatctgct gactgatgaa
1141 ggtctggaag cggttaataa agacaaaccg ctgggtgccg tagcgctgaa gtcttacgag
1201 gaagagttgg tgaaagatcc gcgtattgcc gccactatgg aaaacgccca gaaaggtgaa
1261 atcatgccga acatcccgca gatgtccgct ttctggtatg ccgtgcgtac tgcggtgatc
1321 aacgccgcca gcggtcgtca gactgtcgat gaagccctga aagacgcgca gactaattcg
1381 agctcgaaca acaacaacaa taacaataac aacaacctcg ggattgaggg aaggagtctt
1441 gacatccagt gtgagcagct gagtgatgcc cggtggacag agctccttcc tctgatccag
1501 caatacgaag tggtcaggct ggatgactgt ggcctcactg aagtgcggtg caaagacatc
1561 agctcagcag tccaagccaa ccctgccctg acagagctca gccttcgcac caatgaactg
1621 ggtgatggcg gtgtgggtct ggtgctccag ggcctgcaga atcccacttg taagatccag
1681 aagctgagcc ttcagaactg cggcttgacg gaggctggct gtgggatcct gcctggcatg
1741 ctgcgctctt tgtctaccct gcgtgagcta catctcaatg ataaccctat gggggacgca
1801 ggcctgaagc tgctctgtga aggacttcag gacccccagt gccgtcttga gaagcttcag
1861 ttggaatact gtaacctcac agctaccagc tgcgagcccc tggcttcagt gctcagggtg
1921 aaagctgact ttaaggagct agtattgagc aacaacgacc tccatgaacc tggtgtccgt
1981 attctgtgcc agggcctgaa ggactctgcc tgtcaactgg agtcactcaa gctggagaac
2041 tgtggtatca cagcagccaa ctgcaaggat ctgtgcgatg ttgtggcctc caaagcttcg
2101 ctacaggaac tggacttgag cagcaacaag ctaggtaatg caggcattgc agcactgtgc
2161 ccaggactgc tgcttcccag ctgtaagctc aggactctgt ggctctggga atgtgatatc
2221 actgcagagg gctgcaagga cctgtgccgt gtcctcagag ccaagcagag cctgaaggaa
2281 ctcagccttg cttccaatga gctgaaggat gagggtgccc ggctgctgtg tgagagccta
2341 ttagagcctg gctgtcagct ggagtcactg tggataaaga cctgtagcct cacagctgcc
2401 tcctgtccct acttctgctc cgtgttgacc aaaagccgtt ctctgttgga gctgcaaatg
2461 agcagcaacc cactgggaga cgagggagtc caggagcttt gcaaggccct gagccagcca
2521 gatacagtgc tgcgtgagct ttggttggga gactgtgatg tgacaaatag tggttgcagc
2581 agccttgcca atgtcctgct ggccaaccgc agcctgaggg aactggacct cagtaacaac
2641 tgcatggggg gcccaggtgt cctgcaactg ctggagagcc tcaaacagcc cagctgcacc
2701 cttcagcagc tcgtcctgta tgacatttac tggacaaatg aggtggaaga acagcttcgg
2761 gccctggagg aggaaaggcc ttccctgagg atcatttcct aacctgcagg taattaaata
2821 agcttcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
2881 tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga
2941 agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta
3001 agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt tcggtccgtt
3061 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
3121 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
3181 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
3241 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
3301 gcggtaagat ccttgagagt tttcgccccg aagaacgttt cccaatgatg agcactttta
3361 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
3421 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
3481 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
3541 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
3601 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
3661 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
3721 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
3781 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
3841 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
3901 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
3961 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
4021 aagtttactc atatatactt tagattgatt tccttaggac tgagcgtcaa ccccgtagaa
4081 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
4141 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
4201 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
4261 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
4321 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
4381 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
4441 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
4501 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
4561 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
4621 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
4681 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
4741 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
4801 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
4861 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
4921 atataaggtg cactgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
4981 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
5041 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
5101 gtaaagctca tcagcgtggt cgtgcagcga ttcacagatg tctgcctgtt catccgcgtc
5161 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
5221 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
5281 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
5341 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
5401 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc
5461 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga
5521 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca
5581 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt
5641 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat
5701 catgcgcacc cgtggccagg acccaacgct gcccgaaatt ccgacaccat cgaatggtgc
5761 aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat
5821 gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt
5881 tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg
5941 gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag
6001 tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc
6061 gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa
6121 cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt
6181 gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc
6241 actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt
6301 ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag
6361 caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc
6421 tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg
6481 agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact
6541 gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc
6601 gggctgcgcg ttggtgcgga tatttcggta gtgggatacg acgataccga agacagctca
6661 tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc
6721 gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc
6781 gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cacaaaccgc ctctccccgc
6841 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
6901 tgagcgcaac gcaattaatg taagttagct cactcattag gcacaattct catgtttgac
6961 agcttatcat cgactgcacg gtgcaccaat gctt
//
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