| 质粒名称: | pHyo094 (eMutaT7) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 173124 |
| 存储实验室: | Seokhee Kim |
|
载体骨架基本信息 |
|
| 载体名称: | pBAD33 |
| 载体出品公司: | -- |
| 空载体大小: | 5340 bp |
| 完整质粒大小: | 8947 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 氯霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | pmCDA1-T7RNAP |
| 物种来源: | Synthetic |
| 插入大小: | 3607 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | pBAD-F (5'-ATGCCATAGCATTTTTATCC-3') |
| 3’测序引物 | pBAD-R (5'-GATTTAATCTGTATCAGG-3') |
|
文献引用方法 |
|
| 材料和方法部分: | pHyo094 (eMutaT7) was a gift from Seokhee Kim (Addgene plasmid # 173124 ; http://n2t.net/addgene:173124 ; RRID:Addgene_173124) |
买家导航
pHyo094 (eMutaT7)载体质粒基本信息
pHyo094 (eMutaT7)质粒图谱载体图谱和pHyo094 (eMutaT7)载体序列质粒序列多克隆位点信息
质粒图谱.png)
pHyo094 (eMutaT7)质粒载体简介
pHyo094 (eMutaT7)质粒序列载体序列
LOCUS pHyo094 (eMutaT7) 8956 bp ds-DNA circular SYN 14-SEP-2021
DEFINITION Expresses eMutaT7 for mutagenizing genes downstream from a T7
promoter in e.coli.
ACCESSION .
VERSION .
KEYWORDS pHyo094 (eMutaT7)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8956)
AUTHORS Park H, Kim S
TITLE Gene-specific mutagenesis enables rapid continuous evolution of
enzymes in vivo.
JOURNAL Nucleic Acids Res. 2021 Apr 6;49(6):e32. doi: 10.1093/nar/gkaa1231.
PUBMED 33406230
REFERENCE 2 (bases 1 to 8956)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 14, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8956
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 43..70
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 89..180
/gene="bla"
/label=AmpR promoter
rep_origin 638..1093
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(725..744)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 935..956
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(1649..2308)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 2223..2242
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(2309..2411)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
primer_bind complement(2803..2820)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2937..3482)
/direction=LEFT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin.
"
primer_bind complement(3017..3036)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3558..3577
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
CDS complement(3757..4635)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
promoter 4662..4946
/gene="araBAD"
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
primer_bind 4869..4888
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
CDS 8358..8612
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHT
AYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
primer_bind complement(8651..8668)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 8821..8907
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
ORIGIN
1 gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg
61 atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca ttcaaatatg
121 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
181 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
241 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
301 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
361 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
421 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
481 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
541 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
601 cagaccaagt ttactcatat atactttaga ttgatttacg cgccctgtag cggcgcatta
661 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
721 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
781 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
841 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
901 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
961 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc
1021 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
1081 acgtttacaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
1141 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
1201 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
1261 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
1321 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
1381 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtcag
1441 gcatttgaga agcacacggt cacactgctt ccggtagtca ataaaccggt aaaccagcaa
1501 tagacataag cggctattta acgaccctgc cctgaaccga cgaccgggtc gaatttgctt
1561 tcgaatttct gccattcatc cgcttattat cacttattca ggcgtagcaa ccaggcgttt
1621 aagggcacca ataactgcct taaaaaaatt acgccccgcc ctgccactca tcgcagtact
1681 gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca tgatgaacct
1741 gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc atggtgaaaa
1801 cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc
1861 agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa taggccaggt
1921 tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt
1981 ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag
2041 ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacgg aattccggat
2101 gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt
2161 tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt
2221 gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg
2281 tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa aatctcgata
2341 actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta
2401 cgtgccgatc aacgtctcat tttcgccaaa agttggccca gggcttcccg gtatcaacag
2461 ggacaccagg atttatttat tctgcgaagt gatcttccgt cacaggtatt tattcggcgc
2521 aaagtgcgtc gggtgatgct gccaacttac tgatttagtg tatgatggtg tttttgaggt
2581 gctccagtgg cttctgtttc tatcagctgt ccctcctgtt cagctactga cggggtggtg
2641 cgtaacggca aaagcaccgc cggacatcag cgctagcgga gtgtatactg gcttactatg
2701 ttggcactga tgagggtgtc agtgaagtgc ttcatgtggc aggagaaaaa aggctgcacc
2761 ggtgcgtcag cagaatatgt gatacaggat atattccgct tcctcgctca ctgactcgct
2821 acgctcggtc gttcgactgc ggcgagcgga aatggcttac gaacggggcg gagatttcct
2881 ggaagatgcc aggaagatac ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc
2941 cataggctcc gcccccctga caagcatcac gaaatctgac gctcaaatca gtggtggcga
3001 aacccgacag gactataaag ataccaggcg tttccccctg gcggctccct cgtgcgctct
3061 cctgttcctg cctttcggtt taccggtgtc attccgctgt tatggccgcg tttgtctcat
3121 tccacgcctg acactcagtt ccgggtaggc agttcgctcc aagctggact gtatgcacga
3181 accccccgtt cagtccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
3241 ggaaagacat gcaaaagcac cactggcagc agccactggt aattgattta gaggagttag
3301 tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac aagttttggt gactgcgctc
3361 ctccaagcca gttacctcgg ttcaaagagt tggtagctca gagaaccttc gaaaaaccgc
3421 cctgcaaggc ggttttttcg ttttcagagc aagagattac gcgcagacca aaacgatctc
3481 aagaagatca tcttattaat cagataaaat atttctaggc tcatgagccc gaagtggcga
3541 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg
3601 ccggtgatgc cggccacgat gcgtccggcg tagaggatct gctcatgttt gacagcttat
3661 catcgatgca taatgtgcct gtcaaatgga cgaagcaggg attctgcaaa ccctatgcta
3721 ctccgtcaag ccgtcaattg tctgattcgt taccaattat gacaacttga cggctacatc
3781 attcactttt tcttcacaac cggcacggaa ctcgctcggg ctggccccgg tgcatttttt
3841 aaatacccgc gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat
3901 aggcatccgg gtggtgctca aaagcagctt cgcctggctg atacgttggt cctcgcgcca
3961 gcttaagacg ctaatcccta actgctggcg gaaaagatgt gacagacgcg acggcgacaa
4021 gcaaacatgc tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt gatcgctgat
4081 gtactgacaa gcctcgcgta cccgattatc catcggtgga tggagcgact cgttaatcgc
4141 ttccatgcgc cgcagtaaca attgctcaag cagatttatc gccagcagct ccgaatagcg
4201 cccttcccct tgcccggcgt taatgatttg cccaaacagg tcgctgaaat gcggctggtg
4261 cgcttcatcc gggcgaaaga accccgtatt ggcaaatatt gacggccagt taagccattc
4321 atgccagtag gcgcgcggac gaaagtaaac ccactggtga taccattcgc gagcctccgg
4381 atgacgaccg tagtgatgaa tctctcctgg cgggaacagc aaaatatcac ccggtcggca
4441 aacaaattct cgtccctgat ttttcaccac cccctgaccg cgaatggtga gattgagaat
4501 ataacctttc attcccagcg gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat
4561 cggcgttaaa cccgccacca gatgggcatt aaacgagtat cccggcagca ggggatcatt
4621 ttgcgcttca gccatacttt tcatactccc gccattcaga gaagaaacca attgtccata
4681 ttgcatcaga cattgccgtc actgcgtctt ttactggctc ttctcgctaa ccaaaccggt
4741 aaccccgctt attaaaagca ttctgtaaca aagcgggacc aaagccatga caaaaacgcg
4801 taacaaaagt gtctataatc acggcagaaa agtccacatt gattatttgc acggcgtcac
4861 actttgctat gccatagcat ttttatccat aagattagcg gatcctacct gacgcttttt
4921 atcgcaactc tctactgttt ctccataccc gtttttttgg gctagcgaat tcgagctcag
4981 gaggaattca ccatggcatt atcatatgat gacagacgcc gagtacgtgc gcattcatga
5041 gaaactggat atttacacct tcaagaagca gttcttcaac aacaagaaat ctgtgtcaca
5101 ccgctgctac gtgctgtttg agttgaagcg aaggggcgaa agaagggctt gcttttgggg
5161 ctatgccgtc aacaagcccc aaagtggcac cgagagagga atacacgctg agatattcag
5221 tatccgaaag gtggaagagt atcttcggga taatcctggg cagtttacga tcaactggta
5281 ttccagctgg agtccttgcg ctgattgtgc cgagaaaatt ctggaatggt ataatcagga
5341 acttcgggga aacgggcaca cattgaaaat ctgggcctgc aagctgtact acgagaagaa
5401 tgcccggaac cagataggac tctggaatct gagggacaat ggtgtaggcc tgaacgtgat
5461 ggtttccgag cactatcagt gttgtcggaa gattttcatc caaagctctc ataaccagct
5521 caatgaaaac cgctggttgg agaaaacact gaaacgtgcg gagaagtgga gatccgagct
5581 gagcatcatg atccaggtca agattctgca taccactaag tctccagccg ttagcgagac
5641 tcccgggacc tcagagtccg ccacacccga aagtaacacg attaacatcg ctaagaacga
5701 cttctctgac atcgaactgg ctgctatccc gttcaacact ctggctgacc attacggtga
5761 gcgtttagct cgcgaacagt tggcccttga gcatgagtct tacgagatgg gtgaagcacg
5821 cttccgcaag atgtttgagc gtcaacttaa agctggtgag gttgcggata acgctgccgc
5881 caagcctctc atcactaccc tactccctaa gatgattgca cgcatcaacg actggtttga
5941 ggaagtgaaa gctaagcgcg gcaagcgccc gacagccttc cagttcctgc aagaaatcaa
6001 gccggaagcc gtagcgtaca tcaccattaa gaccactctg gcttgcctaa ccagtgctga
6061 caatacaacc gttcaggctg tagcaagcgc aatcggtcgg gccattgagg acgaggctcg
6121 cttcggtcgt atccgtgacc ttgaagctaa gcacttcaag aaaaacgttg aggaacaact
6181 caacaagcgc gtagggcacg tctacaagaa agcatttatg caagttgtcg aggctgacat
6241 gctctctaag ggtctactcg gtggcgaggc gtggtcttcg tggcataagg aagactctat
6301 tcatgtagga gtacgctgca tcgagatgct cattgagtca accggaatgg ttagcttaca
6361 ccgccaaaat gctggcgtag taggtcaaga ctctgagact atcgaactcg cacctgaata
6421 cgctgaggct atcgcaaccc gtgcaggtgc gctggctggc atctctccga tgttccaacc
6481 ttgcgtagtt cctcctaagc cgtggactgg cattactggt ggtggctatt gggctaacgg
6541 tcgtcgtcct ctggcgctgg tgcgtactca cagtaagaaa gcactgatgc gctacgaaga
6601 cgtttacatg cctgaggtgt acaaagcgat taacattgcg caaaacaccg catggaaaat
6661 caacaagaaa gtcctagcgg tcgccaacgt aatcaccaag tggaagcatt gtccggtcga
6721 ggacatccct gcgattgagc gtgaagaact cccgatgaaa ccggaagaca tcgacatgaa
6781 tcctgaggct ctcaccgcgt ggaaacgtgc tgccgctgct gtgtaccgca aggacaaggc
6841 tcgcaagtct cgccgtatca gccttgagtt catgcttgag caagccaata agtttgctaa
6901 ccataaggcc atctggttcc cttacaacat ggactggcgc ggtcgtgttt acgctgtgtc
6961 aatgttcaac ccgcaaggta acgatatgac caaaggactg cttacgctgg cgaaaggtaa
7021 accaatcggt aaggaaggtt actactggct gaaaatccac ggtgcaaact gtgcgggtgt
7081 cgataaggtt ccgttccctg agcgcatcaa gttcattgag gaaaaccacg agaacatcat
7141 ggcttgcgct aagtctccac tggagaacac ttggtgggct gagcaagatt ctccgttctg
7201 cttccttgcg ttctgctttg agtacgctgg ggtacagcac cacggcctga gctataactg
7261 ctcccttccg ctggcgtttg acgggtcttg ctctggcatc cagcacttct ccgcgatgct
7321 ccgagatgag gtaggtggtc gcgcggttaa cttgcttcct agtgaaaccg ttcaggacat
7381 ctacgggatt gttgctaaga aagtcaacga gattctacaa gcagacgcaa tcaatgggac
7441 cgataacgaa gtagttaccg tgaccgatga gaacactggt gaaatctctg agaaagtcaa
7501 gctgggcact aaggcactgg ctggtcaatg gctggcttac ggtgttactc gcagtgtgac
7561 taagcgttca gtcatgacgc tggcttacgg gtccaaagag ttcggcttcc gtcaacaagt
7621 gctggaagat accattcagc cagctattga ttccggcaag ggtctgatgt tcactcagcc
7681 gaatcaggct gctggataca tggctaagct gatttgggaa tctgtgagcg tgacggtggt
7741 agctgcggtt gaagcaatga actggcttaa gtctgctgct aagctgctgg ctgctgaggt
7801 caaagataag aagactggag agattcttcg caagcgttgc gctgtgcatt gggtaactcc
7861 tgatggtttc cctgtgtggc aggaatacaa gaagcctatt cagacgcgct tgaacctgat
7921 gttcctcggt cagttccgct tacagcctac cattaacacc aacaaagata gcgagattga
7981 tgcacacaaa caggagtctg gtatcgctcc taactttgta cacagccaag acggtagcca
8041 ccttcgtaag actgtagtgt gggcacacga gaagtacgga atcgaatctt ttgcactgat
8101 tcacgactcc ttcggtacca ttccggctga cgctgcgaac ctgttcaaag cagtgcgcga
8161 aactatggtt gacacctatg agtcttgtga tgtactggct gatttctacg accagttcgc
8221 tgaccagttg cacgagtctc aattggacaa aatgccagca cttccggcta aaggtaactt
8281 gaacctccgt gacatcttag agtcggactt cgcgttcgcg taactgcagc ccggggatcc
8341 tctagagtcg acctgcgatg accaaccttt ccgacatcat agagaaggaa acaggcaaac
8401 agttggtcat ccaagagtcg atactcatgc ttcctgaaga agttgaggag gtcattggga
8461 ataagccgga aagtgacatt ctcgtacaca ctgcgtatga tgagagcacc gatgagaacg
8521 tgatgctgct cacgtcagat gccccagagt acaaaccctg ggctctggtg attcaggact
8581 ctaatggaga gaacaagatc aagatgctgt aaaagcttgg ctgttttggc ggatgagaga
8641 agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata aaacagaatt
8701 tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac
8761 gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac tgccaggcat
8821 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
8881 gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa
8941 cggcccggag ggtggc
//
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