| 质粒名称: | pEVOL-AckRS |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 137976 |
| 存储实验室: | Wenshe Liu |
|
载体骨架基本信息 |
|
| 载体名称: | pEVOL |
| 载体出品公司: | -- |
| 载体抗性: | 氯霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 5651 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | AckRS and pylTcua |
| 物种来源: | Methanosarcina mazei |
| 插入大小: | 1365 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | araBAD |
| 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 | tttcagatgggttctggctgcacccgtg |
| 3' 测序引物 | aaagttcagcatagtgaactcttcc |
|
文献引用方法 |
|
| 材料和方法部分: | pEVOL-AckRS was a gift from Wenshe Liu (Addgene plasmid # 137976 ; http://n2t.net/addgene:137976 ; RRID:Addgene_137976) |
| 参考文献部分: | A Click Chemistry Approach Reveals the Chromatin-Dependent Histone H3K36 Deacylase Nature of SIRT7. Wang WW, Angulo-Ibanez M, Lyu J, Kurra Y, Tong Z, Wu B, Zhang L, Sharma V, Zhou J, Lin H, Gao YQ, Li W, Chua KF, Liu WR. J Am Chem Soc. 2019 Feb 13;141(6):2462-2473. doi: 10.1021/jacs.8b12083. Epub 2019 Feb 4. 10.1021/jacs.8b12083 PubMed 30653310 |
买家导航
pEVOLAckRS载体基本信息
pEVOLAckRS载体图谱质粒图谱和多克隆位点信息
pEVOLAckRS载体简介
pEVOLAckRS载体序列
LOCUS pEVOL-AckRS 5652 bp ds-DNA circular SYN 19-FEB-2020
DEFINITION sequence optimized N‐acetyl lysyl‐tRNA synthetase with cognate tRNA
for genetic code expansion.
ACCESSION .
VERSION .
KEYWORDS pEVOL-AckRS
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5652)
AUTHORS Wang WW, Angulo-Ibanez M, Lyu J, Kurra Y, Tong Z, Wu B, Zhang L,
Sharma V, Zhou J, Lin H, Gao YQ, Li W, Chua KF, Liu WR
TITLE A Click Chemistry Approach Reveals the Chromatin-Dependent Histone
H3K36 Deacylase Nature of SIRT7.
JOURNAL J Am Chem Soc. 2019 Feb 13;141(6):2462-2473. doi:
10.1021/jacs.8b12083. Epub 2019 Feb 4.
PUBMED 30653310
REFERENCE 2 (bases 1 to 5652)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Feb 19, 2020 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5652
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..285
/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 208..227
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
CDS 1693..1710
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(1766..1783)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
CDS complement(2769..3428)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 3343..3362
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(3429..3531)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
primer_bind complement(3923..3940)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4057..4602)
/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(4137..4156)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4748..5626)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPSDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
ORIGIN
1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg
301 ctaacaggag gaattactag tatggataag aaaccgctga atactctgat ttctgcaact
361 ggtctgtgga tgagccgtac cggcaccatc cacaagatca aacaccacga ggtttcccgt
421 agcaaaatct acatcgaaat ggcgtgcggt gaccacctgg tggtaaacaa ctcccgttct
481 tctcgtactg cacgtgctct gcgccaccac aagtaccgta agacctgcaa gcgctgtcgc
541 gtgtctgatg aagacctgaa caaattcctg actaaagcga acgaagatca gacttctgtg
601 aaggtgaaag ttgtttctgc cccaacccgc accaagaaag cgatgccgaa gtccgttgca
661 cgcgctccga aaccgctgga gaacaccgaa gccgcacagg cccagccgtc tggttctaag
721 ttttctccgg caatcccggt ttctactcag gagtctgtgt ctgtgccagc ttctgttagc
781 acttctattt cctctatcag cactggtgcg actgcgtccg ctctggtaaa aggtaacact
841 aacccgatca ccagcatgtc tgctccggtt caggcttctg caccggcact gactaaaagc
901 cagactgacc gtctggaggt tctgctgaac ccgaaagatg aaatcagcct gaactctggc
961 aaaccgttcc gtgaactgga atccgaactg ctgtctcgtc gtaagaaaga cctgcaacaa
1021 atctatgctg aagagcgtga aaactacctg ggtaaactgg aacgtgaaat cacccgtttc
1081 tttgtggacc gtggtttcct ggaaatcaag tctccgatcc tgatcccgct ggaatacatc
1141 gagcgcatgg gtattgataa cgacaccgaa ctgtccaagc agattttccg tgtggacaag
1201 aacttctgcc tgcgtccgat gatggcaccg aacctgctga attacgcgcg taaactggat
1261 cgtgcactgc cggacccgat caaaatcttt gaaatcggtc catgctatcg taaggagagc
1321 gacggtaaag aacacctgga agagttcact atgctgaact tttttcagat gggttctggc
1381 tgcacccgtg aaaatctgga atctatcatc accgacttcc tgaaccacct gggcattgac
1441 ttcaaaatcg ttggtgattc ctgcatggtt tacggtgaca ctctggacgt tatgcatggt
1501 gatctggaac tgagcagcgc tgttgtgggt ccgattccgc tggatcgtga atggggtatc
1561 gataaaccgt ggattggtgc tggcttcggt ctggaacgtc tgctgaaagt taagcacgac
1621 tttaagaaca tcaaacgtgc tgcgcgttcc gagtcctatt acaacggcat tagcactaac
1681 ctgtaagtcg accatcatca tcatcatcat tgagtttaaa cggtctccag cttggctgtt
1741 ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc
1801 tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga
1861 actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag
1921 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc cttgtttgtg
1981 agctcccggt catcaatcat ccccataatc cttgttagat tatcaatttt aaaaaactaa
2041 cagttgtcag cctgtcccgc tttaatatca tacgccgtta tacgttgttt acgctttgag
2101 gaatcccata tgatacgcgg ccgctttcaa acgctaaatt gcctgatgcg ctacgcttat
2161 caggcctaca tgatctctgc aatatattga gtttgcgtgc ttttgtaggc cggataaggc
2221 gttcacgccg catccggcaa gaaacagcaa acaatccaaa acgccgcgtt cagcggcgtt
2281 ttttctgctt ttcttcgcga attaattccg cttcgcaaca tgtgagcacc ggtttattga
2341 ctaccggaag cagtgtgacc gtgtgcttct caaatgcctg aggccagttt gctcaggctc
2401 tccccgtgga ggtaataatt gacgatatga tcagtgcacg gctaactaag cggcctgctg
2461 actttctcgc cgatcaaaag gcattttgct attaagggat tgacgagggc gtatctgcgc
2521 agtaagatgc gccccgcatt ggaaacctga tcatgtagat cgaatggact ctaaatccgt
2581 tcagccgggt tagattcccg gggtttccgc caaattcgaa aagcctgctc aacgagcagg
2641 cttttttgca tgctcgagca gctcagggtc gaatttgctt tcgaatttct gccattcatc
2701 cgcttattat cacttattca ggcgtagcaa ccaggcgttt aagggcacca ataactgcct
2761 taaaaaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt
2821 ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc
2881 accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc
2941 atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa
3001 aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca
3061 tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat
3121 gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc
3181 accagctcac cgtctttcat tgccatacgg aattccggat gagcattcat caggcgggca
3241 agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag
3301 gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc
3361 tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt
3421 ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt
3481 agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc aacgtctcat
3541 tttcgccaaa agttggccca gggcttcccg gtatcaacag ggacaccagg atttatttat
3601 tctgcgaagt gatcttccgt cacaggtatt tattcggcgc aaagtgcgtc gggtgatgct
3661 gccaacttac tgatttagtg tatgatggtg tttttgaggt gctccagtgg cttctgtttc
3721 tatcagctgt ccctcctgtt cagctactga cggggtggtg cgtaacggca aaagcaccgc
3781 cggacatcag cgctagcgga gtgtatactg gcttactatg ttggcactga tgagggtgtc
3841 agtgaagtgc ttcatgtggc aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt
3901 gatacaggat atattccgct tcctcgctca ctgactcgct acgctcggtc gttcgactgc
3961 ggcgagcgga aatggcttac gaacggggcg gagatttcct ggaagatgcc aggaagatac
4021 ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc cataggctcc gcccccctga
4081 caagcatcac gaaatctgac gctcaaatca gtggtggcga aacccgacag gactataaag
4141 ataccaggcg tttccccctg gcggctccct cgtgcgctct cctgttcctg cctttcggtt
4201 taccggtgtc attccgctgt tatggccgcg tttgtctcat tccacgcctg acactcagtt
4261 ccgggtaggc agttcgctcc aagctggact gtatgcacga accccccgtt cagtccgacc
4321 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac
4381 cactggcagc agccactggt aattgattta gaggagttag tcttgaagtc atgcgccggt
4441 taaggctaaa ctgaaaggac aagttttggt gactgcgctc ctccaagcca gttacctcgg
4501 ttcaaagagt tggtagctca gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg
4561 ttttcagagc aagagattac gcgcagacca aaacgatctc aagaagatca tcttattaat
4621 cagataaaat atttctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc
4681 agccccatac gatataagtt gtaattctca tgtttgacag cttatcatcg ataagcttgg
4741 tacccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa ccggcacgga
4801 actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag agttgatcgt
4861 caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc aaaagcagct
4921 tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct aactgctggc
4981 ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg ctggcgatat
5041 caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt acccgattat
5101 ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac aattgctcaa
5161 gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg ttaatgattt
5221 gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag aaccccgtat
5281 tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga cgaaagtaaa
5341 cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga atctctcctg
5401 gcgggaacag caaaatatca ctcggtcggc aaacaaattc tcgtccctga tttttcacca
5461 ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc ggtcggtcga
5521 taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc agatgggcat
5581 taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt ttcatactcc
5641 cgccattcag ag
//
