| 质粒名称: | pDule-pCNF |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 85494 |
| 存储实验室: | Ryan Mehl |
|
载体骨架基本信息 |
|
| 载体名称: | pDule |
| 载体出品公司: | -- |
| 载体抗性: | 四环素 |
| 空载体大小: | 5400 bp |
| 完整质粒大小: | 6300 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | para-cyanophenylalanine Mj synthetase,pCNF,pCNPhe,p-azidoPhe synthetase |
| 物种来源: | Methanocaldococcus jannaschii |
| 插入大小: | 921 bp |
| 基因突变: | Y32L L65V F108W Q109M D158G I159P |
| GenBank编号 | -- |
| 启动子 | lpp (constitutive) |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NcoI(未破坏) |
| 3' 克隆位点: | KpnI (未破坏) |
| 5' 测序引物 | CGTCACTGCGTCTTTTACTG |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pDule-pCNF was a gift from Ryan Mehl (Addgene plasmid # 85494 ; http://n2t.net/addgene:85494 ; RRID:Addgene_85494) |
| 参考文献部分: | Probing protein folding using site-specifically encoded unnatural amino acids as FRET donors with tryptophan. Miyake-Stoner SJ, Miller AM, Hammill JT, Peeler JC, Hess KR, Mehl RA, Brewer SH. Biochemistry. 2009 Jun 30;48(25):5953-62. doi: 10.1021/bi900426d. 10.1021/bi900426d PubMed 19492814 |
买家导航
pDulepCNF载体基本信息
pDulepCNF载体图谱质粒图谱和多克隆位点信息
pDulepCNF载体简介
pDulepCNF载体序列
LOCUS pDule-pCNF 6333 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Machinery plasmid expressing permissive pCNF synthetase and cognate
amber suppressing tRNA. Incorporates azidoPhenylalanine..
ACCESSION .
VERSION .
KEYWORDS pDule-pCNF
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6333)
AUTHORS Miyake-Stoner SJ, Miller AM, Hammill JT, Peeler JC, Hess KR, Mehl
RA, Brewer SH
TITLE Probing protein folding using site-specifically encoded unnatural
amino acids as FRET donors with tryptophan.
JOURNAL Biochemistry. 2009 Jun 30;48(25):5953-62. doi: 10.1021/bi900426d.
PUBMED 19492814
REFERENCE 2 (bases 1 to 6333)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6333
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 1161..1190
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1206..1223
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(1279..1296)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 1449..1535
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind complement(1742..1759)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1876..2421)
/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(1956..1975)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 2533..2561
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
CDS 2609..3799
/codon_start=1
/gene="tet"
/product="tetracycline efflux protein"
/label=TcR
/note="confers resistance to tetracycline"
/translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
primer_bind complement(2961..2980)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 4106..4128
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 4120..4137
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 4121..4137
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 5121..5143
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(5299..6177)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
ORIGIN
1 aactcagaat aagaaatgag gccgctcatg gcgttctgtt gcccgtctca ccggtgaaaa
61 gaaaaacaac cctggcgccg cttctttgag cgaacgatca aaaataagtg gcgccccatc
121 aaaaaaatat tctcaacata aaaaactttg tgtaatactt gtaacgctga gtttacgctt
181 tgaggaatcc cccatggacg aatttgaaat gataaagaga aacacatctg aaattatcag
241 cgaggaagag ttaagagagg ttttaaaaaa agatgaaaaa tctgctctga taggttttga
301 accaagtggt aaaatacatt tagggcatta tctccaaata aaaaagatga ttgatttaca
361 aaatgctgga tttgatataa ttatagtgtt ggctgattta catgcctatt taaaccagaa
421 aggagagttg gatgagatta gaaaaatagg agattataac aaaaaagttt ttgaagcaat
481 ggggttaaag gcaaaatatg tttatggaag tgaatggatg cttgataagg attatacact
541 gaatgtctat agattggctt taaaaactac cttaaaaaga gcaagaagga gtatggaact
601 tatagcaaga gaggatgaaa atccaaaggt tgctgaagtt atctatccaa taatgcaggt
661 taatggtcct cattatcttg gcgttgatgt tgcagttgga gggatggagc agagaaaaat
721 acacatgtta gcaagggagc ttttaccaaa aaaggttgtt tgtattcaca accctgtctt
781 aacgggtttg gatggagaag gaaagatgag ttcttcaaaa gggaatttta tagctgttga
841 tgactctcca gaagagatta gggctaagat aaagaaagca tactgcccag ctggagttgt
901 tgaaggaaat ccaataatgg agatagctaa atacttcctt gaatatcctt taaccataaa
961 aaggccagaa aaatttggtg gagatttgac agttaatagc tatgaggagt tagagagttt
1021 atttaaaaat aaggaattgc atccaatgga tttaaaaaat gctgtagctg aagaacttat
1081 aaagatttta gagccaatta gaaagagatt ataactgcag tttcaaacgg gtaccatatg
1141 ggaattcgaa gcttgggccc gaacaaaaac tcatctcaga agaggatctg aatagcgccg
1201 tcgaccatca tcatcatcat cattgagttt aaacggtctc cagcttggct gttttggcgg
1261 atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa
1321 acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga
1381 agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg
1441 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
1501 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagctgtc cctcctgttc
1561 agctactgac ggggtggtgc gtaacggcaa aagcaccgcc ggacatcagc gctagcggag
1621 tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct tcatgtggca
1681 ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata tattccgctt
1741 cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg
1801 aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa gtgagagggc
1861 cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg aaatctgacg
1921 ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
1981 cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca ttccgctgtt
2041 atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca gttcgctcca
2101 agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta tccggtaact
2161 atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca gccactggta
2221 attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca
2281 agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt ggtagctcag
2341 agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca agagattacg
2401 cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata tttctagatt
2461 tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg
2521 taattctcat gtttgacagc ttatcatcga taagctttaa tgcggtagtt tatcacagtt
2581 aaattgctaa cgcagtcagg caccgtgtat gaaatctaac aatgcgctca tcgtcatcct
2641 cggcaccgtc accctggatg ctgtaggcat aggcttggtt atgccggtac tgccgggcct
2701 cttgcgggat atcgtccatt ccgacagcat cgccagtcac tatggcgtgc tgctagcgct
2761 atatgcgttg atgcaatttc tatgcgcacc cgttctcgga gcactgtccg accgctttgg
2821 ccgccgccca gtcctgctcg cttcgctact tggagccact atcgactacg cgatcatggc
2881 gaccacaccc gtcctgtgga tcctctacgc cggacgcatc gtggccggca tcaccggcgc
2941 cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg
3001 ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg
3061 gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg tgctcaacgg
3121 cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag agcgtcgacc
3181 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat
3241 cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc
3301 gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc
3361 gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac
3421 caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccgacg cgctgggcta
3481 cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc cccattatga ttcttctcgc
3541 ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg tagatgacga
3601 ccatcaggga cagcttcaag gatcgctcgc ggctcttacc agcctaactt cgatcattgg
3661 accgctgatc gtcacggcga tttatgccgc ctcggcgagc acatggaacg ggttggcatg
3721 gattgtaggc gccgccctat accttgtctg cctccccgcg ttgcgtcgcg gtgcatggag
3781 ccgggccacc tcgacctgaa tggaagccgg cggcacctcg ctaacggatt caccactcca
3841 agaattggag ccaatcaatt cttgcggaga actgtgaatg cgcaaaccaa cccttggcag
3901 aacatatcca tcgcgtccgc catctccagc agccgcacgc ggcgcatctc gggctccttg
3961 catgcaccat tccttgcggc ggcggtgctc aacggcctca acctactact gggctgcttc
4021 ctaatgcagg agtcgcataa gggagagcgt ctggcgaaag ggggatgtgc tgcaaggcga
4081 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc
4141 aagcttaaaa aaaatcctta gctttcgcta aggatctgca gtggtccggc gggccggatt
4201 tgaaccagcg ccatgcggat ttagagtccg ccgttctgcc ctgctgaact accgccggga
4261 attcagcgtt acaagtatta cacaaagttt tttatgttga gaatattttt ttgatggggc
4321 gccacttatt tttgatcgtt cgctcaaaga agcggcgcca gggttgtttt tcttttcacc
4381 agtgagacgg gcaacagaac gccatgagcg gcctcatttc ttattctgag ttacaacagt
4441 ccgcaccgct gccggtagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
4501 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc ccaacagtcc
4561 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgccctgca ccattatgtt
4621 ccggatctgc atcgcaggat gctgctggct accctgtgga acacctacat ctgtattaac
4681 gaagcgctaa ccgtttttat catgctctgg gaggcagaat aaatgatcat atcgtcaatt
4741 attacctcca cggggagagc ctgagcaaac tggcctcagg catttgagaa gcacacggtc
4801 acactgcttc cggtagtcaa taaaccggta aaccagcaat agacataagc ggctatttaa
4861 cgaccctgcc ctgaaccgac gaccgggtcg aatttgcttt cgaatttctg ccattcatcc
4921 gcttattatc acttattcag gcgtagcaac caggcgttta agggcaccaa taactgcctt
4981 aaaaaaatta cgccccgccc tgccactcat cgcagttgac tgggtcatgg ctgcgccccg
5041 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
5101 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
5161 gaaacgcgcg aggcagcaga tcaattcgcg cgcgaaggcg aagcggcatg cataatgtgc
5221 ctgtcaaatg gacgaagcag ggattctgca aaccctatgc tactccgtca agccgtcaat
5281 tgtctgattc gttaccaatt atgacaactt gacggctaca tcattcactt tttcttcaca
5341 accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc gcgagaaata
5401 gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc gggtggtgct
5461 caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc
5521 taactgctgg cggaaaagat gtgacagacg cgacggcgac aagcaaacat gctgtgcgac
5581 gctggcgata tcaaaattgc tgtctgccag gtgatcgctg atgtactgac aagcctcgcg
5641 tacccgatta tccatcggtg gatggagcga ctcgttaatc gcttccatgc gccgcagtaa
5701 caattgctca agcagattta tcgccagcag ctccgaatag cgcccttccc cttgcccggc
5761 gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa
5821 gaaccccgta ttggcaaata ttgacggcca gttaagccat tcatgccagt aggcgcgcgg
5881 acgaaagtaa acccactggt gataccattc gcgagcctcc ggatgacgac cgtagtgatg
5941 aatctctcct ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg
6001 atttttcacc accccctgac cgcgaatggt gagattgaga atataacctt tcattcccag
6061 cggtcggtcg ataaaaaaat cgagataacc gttggcctca atcggcgtta aacccgccac
6121 cagatgggca ttaaacgagt atcccggcag caggggatca ttttgcgctt cagccatact
6181 tttcatactc ccgccattca gagaagaaac caattgtcca tattgcatca gacattgccg
6241 tcactgcgtc ttttactggc tcttctcgct aaccaaaccg gtaacccggg accaagttta
6301 ctcatatata cggacagcgg tgcggactgt tgt
//
