| 质粒名称: | pDule2-pCNF |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 85495 |
| 存储实验室: | Ryan Mehl |
|
载体骨架基本信息 |
|
| 载体名称: | pDule2 |
| 载体出品公司: | -- |
| 载体抗性: | 大观霉素 |
| 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: | pDule2-pCNF was a gift from Ryan Mehl (Addgene plasmid # 85495 ; http://n2t.net/addgene:85495 ; RRID:Addgene_85495) |
| 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
pDule2pCNF载体基本信息
pDule2pCNF载体图谱质粒图谱和多克隆位点信息
pDule2pCNF载体简介
pDule2pCNF载体序列
LOCUS pDule2-pCNF 7003 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 pDule2-pCNF
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7003)
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 7003)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7003
/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 2572..2663
/gene="bla"
/label=AmpR promoter
CDS 2664..3455
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
primer_bind complement(3631..3650)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 4776..4798
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 4790..4807
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 4791..4807
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 5791..5813
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(5969..6847)
/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 tgcgaccgag tgagctagct atttgtttat
2581 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
2641 aataatattg aaaaaggaag agtatgaggg aagcggtgat cgccgaagta tcgactcaac
2701 tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt
2761 tgtacggctc cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta
2821 cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa
2881 cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc
2941 acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc
3001 agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta
3061 tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac
3121 tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat
3181 ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca
3241 tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa
3301 tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg
3361 gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg
3421 tgaaaggcga gatcaccaag gtagtcggca aataatgtct aacaattcgt tcaagccgag
3481 gggccgcaag atccggccac gatgacccgg tcgtcggttc agggcagggt cgttaaatag
3541 ccgcttatgt ctattgctgg tttaccggga tcctctacgc cggacgcatc gtggccggca
3601 tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag
3661 atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc
3721 ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg
3781 tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag
3841 agcgtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg
3901 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg
3961 tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga
4021 tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg
4081 gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccgacg
4141 cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc cccattatga
4201 ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg
4261 tagatgacga ccatcaggga cagcttcaag gatcgctcgc ggctcttacc agcctaactt
4321 cgatcattgg accgctgatc gtcacggcga tttatgccgc ctcggcgagc acatggaacg
4381 ggttggcatg gattgtaggc gccgccctat accttgtctg cctccccgcg ttgcgtcgcg
4441 gtgcatggag ccgggccacc tcgacctgaa tggaagccgg cggcacctcg ctaacggatt
4501 caccactcca agaattggag ccaatcaatt cttgcggaga actgtgaatg cgcaaaccaa
4561 cccttggcag aacatatcca tcgcgtccgc catctccagc agccgcacgc ggcgcatctc
4621 gggctccttg catgcaccat tccttgcggc ggcggtgctc aacggcctca acctactact
4681 gggctgcttc ctaatgcagg agtcgcataa gggagagcgt ctggcgaaag ggggatgtgc
4741 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
4801 ggccagtgcc aagcttaaaa aaaatcctta gctttcgcta aggatctgca gtggtccggc
4861 gggccggatt tgaaccagcg ccatgcggat ttagagtccg ccgttctgcc ctgctgaact
4921 accgccggga attcagcgtt acaagtatta cacaaagttt tttatgttga gaatattttt
4981 ttgatggggc gccacttatt tttgatcgtt cgctcaaaga agcggcgcca gggttgtttt
5041 tcttttcacc agtgagacgg gcaacagaac gccatgagcg gcctcatttc ttattctgag
5101 ttacaacagt ccgcaccgct gccggtagct ccttccggtg ggcgcggggc atgactatcg
5161 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc
5221 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgccctgca
5281 ccattatgtt ccggatctgc atcgcaggat gctgctggct accctgtgga acacctacat
5341 ctgtattaac gaagcgctaa ccgtttttat catgctctgg gaggcagaat aaatgatcat
5401 atcgtcaatt attacctcca cggggagagc ctgagcaaac tggcctcagg catttgagaa
5461 gcacacggtc acactgcttc cggtagtcaa taaaccggta aaccagcaat agacataagc
5521 ggctatttaa cgaccctgcc ctgaaccgac gaccgggtcg aatttgcttt cgaatttctg
5581 ccattcatcc gcttattatc acttattcag gcgtagcaac caggcgttta agggcaccaa
5641 taactgcctt aaaaaaatta cgccccgccc tgccactcat cgcagttgac tgggtcatgg
5701 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
5761 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
5821 cgtcatcacc gaaacgcgcg aggcagcaga tcaattcgcg cgcgaaggcg aagcggcatg
5881 cataatgtgc ctgtcaaatg gacgaagcag ggattctgca aaccctatgc tactccgtca
5941 agccgtcaat tgtctgattc gttaccaatt atgacaactt gacggctaca tcattcactt
6001 tttcttcaca accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc
6061 gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc
6121 gggtggtgct caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga
6181 cgctaatccc taactgctgg cggaaaagat gtgacagacg cgacggcgac aagcaaacat
6241 gctgtgcgac gctggcgata tcaaaattgc tgtctgccag gtgatcgctg atgtactgac
6301 aagcctcgcg tacccgatta tccatcggtg gatggagcga ctcgttaatc gcttccatgc
6361 gccgcagtaa caattgctca agcagattta tcgccagcag ctccgaatag cgcccttccc
6421 cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat
6481 ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca gttaagccat tcatgccagt
6541 aggcgcgcgg acgaaagtaa acccactggt gataccattc gcgagcctcc ggatgacgac
6601 cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt
6661 ctcgtccctg atttttcacc accccctgac cgcgaatggt gagattgaga atataacctt
6721 tcattcccag cggtcggtcg ataaaaaaat cgagataacc gttggcctca atcggcgtta
6781 aacccgccac cagatgggca ttaaacgagt atcccggcag caggggatca ttttgcgctt
6841 cagccatact tttcatactc ccgccattca gagaagaaac caattgtcca tattgcatca
6901 gacattgccg tcactgcgtc ttttactggc tcttctcgct aaccaaaccg gtaacccggg
6961 accaagttta ctcatatata cggacagcgg tgcggactgt tgt
//
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