| 质粒名称: | pEVOL-pBpF |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 31190 |
| 存储实验室: | Peter Schultz |
|
载体骨架基本信息 |
|
| 载体名称: | p15A |
| 载体抗性: | 氯霉素 |
| 空载体大小: | 5000 bp |
| 载体类型: | 细菌表达 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | M.j. p-benzoylphenylalanine RS (2 copies +tRNA),pEVOL-pBpF |
| 物种来源: | M.jannaschii |
| 插入大小: | 1000 bp |
| 基因突变: | Y32G, E107P, D158T, I159S |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BglII (未破坏) |
| 3' 克隆位点: | SalI (未破坏) |
| 5' 测序引物 | ATTAGCGGATCCTACCTGACGCTTTTTATCGCAACTCTCTACTGTTTCTCCATACCCGTTTTTT |
|
文献引用方法 |
|
| 材料和方法部分: | pEVOL-pBpF was a gift from Peter Schultz (Addgene plasmid # 31190 ; http://n2t.net/addgene:31190 ; RRID:Addgene_31190) |
| 参考文献部分: | Addition of a photocrosslinking amino acid to the genetic code of Escherichiacoli. Chin JW, Martin AB, King DS, Wang L, Schultz PG. Proc Natl Acad Sci U S A. 2002 Aug 20;99(17):11020-4. Epub 2002 Aug 1. 10.1073/pnas.172226299 PubMed 12154230 |
买家导航
pEVOLpBpF载体基本信息
pEVOLpBpF载体图谱质粒图谱和多克隆位点信息
pEVOLpBpF载体简介
pEVOLpBpF载体序列
LOCUS pEVOL-pBpF 6128 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION tRNA synthetase/tRNA pair for the in vivo incorporation of a
photocrosslinker, p-benzoyl-l-phenylalanine into proteins in E coli
in response to the amber codon, TAG. .
ACCESSION .
VERSION .
KEYWORDS pEVOL-pBpF
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6128)
AUTHORS Chin JW, Martin AB, King DS, Wang L, Schultz PG
TITLE Addition of a photocrosslinking amino acid to the genetic code of
Escherichiacoli.
JOURNAL Proc Natl Acad Sci U S A. 2002 Aug 20;99(17):11020-4. Epub 2002 Aug
1.
PUBMED 12154230
REFERENCE 2 (bases 1 to 6128)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6128
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 22..306
/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 229..248
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
CDS 1273..1290
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(1346..1363)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
CDS complement(3266..3925)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 3840..3859
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(3926..4028)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
primer_bind complement(4420..4437)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4554..5099)
/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(4634..4653)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5245..6123)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPSDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
ORIGIN
1 tcatactccc gccattcaga gaagaaacca attgtccata ttgcatcaga cattgccgtc
61 actgcgtctt ttactggctc ttctcgctaa ccaaaccggt aaccccgctt attaaaagca
121 ttctgtaaca aagcgggacc aaagccatga caaaaacgcg taacaaaagt gtctataatc
181 acggcagaaa agtccacatt gattatttgc acggcgtcac actttgctat gccatagcat
241 ttttatccat aagattagcg gatcctacct gacgcttttt atcgcaactc tctactgttt
301 ctccataccc gtttttttgg gctaacagga ggaattagat ctatggacga atttgaaatg
361 ataaagagaa acacatctga aattatcagc gaggaagagt taagagaggt tttaaaaaaa
421 gatgaaaaat ctgctggtat aggttttgaa ccaagtggta aaatacattt agggcattat
481 ctccaaataa aaaagatgat tgatttacaa aatgctggat ttgatataat tatattgttg
541 gctgatttac acgcctattt aaaccagaaa ggagagttgg atgagattag aaaaatagga
601 gattataaca aaaaagtttt tgaagcaatg gggttaaagg caaaatatct ttatggaagt
661 cctttccagc ttgataagga ttatacactg aatgtctata gattggcttt aaaaactacc
721 ttaaaaagag caagaaggag tatggaactt atagcaagag aggatgaaaa tccaaaggtt
781 gctgaagtta tctatccaat aatgcaggtt aatacgagtc attatctggg cgttgatgtt
841 gcagttggag ggatggagca gagaaaaata cacatgttag caagggagct tttaccaaaa
901 aaggttgttt gtattcacaa ccctgtctta acgggtttgg atggagaagg aaagatgagt
961 tcttcaaaag ggaattttat agctgttgat gactctccag aagagattag ggctaagata
1021 aagaaagcat actgcccagc tggagttgtt gaaggaaatc caataatgga gatagctaaa
1081 tacttccttg aatatccttt aaccataaaa aggccagaaa aatttggtgg agatttgaca
1141 gttaatagct atgaggagtt agagagttta tttaaaaata aggaattgca tccaatggat
1201 ttaaaaaatg ctgtagctga agaacttata aagattttag agccaattag aaagagatta
1261 taataagtcg accatcatca tcatcatcat tgagtttaaa cggtctccag cttggctgtt
1321 ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc
1381 tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga
1441 actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag
1501 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc cttgtttgtg
1561 agctcccggt catcaatcat ccccataatc cttgttagat tatcaatttt aaaaaactaa
1621 cagttgtcag cctgtcccgc tttaatatca tacgccgtta tacgttgttt acgctttgag
1681 gaatcccata tggacgaatt tgaaatgata aagagaaaca catctgaaat tatcagcgag
1741 gaagagttaa gagaggtttt aaaaaaagat gaaaaatctg ctggtatagg ttttgaacca
1801 agtggtaaaa tacatttagg gcattatctc caaataaaaa agatgattga tttacaaaat
1861 gctggatttg atataattat attgttggct gatttacacg cctatttaaa ccagaaagga
1921 gagttggatg agattagaaa aataggagat tataacaaaa aagtttttga agcaatgggg
1981 ttaaaggcaa aatatcttta tggaagtcct ttccagcttg ataaggatta tacactgaat
2041 gtctatagat tggctttaaa aactacctta aaaagagcaa gaaggagtat ggaacttata
2101 gcaagagagg atgaaaatcc aaaggttgct gaagttatct atccaataat gcaggttaat
2161 acgagtcatt atctgggcgt tgatgttgca gttggaggga tggagcagag aaaaatacac
2221 atgttagcaa gggagctttt accaaaaaag gttgtttgta ttcacaaccc tgtcttaacg
2281 ggtttggatg gagaaggaaa gatgagttct tcaaaaggga attttatagc tgttgatgac
2341 tctccagaag agattagggc taagataaag aaagcatact gcccagctgg agttgttgaa
2401 ggaaatccaa taatggagat agctaaatac ttccttgaat atcctttaac cataaaaagg
2461 ccagaaaaat ttggtggaga tttgacagtt aatagctatg aggagttaga gagtttattt
2521 aaaaataagg aattgcatcc aatggattta aaaaatgctg tagctgaaga acttataaag
2581 attttagagc caattagaaa gagattataa ctgcagtttc aaacgctaaa ttgcctgatg
2641 cgctacgctt atcaggccta catgatctct gcaatatatt gagtttgcgt gcttttgtag
2701 gccggataag gcgttcacgc cgcatccggc aagaaacagc aaacaatcca aaacgccgcg
2761 ttcagcggcg ttttttctgc ttttcttcgc gaattaattc cgcttcgcaa catgtgagca
2821 ccggtttatt gactaccgga agcagtgtga ccgtgtgctt ctcaaatgcc tgaggccagt
2881 ttgctcaggc tctccccgtg gaggtaataa ttgacgatat gatcagtgca cggctaacta
2941 agcggcctgc tgactttctc gccgatcaaa aggcattttg ctattaaggg attgacgagg
3001 gcgtatctgc gcagtaagat gcgccccgca ttccggcggt agttcagcag ggcagaacgg
3061 cggactctaa atccgcatgg caggggttca aatcccctcc gccggaccaa attcgaaaag
3121 cctgctcaac gagcaggctt ttttgcatgc tcgagcagct cagggtcgaa tttgctttcg
3181 aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaacca ggcgtttaag
3241 ggcaccaata actgccttaa aaaaattacg ccccgccctg ccactcatcg cagtactgtt
3301 gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa
3361 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg
3421 gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg
3481 gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt
3541 caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt
3601 attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt
3661 gaacactatc ccatatcacc agctcaccgt ctttcattgc catacggaat tccggatgag
3721 cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct
3781 ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag
3841 caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg
3901 tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact
3961 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
4021 gccgatcaac gtctcatttt cgccaaaagt tggcccaggg cttcccggta tcaacaggga
4081 caccaggatt tatttattct gcgaagtgat cttccgtcac aggtatttat tcggcgcaaa
4141 gtgcgtcggg tgatgctgcc aacttactga tttagtgtat gatggtgttt ttgaggtgct
4201 ccagtggctt ctgtttctat cagctgtccc tcctgttcag ctactgacgg ggtggtgcgt
4261 aacggcaaaa gcaccgccgg acatcagcgc tagcggagtg tatactggct tactatgttg
4321 gcactgatga gggtgtcagt gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt
4381 gcgtcagcag aatatgtgat acaggatata ttccgcttcc tcgctcactg actcgctacg
4441 ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa cggggcggag atttcctgga
4501 agatgccagg aagatactta acagggaagt gagagggccg cggcaaagcc gtttttccat
4561 aggctccgcc cccctgacaa gcatcacgaa atctgacgct caaatcagtg gtggcgaaac
4621 ccgacaggac tataaagata ccaggcgttt ccccctggcg gctccctcgt gcgctctcct
4681 gttcctgcct ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc
4741 acgcctgaca ctcagttccg ggtaggcagt tcgctccaag ctggactgta tgcacgaacc
4801 ccccgttcag tccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccgga
4861 aagacatgca aaagcaccac tggcagcagc cactggtaat tgatttagag gagttagtct
4921 tgaagtcatg cgccggttaa ggctaaactg aaaggacaag ttttggtgac tgcgctcctc
4981 caagccagtt acctcggttc aaagagttgg tagctcagag aaccttcgaa aaaccgccct
5041 gcaaggcggt tttttcgttt tcagagcaag agattacgcg cagaccaaaa cgatctcaag
5101 aagatcatct tattaatcag ataaaatatt tctagatttc agtgcaattt atctcttcaa
5161 atgtagcacc tgaagtcagc cccatacgat ataagttgta attctcatgt ttgacagctt
5221 atcatcgata agcttggtac ccaattatga caacttgacg gctacatcat tcactttttc
5281 ttcacaaccg gcacggaact cgctcgggct ggccccggtg cattttttaa atacccgcga
5341 gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag gcatccgggt
5401 ggtgctcaaa agcagcttcg cctggctgat acgttggtcc tcgcgccagc ttaagacgct
5461 aatccctaac tgctggcgga aaagatgtga cagacgcgac ggcgacaagc aaacatgctg
5521 tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt actgacaagc
5581 ctcgcgtacc cgattatcca tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg
5641 cagtaacaat tgctcaagca gatttatcgc cagcagctcc gaatagcgcc cttccccttg
5701 cccggcgtta atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg
5761 gcgaaagaac cccgtattgg caaatattga cggccagtta agccattcat gccagtaggc
5821 gcgcggacga aagtaaaccc actggtgata ccattcgcga gcctccggat gacgaccgta
5881 gtgatgaatc tctcctggcg ggaacagcaa aatatcactc ggtcggcaaa caaattctcg
5941 tccctgattt ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat aacctttcat
6001 tcccagcggt cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc
6061 cgccaccaga tgggcattaa acgagtatcc cggcagcagg ggatcatttt gcgcttcagc
6121 catacttt
//
