| 质粒名称: | pJBEI-6411 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 47050 |
| 存储实验室: | Taek Soon Lee |
|
载体骨架基本信息 |
|
| 载体名称: | pBbB8k |
| 载体出品公司: | -- |
| 空载体大小: | 2300 bp |
| 完整质粒大小: | 6656 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 Synthetic Biology ; BglBrick |
| 筛选标记: | -- |
| 载体抗性: | 卡那霉素，50μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | 编码细胞色素P450（CYP153A6）的ahpG-ahpH-ahpI及其相关的铁氧还蛋白和铁氧还素还原酶 |
| 物种来源: | 鞘氨醇 |
| 插入大小: | 4358 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | Pbad |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoRI, BglII (未破坏) |
| 3’克隆位点: | BamHI, XhoI(未破坏) |
| 5’测序引物 | F8 |
| 3’测序引物 | R1 |
|
文献引用方法 |
|
| 材料和方法部分: | pJBEI-6411 was a gift from Taek Soon Lee (Addgene plasmid # 47050 ; http://n2t.net/addgene:47050 ; RRID:Addgene_47050) |
| 参考文献部分: | Metabolic engineering of Escherichia coli for limonene and perillyl alcohol production. Alonso-Gutierrez J, Chan R, Batth TS, Adams PD, Keasling JD, Petzold CJ, Lee TS. Metab Eng. 2013 May 29;19C:33-41. doi: 10.1016/j.ymben.2013.05.004. 10.1016/j.ymben.2013.05.004 PubMed 23727191 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pJBEI-6411 6656 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION BglBrick plasmid (=pBbB8k-P450) coding for a cytochrome P450 and its
associated reductases to oxidize limonene to perillyl alcohol in E.
coli.
ACCESSION .
VERSION .
KEYWORDS pJBEI-6411
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6656)
AUTHORS Alonso-Gutierrez J, Chan R, Batth TS, Adams PD, Keasling JD, Petzold
CJ, Lee TS
TITLE Metabolic engineering of Escherichia coli for limonene and perillyl
alcohol production.
JOURNAL Metab Eng. 2013 May 29;19C:33-41. doi: 10.1016/j.ymben.2013.05.004.
PUBMED 23727191
REFERENCE 2 (bases 1 to 6656)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6656
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(7..885)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
promoter 912..1196
/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 1119..1138
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
terminator 4171..4242
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 4258..4285
/label=T7Te terminator
/note="phage T7 early transcription terminator"
CDS complement(4352..5014)
/codon_start=1
/product="replication protein for the broad-host-range
plasmid pBBR1 from Bordetella bronchiseptica"
/label=pBBR1 Rep
/translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVLAYVV
NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
terminator 5601..5695
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(5726..6520)
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(5837..5856)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 6447..6466
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
ORIGIN
1 gacgtcttat gacaacttga cggctacatc attcactttt tcttcacaac cggcacggaa
61 ctcgctcggg ctggccccgg tgcatttttt aaatacccgc gagaaataga gttgatcgtc
121 aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca aaagcagctt
181 cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta actgctggcg
241 gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc
301 aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta cccgattatc
361 catcggtgga tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag
421 cagatttatc gccagcagct ccgaatagcg cccttcccct tgcccggcgt taatgatttg
481 cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga accccgtatt
541 ggcaaatatt gacggccagt taagccattc atgccagtag gcgcgcggac gaaagtaaac
601 ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa tctctcctgg
661 cgggaacagc aaaatatcac ccggtcggca aacaaattct cgtccctgat ttttcaccac
721 cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat
781 aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt
841 aaacgagtat cccggcagca ggggatcatt ttgcgcttca gccatacttt tcatactccc
901 gccattcaga gaagaaacca attgtccata ttgcatcaga cattgccgtc actgcgtctt
961 ttactggctc ttctcgctaa ccaaaccggt aaccccgctt attaaaagca ttctgtaaca
1021 aagcgggacc aaagccatga caaaaacgcg taacaaaagt gtctataatc acggcagaaa
1081 agtccacatt gattatttgc acggcgtcac actttgctat gccatagcat ttttatccat
1141 aagattagcg gattctacct gacgcttttt atcgcaactc tctactgttt ctccataccc
1201 gtttttttgg gaattcaaaa gatctaggag gataaagaaa tgactgaaat gactgtcgcg
1261 gcttcggacg caactaatgc cgcctacggc atggccctgg aagatattga tgttagcaat
1321 ccggttctgt tccgcgataa tacgtggcac ccctatttca aaaggctgcg tgaggaagac
1381 ccagtgcact attgtaagtc ctctatgttt ggtccatact ggagtgttac caaatatcgt
1441 gatataatgg ccgtggaaac gaacccgaaa gtattttcaa gcgaggctaa atcaggtggt
1501 attaccataa tggatgataa cgccgctgct agcctgccta tgtttatcgc gatggacccc
1561 ccgaaacatg atgttcagcg aaaaaccgta tcccctattg ttgcgccgga gaatttagcc
1621 acgatggagt cagtcatacg ccagagaacg gctgacttgc ttgatggttt acccataaat
1681 gaagaatttg actgggttca tcgcgtctcc attgagttaa ccaccaaaat gctggcgacc
1741 ctgtttgact tcccctggga cgatagagcg aaattgactc gctggtccga tgtgaccaca
1801 gctttacccg gaggtggtat aattgatagc gaagaacagc gtatggccga gttgatggag
1861 tgcgctacgt acttcaccga actgtggaat cagcgtgtta acgctgagcc aaaaaatgat
1921 ctgatctcta tgatggcaca ctcggagagt actagacata tggctccgga ggaatactta
1981 gggaacatcg tgctgctcat cgtaggaggg aatgatacga cccgtaactc catgacgggt
2041 ggtgtacttg ccctaaacga atttcccgat gaatatcgga agctaagtgc gaatcctgca
2101 ttgatctcgt caatggtgtc cgaaatcatt cggtggcaga cccctctctc ccatatgcgc
2161 cggacggctc ttgaagacat agagtttggt ggaaaacata ttagacaagg agacaaggtg
2221 gttatgtggt atgtttccgg caatcgagat ccagaagcta tcgacaaccc tgatacattt
2281 attatcgatc gtgcaaagcc acgtcaacac ctgtcttttg gcttcggtat tcatcgttgc
2341 gttggtaatc gcctggcgga attacaatta aatattctgt gggaagagat tctgaaacga
2401 tggccagacc cccttcagat tcaagtttta caggaaccaa cgcgtgttct gagtcctttt
2461 gtaaaaggtt atgaatcgct gccagtacgc atcaacgcgt aaggatctag gaggataaag
2521 aaatgataca tacaggcgtc acggaggctg ttgtggtcgt cggtgctggg caggcgggag
2581 cacagactgt gacaagtttg cgacagcgcg gttttgaagg tcagattacg ctgttaggag
2641 atgaacccgc gttaccgtat cagcgcccac cgttatccaa agcatttttg gctggcacac
2701 tgccactgga tcgcctgtat ttacgcccag ccgcgtttta ccaacaggca catgtggatg
2761 tcatggtcga tacaggtgtc tcggaactag ataccgaaaa tcgtcgtatc cggctgacag
2821 atggacgagc gatatctttt gaccatctgg ttttagccac cggggggagg cccagacctt
2881 tagcctgtcc cggagcggac cacccccgcg ttcattattt gcgtacagtc accgatgttg
2941 atcgtattcg gtcacagttt catcctggca cccgacttgt attagtgggg ggggggtaca
3001 tagggttaga aattgcagcg gtagccgccg aattaggttt gactgtaaca gtgcttgagg
3061 cccagacaac tgtgttggcc cgcgttacgt gtccaaccgt cgcacgcttt ttcgagcata
3121 cacaccggcg cgcgggtgtc accattcgct gcgccacgac cgtgacacgg attcatgatt
3181 cttcgagtac tgctcgaatc gagctcgact caggggaata tattgacgcg gatctggtta
3241 tagtgggtat tggtttgctc cccaacgttg accttgcaag cgcagctggc ctcacgtgcg
3301 aaagtggcat tgtggtagac agtcgttgcc aaacgtccgc acctggaatc tacgcagccg
3361 gcgattgcac gcaatatcca tcccctatct acggtcggcc acttcatttg gaatctgttc
3421 ataatgcgat cgaacaggct aagaccgcag cagccgcaat cttggggcgg gatgaaccgt
3481 ttcgtcaggt tccctggttc tggtcggatc agtacaacat taaacttcag accgcaggcg
3541 ttaatgaggg ttacgatgat gtcatcattc gcggtgaccc cgcatcagcg agctttgccg
3601 cattttactt aagagcgggg aaattactgg ctgtggatgc gattaatcgg ccgcgtgagt
3661 tcatggcttc caaaaccctg attgctgaaa gggccgaagt agacccgacc caattggcag
3721 acgagagctt gcctccgacc gccctggcgg cagcggtcaa tggtcctact cgggctacga
3781 gtcccacaag cttataagga tctaggagga taaagaaatg cccaagatta cgtacattga
3841 ttacacggga acgtcacgct gtgttgatgc tgaaaatggt atgtcattaa tggaaattgc
3901 gataaacaat aacgttcccg gaattgatgg agattgcggc ggcgagtgcg cgtgtgccac
3961 gtgtcatgtg catgttgacg ccgattggct ggataaatta cctccttcca gcgaccagga
4021 ggttagtatg ttggaatttt gtgacggtgt tgatcatact tcccgcttag ggtgtcagat
4081 taaaatttgc ccgaccttag atggtattgt ggtacggacg cccgccgcgc agcactaagg
4141 atccaaactc gagtaaggat ctccaggcat caaataaaac gaaaggctca gtcgaaagac
4201 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag tcacactggc
4261 tcaccttcgg gtgggccttt ctgcgtttat acctaggcta cagccgatag tctggaacag
4321 cgcacttacg ggttgctgcg caacccaagt gctaccggcg cggcagcgtg acccgtgtcg
4381 gcggctccaa cggctcgcca tcgtccagaa aacacggctc atcgggcatc ggcaggcgct
4441 gctgcccgcg ccgttcccat tcctccgttt cggtcaaggc tggcaggtct ggttccatgc
4501 ccggaatgcc gggctggctg ggcggctcct cgccggggcc ggtcggtagt tgctgctcgc
4561 ccggatacag ggtcgggatg cggcgcaggt cgccatgccc caacagcgat tcgtcctggt
4621 cgtcgtgatc aaccaccacg gcggcactga acaccgacag gcgcaactgg tcgcggggct
4681 ggccccacgc cacgcggtca ttgaccacgt aggccaacac ggtgccgggg ccgttgagct
4741 tcacgacgga gatccagcgc tcggccacca agtccttgac tgcgtattgg accgtccgca
4801 aagaacgtcc gatgagcttg gaaagtgtct tctggctgac caccacggcg ttctggtggc
4861 ccatctgcgc cacgaggtga tgcagcagca ttgccgccgt gggtttcctc gcaataagcc
4921 cggcccacgc ctcatgcgct ttgcgttccg tttgcaccca gtgaccgggc ttgttcttgg
4981 cttgaatgcc gatttctctg gactgcgtgg ccatgcttat ctccatgcgg taggggtgcc
5041 gcacggttgc ggcaccatgc gcaatcagct gcaacttttc ggcagcgcga caacaattat
5101 gcgttgcgta aaagtggcag tcaattacag attttcttta acctacgcaa tgagctattg
5161 cggggggtgc cgcaatgagc tgttgcgtac cccccttttt taagttgttg atttttaagt
5221 ctttcgcatt tcgccctata tctagttctt tggtgcccaa agaagggcac ccctgcgggg
5281 ttcccccacg ccttcggcgc ggctccccct ccggcaaaaa gtggcccctc cggggcttgt
5341 tgatcgactg cgcggccttc ggccttgccc aaggtggcgc tgcccccttg gaacccccgc
5401 actcgccgcc gtgaggctcg gggggcaggc gggcgggctt cgcccttcga ctgcccccac
5461 tcgcataggc ttgggtcgtt ccaggcgcgt caaggccaag ccgctgcgcg gtcgctgcgc
5521 gagccttgac ccgccttcca cttggtgtcc aaccggcaag cgaagcgcgc aggccgcagg
5581 ccggaggcac tagtgcttgg attctcacca ataaaaaacg cccggcggca accgagcgtt
5641 ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag gagtccaagc
5701 gagctctcga accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga
5761 tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc
5821 cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca
5881 cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg
5941 gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga
6001 gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat
6061 cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt
6121 cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg
6181 atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca
6241 atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc
6301 ccgtcgtggc cagccacgat agccgcgctg cctcgtcctg cagttcattc agggcaccgg
6361 acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg
6421 catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag
6481 cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg
6541 tctcttgatc agatcatgat cccctgcgcc atcagatcct tggcggcaag aaagccatcc
6601 agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc aattcc
//
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