| 质粒名称: | pSP6-RNAP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 48143 |
| 存储实验室: | Dieter Jahn |
|
载体骨架基本信息 |
|
| 载体名称: | pMGBm19 |
| 载体出品公司: | Gamer et al., 2009 |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达，合成生物学 |
| 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: | rna polymerase SP6 |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 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 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pSP6-RNAP was a gift from Dieter Jahn (Addgene plasmid # 48143 ; http://n2t.net/addgene:48143 ; RRID:Addgene_48143) |
| 参考文献部分: | Application of Escherichia coli phage K1E DNA-dependent RNA polymerase for in vitro RNA synthesis and in vivo protein production in Bacillus megaterium. Stammen S, Schuller F, Dietrich S, Gamer M, Biedendieck R, Jahn D. Appl Microbiol Biotechnol. 2010 Sep;88(2):529-39. doi: 10.1007/s00253-010-2732-y. Epub 2010 Jul 2. 10.1007/s00253-010-2732-y PubMed 20596705 |
买家导航
pSP6RNAP载体基本信息
pSP6RNAP载体图谱质粒图谱和多克隆位点信息
pSP6RNAP载体简介
pSP6RNAP载体序列
LOCUS pSP6-RNAP 8784 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Shuttle vector E. coli/B.meg.; encoding the RNA polymerase of the
phage SP6 under control of the xylose inducible promoter PxylA.
ACCESSION .
VERSION .
KEYWORDS pSP6-RNAP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8784)
AUTHORS Stammen S, Schuller F, Dietrich S, Gamer M, Biedendieck R, Jahn D
TITLE Application of Escherichia coli phage K1E DNA-dependent RNA
polymerase for in vitro RNA synthesis and in vivo protein production
in Bacillus megaterium.
JOURNAL Appl Microbiol Biotechnol. 2010 Sep;88(2):529-39. doi:
10.1007/s00253-010-2732-y. Epub 2010 Jul 2.
PUBMED 20596705
REFERENCE 2 (bases 1 to 8784)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8784
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(3804..3820)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3804..3820)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3817..3839)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3828..3844
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(3852..3882)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3897..3918
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4035..4052)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4206..4794)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4286..4305)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4965..5825)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 5588..5607
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5826..5930)
/gene="bla"
/label=AmpR promoter
primer_bind 5998..6016
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(6054..6076)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 6176..6195
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 caagatttac acgctatcca gcttcaatta gaagaagaga tgtttaatgg tggcattcgt
61 cgcttcgaag cagatcaaca acgccagatt gcagcaggta gcgagagcga cacagcatgg
121 aaccgccgcc tgttgtcaga acttattgca cctatggctg aaggcattca ggcttataaa
181 gaagagtacg aaggtaagaa aggtcgtgca cctcgcgcat tggctttctt acaatgtgta
241 gaaaatgaag ttgcagcata catcactatg aaagttgtta tggatatgct gaatacggat
301 gctacccttc aggctattgc aatgagtgta gcagaacgca ttgaagacca agtgcgcttt
361 tctaagctag aaggtcacgc cgctaaatac tttgagaagg ttaagaagtc actcaaggct
421 agccgtacta agtcatatcg tcacgctcat aacgtagctg tagttgctga aaaatcagtt
481 gcagaaaagg acgcggactt tgaccgttgg gaggcgtggc caaaagaaac tcaattgcag
541 attggtacta ccttgcttga aatcttagaa ggtagcgttt tctataatgg tgaacctgta
601 tttatgcgtg ctatgcgcac ttatggcgga aagactattt actacttaca aacttctgaa
661 agtgtaggcc agtggattag cgcattcaaa gagcacgtag cgcaattaag cccagcttat
721 gccccttgcg taatccctcc tcgtccttgg agaactccat ttaatggagg gttccatact
781 gagaaggtag ctagccgtat ccgtcttgta aaaggtaacc gtgagcatgt acgcaagttg
841 actcaaaagc aaatgccaaa ggtttataag gctatcaacg cattacaaaa tacacaatgg
901 caaatcaaca aggatgtatt agcagttatt gaagaagtaa tccgcttaga ccttggttat
961 ggtgtacctt ccttcaagcc actgattgac aaggagaaca agccagctaa cccggtacct
1021 gttgaattcc aacacctgcg cggtcgtgaa ctgaaagaga tgctatcacc tgagcagtgg
1081 caacaattca ttaactggaa aggcgaatgc gcgcgcctat ataccgcaga aactaagcgc
1141 ggttcaaagt ccgccgccgt tgttcgcatg gtaggacagg cccgtaaata tagcgccttt
1201 gaatccattt acttcgtgta cgcaatggat agccgcagcc gtgtctatgt gcaatctagc
1261 acgctctctc cgcagtctaa cgacttaggt aaggcattac tccgctttac cgagggacgc
1321 cctgtgaatg gcgtagaagc gcttaaatgg ttctgcatca atggtgctaa cctttgggga
1381 tgggacaaga aaacttttga tgtgcgcgtg tctaacgtat tagatgagga attccaagat
1441 atgtgtcgag acatcgccgc agaccctctc acattcaccc aatgggctaa agctgatgca
1501 ccttatgaat tcctcgcttg gtgctttgag tatgctcaat accttgattt ggtggatgaa
1561 ggaagggccg acgaattccg cactcaccta ccagtacatc aggacgggtc ttgttcaggc
1621 attcagcact atagtgctat gcttcgcgac gaagtagggg ccaaagctgt taacctgaaa
1681 ccctccgatg caccgcagga tatctatggg gcggtggcgc aagtggttat caagaagaat
1741 gcgctatata tggatgcgga cgatgcaacc acgtttactt ctggtagcgt cacgctgtcc
1801 ggtacagaac tgcgagcaat ggctagcgca tgggatagta ttggtattac ccgtagctta
1861 accaaaaagc ccgtgatgac cttgccatat ggttctactc gcttaacttg ccgtgaatct
1921 gtgattgatt acatcgtaga cttagaggaa aaagaggcgc agaaggcagt agcagaaggg
1981 cggacggcaa acaaggtaca tccttttgaa gacgatcgtc aagattactt gactccgggc
2041 gcagcttaca actacatgac ggcactaatc tggccttcta tttctgaagt agttaaggca
2101 ccgatagtag ctatgaagat gatacgccag cttgcacgct ttgcagcgaa acgtaatgaa
2161 ggcctgatgt acaccctgcc tactggcttc atcttagaac agaagatcat ggcaaccgag
2221 atgctacgcg tgcgtacctg tctgatgggt gatatcaaga tgtcccttca ggttgaaacg
2281 gatatcgtag atgaagccgc tatgatggga gcagcagcac ctaatttcgt acacggtcat
2341 gacgcaagtc accttatcct taccgtatgt gaattggtag acaagggcgt aactagtatc
2401 gctgtaatcc acgactcttt tggtactcat gcagacaaca ccctcactct tagagtggca
2461 cttaaagggc agatggttgc aatgtatatt gatggtaatg cgcttcagaa actactggag
2521 gagcatgaag agcgctggat ggttgataca ggtatcgaag tacctgagca aggggagttc
2581 gaccttaacg aaatcatgga ttctgaatac gtatttgcct aaggatcctc tagagtcatt
2641 tatattcaac tgttaactaa aactgtctgc aaattaatca atgtctctag taaaattccg
2701 attttatacc cactcctatt ttcatcaata ggacatgata taataaaaaa gaacggagat
2761 tgaggcctcc gttctcttgg ttgtacttgt tgcaatagta cgtttatttg gttacgtata
2821 ttatagcaag tactccaaca aaaaatcaat cgttggagga atttagaaat gaatggaatt
2881 atcaaaacaa taaaaacaac gcaatatgct caaatacata acgcgccatt acaaacggac
2941 ctggaggact tgcggtctat cggtctatta agtcatatta tgagcttagg tgaaggttgg
3001 acgattagga agacccagtt acaaaacaaa ttttctcgta gaaatgtaga cgctgcttgg
3061 aaggaattag ctattaaaca atacgctgtc ggttttagcg catatgttga cggaaaaaaa
3121 gattattttt acgccgtatc agatattccg atttcgcaag ctaatttcga attgcttgtc
3181 attgaacaaa taaatctctt acaatcgcaa ggtaaaaacg tcatgacgtt aagcgtgata
3241 caacattcgc cacttgaaat cactgaaaaa atatctgatg tacaaaatgt acatcagata
3301 caaaataagc aaatttcttc tgatgtacgt tctgtacaac acagtgtgta caacacagaa
3361 cgtacaccta taaataaaaa agaaacaaat gaaaaattaa caaatgaaaa aattagtagt
3421 agtccacgtg attttattga tgataaatta cgagaaaaat ataataatgt acctttcgac
3481 gaagtgaaaa gtgaaatgct taacgatact gtaatcgttg atacgaataa gcaatataaa
3541 tcactattag aataccgtct taaacattgg aaaccgaagc aaacaaaaag gaaacgtgta
3601 gtacgtaatg cacgtaaaga aatggttcca aagtggctac aaacaaatga tgaaaatgca
3661 gtagcagaag atcttccagt gaataatgat tttgaagctg aaaaggaaaa aatgaaggcg
3721 gagcttatgc aattagatgc agaattaaaa gcaggtaatc gttaataaat gacctgcagg
3781 catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
3841 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
3901 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
3961 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
4021 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
4081 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
4141 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
4201 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
4261 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
4321 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
4381 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
4441 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
4501 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
4561 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
4621 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
4681 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
4741 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
4801 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
4861 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
4921 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
4981 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
5041 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
5101 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
5161 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
5221 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
5281 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
5341 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
5401 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
5461 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
5521 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
5581 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
5641 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
5701 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
5761 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
5821 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
5881 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
5941 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
6001 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
6061 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
6121 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
6181 tcagagcaga ttgtactgag agtgcaccat agatctggag ctgtaatata aaaaccttct
6241 tcaactaacg gggcaggtta gtgacattag aaaaccgact gtaaaaagta cagtcggcat
6301 tatctcatat tataaaagcc agtcattagg cctatctgac aattcctgaa tagagttcat
6361 aaacaatcct gcatgataac catcacaaac agaatgatgt acctgtaaag atagcggtaa
6421 atatattgaa ttacctttat taatgaattt tcctgctgta ataatgggta gaaggtaatt
6481 actattatta ttgatattta agttaaaccc agtaaatgaa gtccatggaa taatagaaag
6541 agaaaaagca ttttcaggta taggtgtttt gggaaacaat ttccccgaac cattatattt
6601 ctctacatca gaaaggtata aatcataaaa ctctttgaag tcattcttta caggagtcca
6661 aataccagag aatgttttag atacaccatc aaaaattgta taaagtggct ctaacttatc
6721 ccaataacct aactctccgt cgctattgta accagttcta aaagctgtat ttgagtttat
6781 cacccttgtc actaagaaaa taaatgcagg gtaaaattta tatccttctt gttttatgtt
6841 tcggtataaa acactaatat caatttctgt ggttatacta aaagtcgttt gttggttcaa
6901 ataatgatta aatatctctt ttctcttcca attgtctaaa tcaattttat taaagttcat
6961 ttgatatgcc tcctaaattt ttatctaaag tgaatttagg aggcttactt gtctgctttc
7021 ttcattagaa tcaatccttt tttaaaagtc aatattactg taacataaat atatatttta
7081 aaaatatccc actttatcca attttcgttt gttgaactaa tgggtgcttt agttgaagaa
7141 taaaagacca cattaaaaaa tgtggtcttt tgtgtttttt taaaggattt gagcgtagcg
7201 aaaaatcctt ttctttctta tcttgataat aagggtaact attgccggat cgtcctcagg
7261 agtaggcgac atcgctaaat aatgatctat gcggtgtgaa ataccgcaca gatgcgtaag
7321 gagaaaatac cgcatcaggc gccctttgcg ttcacttaac taacttatag gggtaacact
7381 taaaaaagaa tcaataacga tagaaaccgc tcctaaagca ggtgcatttt ttcctaacga
7441 agaaggcaat agttcacatt tattgtctaa atgagaatgg actctagaag aaacttcgtt
7501 tttaatcgta tttaaaacaa tgggatgaga ttcaattata tgatttctca agataacagc
7561 ttctatatca aatgtattaa ggatattggt taatccaatt ccgatataaa agccaaagtt
7621 ttgaagtgca tttaacattt ctacatcatt tttatttgcg cgttccacaa tctcttttcg
7681 agaaatattc ttttcttctt tagagagcga agccagtaac gctttttcag aagcatataa
7741 ttcccaacag cctcgatttc cacagctgca tttgggtcca ttaaaatcta tcgtcatatg
7801 acccatttcc ccagaaaaac cctgaacacc tttatacaat tcgttgttaa taacaagtcc
7861 agttccaatt ccgatattaa tactgatgta aacgatgttt tcatagtttt ttgtcatacc
7921 aaatactttt tcaccgtatg ctcctgcatt agcttcattt tcaacaaaaa ccggaacatt
7981 aaactcactc tcaattaaaa actgcaaatc tttgatattc caatttaagt taggcatgaa
8041 aataatttgc tgatgacgat ctacaaggcc tggaacacaa attcctattc cgactagacc
8101 ataaggggac tcaggcatat gggttacaaa accatgaata agtgcaaata aaatctcttt
8161 tacttcacta gcggaagaac tagacaagtc agaagtcttc tcgagaataa tatttccttc
8221 taagtcggtt agaattccgt taagatagtc gactcctata tcaataccaa tcgagtagcc
8281 tgcattctta ttaaaaacaa gcattacagg tcttctgccg cctctagatt gccctgcccc
8341 aatttcaaaa ataaaatctt tttcaagcag tgtatttact tgagaggaga cagtagactt
8401 gtttaatcct gtaatctcag agagagttgc cctggagaca ggggagttct tcaaaatttc
8461 atctaatatt aatttttgat tcattttttt tactaaagct tgatctgcaa tttgaataat
8521 aaccactcct ttgtttatcc accgaactaa gttggtgttt tttgaagctt gaattagata
8581 tttaaaagta tcatatctaa tattataact aaattttcta aaaaaaacat tgaaataaac
8641 atttattttg tatatgatga gataaagtta gtttattgga taaacaaact aactcaatta
8701 agatagttga tggataaact tgttcactta aatcaaaggg ggaaatgaca aatggtccaa
8761 actagttcga agatctccgg agct
//
