| 质粒名称: | pDRf1-GW |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 36026 |
| 存储实验室: | Wolf Frommer, Dominique Loque |
|
载体骨架基本信息 |
|
| 载体名称: | pDRf1 |
| 载体出品公司: | -- |
| 空载体大小: | 9100 bp |
| 完整质粒大小: | -- |
| 载体修饰: | 在msc中插入invitrogen的GW克隆盒 |
| 载体类型: | 酵母表达 |
| 筛选标记: | URA3 |
| 载体抗性: | 氨苄霉素100 μg/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: | -- |
| 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: | pDRf1-GW was a gift from Wolf Frommer & Dominique Loque (Addgene plasmid # 36026 ; http://n2t.net/addgene:36026 ; RRID:Addgene_36026) |
| 参考文献部分: | A cytosolic trans-activation domain essential for ammonium uptake. Loque D, Lalonde S, Looger LL, von Wiren N, Frommer WB. Nature. 2007 Mar 8;446(7132):195-8. Epub 2007 Feb 11. 10.1038/nature05579 PubMed 17293878 |
买家导航
pDRf1GW载体基本信息
pDRf1GW载体图谱质粒图谱和多克隆位点信息
pDRf1GW载体简介
pDRf1GW载体序列
LOCUS pDRf1-GW 8606 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pDRf1-GW
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8606)
AUTHORS Loque D, Lalonde S, Looger LL, von Wiren N, Frommer WB
TITLE A cytosolic trans-activation domain essential for ammonium uptake.
JOURNAL Nature. 2007 Mar 8;446(7132):195-8. Epub 2007 Feb 11.
PUBMED 17293878
REFERENCE 2 (bases 1 to 8606)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8606
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 22..146
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 183..213
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
primer_bind complement(333..352)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
CDS 1271..1576
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
V?RELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
primer_bind 1455..1474
/label=ccdB-fwd
/note="ccdB gene, forward primer"
protein_bind complement(1617..1741)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(1803..1822)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1804..1822)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
terminator 1970..2157
/gene="S. cerevisiae ADH1"
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
primer_bind complement(2025..2049)
/label=pHybLex-R
/note="ADH terminator, reverse primer"
primer_bind complement(2169..2186)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(2169..2185)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2178..2200)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 2621..3967
/label=2u ori
/note="yeast 2u plasmid origin of replication"
CDS complement(4078..4881)
/codon_start=1
/gene="S. cerevisiae URA3"
/product="orotidine-5'-phosphate decarboxylase, required
for uracil biosynthesis"
/label=URA3
/note="yeast auxotrophic marker, counterselectable with
5-fluoroorotic acid (5-FOA)"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
promoter complement(4882..5097)
/gene="S. cerevisiae URA3"
/label=URA3 promoter
primer_bind complement(5122..5140)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5208..5312
/gene="bla"
/label=AmpR promoter
CDS 5313..6173
/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 complement(5531..5550)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6344..6932
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6833..6852
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7086..7103
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 7220..7241
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 7256..7286
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7294..7310
/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)."
primer_bind 7299..7321
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 7318..7334
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 7318..7334
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
rep_origin complement(7364..7819)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7501..7522)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 7713..7732
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 actagtgaat tcgcggccat cacaagtttg tacaaaaaag ctgaacgaga aacgtaaaat
61 gatataaata tcaatatatt aaattagatt ttgcataaaa aacagactac ataatactgt
121 aaaacacaac atatccagtc actatggcgg cccctcgagg ggccgcatta ggcaccccag
181 gctttacact ttatgcttcc ggctcgtata atgtgtggat tttgagttag gatccgtcga
241 gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg
301 atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta
361 cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata
421 agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg
481 aattccgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt
541 acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg
601 atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg
661 cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga
721 gtttcaccag tttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc
781 accatgggca aatattatac gcaagggcga caaggtgctg atgccgctgg cgattcaggt
841 tcatcatgcc gtttgtgatg gcttccatgt cgggcagaat gcttaatgaa ttacaacagt
901 actgcgatga gtggcagggc ggggcgtaan cgcgtggatc cggcttacta aaagccagat
961 aacagtatgc gtatttgcgc gctgattttt gcggtataag aatatatact gatatgtata
1021 cccgaagtat gtcaaaaaga ggtatgctat gaagcagcgt attacagnga cagttgacag
1081 cgacagctat cagttgctca aggcatatat gatgtcaata tctccggtct ggtaagcaca
1141 accatgcaga atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa
1201 gggatggctg aggtcgcccg gtttattgaa atgaacggct cttttgctga cgagaacagg
1261 ggctggtgaa atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt
1321 tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc
1381 cagtgcacgt ctgctgtcag ataaagtcnc ccgtgaactt tacccggtgg tgcatatcgg
1441 ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg
1501 ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat
1561 gttctgggga atataaatgt caggctccct tatacacagc cagtctgcag gtcgaccata
1621 gtgactggat atgttgtgtt ttacagtatt atgtagtctg ttttttatgc aaaatctaat
1681 ttaatatatt gatatttata tcattttacg tttctcgttc agctttcttg tacaaagtgg
1741 tgtagttaat tcatgatggc cgctgcaggt cgacctcgag ggggggcccg gtacccaatt
1801 cgccctatag tgagtcgtat tacgcgcgga tccagctttg gacttcttcg ccagaggttt
1861 ggtcaagtct ccaatcaagg ttgtcggctt gtctaccttg ccagaaattt acgaaaagat
1921 ggaaaagggt caaatcgttg gtagatacgt tgttgacact tctaaataag cgaatttctt
1981 atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta
2041 aagtgactct taggttttaa aacganaatt cttattcttg agtaactctt tcctgtaggt
2101 caggttgctt tctcaggtat agcatgaggt cgctcttatt gaccacacct ctaccggcat
2161 gccaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
2221 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
2281 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
2341 tttctcctta cgcatctgtg cggtatttca caccgcataa tcggatcgta cttgttaccc
2401 atcattgaat tttgaacatc cgaacctggg agttttccct gaaacagata gtatatttga
2461 acctgtataa taatatatag tctagcgctt tacggaagac aatgtatgta tttcggttcc
2521 tggagaaact attgcatcta ttgcataggt aatcttgcac gtcgcatccc cggttcattt
2581 tctgcgtttc catcttgcac ttcaatagca tatctttgtt aacgaagcat ctgtgcttca
2641 ttttgtagaa caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg
2701 catttttaca gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc
2761 ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg
2821 agctgcattt ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc
2881 tatacttctt ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca
2941 tcttagatta ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt
3001 gcactgtagg tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata
3061 aaaaaagcct gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt
3121 tttcaagata aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt
3181 gaacagaaag tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt
3241 ctattttgtc tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt
3301 cactctatga atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac
3361 ataaaaaatg tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag
3421 gttatatagg gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt
3481 ggaagcggta ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt
3541 gaaagtgcgt cttcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc
3601 tagctagaga ataggaactt cggaatagga acttcaaagc gtttccgaaa acgagcgctt
3661 ccgaaaatgc aacgcgagct gcgcacatac agctcactgt tcacgtcgca cctatatctg
3721 cgtgttgcct gtatatatat atacatgaga agaacggcat agtgcgtgtt tatgcttaaa
3781 tgcgtactta tatgcgtcta tttatgtagg atgaaaggta gtctagtacc tcctgtgata
3841 ttatcccatt ccatgcgggg tatcgtatgc ttccttcagc actacccttt agctgttcta
3901 tatgctgcca ctcctcaatt ggattagtct catccttcaa tgctatcatt tcctttgata
3961 ttggatcgat ccgatgataa gctgtcaaac atgagaattg ggtaataact gatataatta
4021 aattgaagct ctaatttgtg agtttagtat acatgcattt acttataata cagtttttta
4081 gttttgctgg ccgcatcttc tcaaatatgc ttcccagcct gcttttctgt aacgttcacc
4141 ctctacctta gcatcccttc cctttgcaaa tagtcctctt ccaacaataa taatgtcaga
4201 tcctgtagag accacatcat ccacggttct atactgttga cccaatgcgt ctcccttgtc
4261 atctaaaccc acaccgggtg tcataatcaa ccaatcgtaa ccttcatctc ttccacccat
4321 gtctctttga gcaataaagc cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt
4381 acccttagta tattctccag tagataggga gcccttgcat gacaattctg ctaacatcaa
4441 aaggcctcta ggttcctttg ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc
4501 tgggcccacc acaccgtgtg cattcgtaat gtctgcccat tctgctattc tgtatacacc
4561 cgcagagtac tgcaatttga ctgtattacc aatgtcagca aattttctgt cttcgaagag
4621 taaaaaattg tacttggcgg ataatgcctt tagcggctta actgtgccct ccatggaaaa
4681 atcagtcaag atatccacat gtgtttttag taaacaaatt ttgggaccta atgcttcaac
4741 taactccagt aattccttgg tggtacgaac atccaatgaa gcacacaagt ttgtttgctt
4801 ttcgtgcatg atattaaata gcttggcagc aacaggacta ggatgagtag cagcacgttc
4861 cttatatgta gctttcgaca tgatttatct tcgtttcctg catgtttttg ttctgtgcag
4921 ttgggttaag aatactgggc aatttcatgt ttcttcaaca ctacatatgc gtatatatac
4981 caatctaagt ctgtgctcct tccttcgttc ttccttctgt tcggagatta ccgaatcaaa
5041 aaaatttcaa ggaaaccgaa atcaaaaaaa agaataaaaa aaaaatgatg aattgaaaag
5101 ctaattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
5161 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
5221 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
5281 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
5341 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
5401 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
5461 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
5521 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
5581 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
5641 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
5701 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
5761 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
5821 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
5881 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
5941 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
6001 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
6061 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
6121 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
6181 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
6241 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
6301 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
6361 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
6421 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
6481 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
6541 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
6601 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
6661 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
6721 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
6781 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
6841 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
6901 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
6961 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
7021 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
7081 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
7141 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
7201 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
7261 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
7321 gaaacagcta tgaccatgat tacgccaagc ttaccgcatc aggaaattgt aagcgttaat
7381 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
7441 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
7501 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
7561 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
7621 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
7681 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
7741 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
7801 gcgccgctac agggcgcgtc cattcgccaa gcttcctgaa acggagaaac ataaacaggc
7861 attgctggga tcacccatac atcactctgt tttgcctgac cttttccggt aatttgaaaa
7921 caaacccggt ctcgaagcgg agatccggcg ataattaccg cagaaataaa cccatacacg
7981 agacgtagaa ccagccgcac atggccggag aaactcctgc gagaatttcg taaactcgcg
8041 cgcattgcat ctgtatttcc taatgcggca cttccaggcc tcgatcgaga ccgtttatcc
8101 attgcttttt tgttgtcttt ttccctcgtt cacagaaagt ctgaagaagc tatagtagaa
8161 ctatgagctt tttttgtttc tgttttcctt tttttttttt ttacctctgt ggaaattgtt
8221 actctcacac tctttagttc gtttgtttgt tttgtttatt ccaattatga ccggtgacga
8281 aacgtggtcg atggtgggta ccgcttatgc tcccctccat tagtttcgat tatataaaaa
8341 ggccaaatat tgtattattt tcaaatgtcc tatcattatc gtctaacatc taatttctct
8401 taaatttttt ctctttcttt cctataacac caatagtgaa aatctttttt tcttctatat
8461 ctacaaaaac tttttttttc tatcaacctc gttgataaat tttttcttta acaatcgtta
8521 ataattaatt aattggaaaa taaccatttt ttctctcttt tatacacaca ttcaaaagaa
8581 agaaaaaaaa tataccccag cctcga
//
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