| 质粒名称: | PTEF1-yEGFPCLN2PEST-pRS406 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 64406 |
| 存储实验室: | Claudia Vickers |
|
载体骨架基本信息 |
|
| 载体名称: | GFPCLN2PEST-pRS406 |
| 载体出品公司: | Mateus and Avery 2000 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | destabilized GFP |
| 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: | TEF1 promoter |
| 物种来源: | S. cerevisiae (budding yeast) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | TEF1 promoter |
| 标签/融合蛋白: | yEGFPCLN2PEST（失稳酵母EGFP）（主链上的C端） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | XhoI (未破坏) |
| 3’克隆位点: | EcoRI(未破坏) |
| 5’测序引物 | M13pUC-fwd |
| 3’测序引物 | yGFP-R |
|
文献引用方法 |
|
| 材料和方法部分: | PTEF1-yEGFPCLN2PEST-pRS406 was a gift from Claudia Vickers (Addgene plasmid # 64406 ; http://n2t.net/addgene:64406 ; RRID:Addgene_64406) |
| 参考文献部分: | Dynamic regulation of gene expression using sucrose responsive promoters and RNA interference in Saccharomyces cerevisiae. Williams TC, Espinosa MI, Nielsen LK, Vickers CE. Microb Cell Fact. 2015 Apr 1;14:43. doi: 10.1186/s12934-015-0223-7. 10.1186/s12934-015-0223-7 [pii] PubMed 25886317 |
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 PTEF1-yEGFPCLN2PEST-pRS406 6235 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION TEF promoter driven GFP expression.
ACCESSION .
VERSION .
KEYWORDS PTEF1-yEGFPCLN2PEST-pRS406
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6235)
AUTHORS Williams TC, Espinosa MI, Nielsen LK, Vickers CE
TITLE Dynamic regulation of gene expression using sucrose responsive
promoters and RNA interference in Saccharomyces cerevisiae.
JOURNAL Microb Cell Fact. 2015 Apr 1;14:43. doi: 10.1186/s12934-015-0223-7.
PUBMED 25886317
REFERENCE 2 (bases 1 to 6235)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6235
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(29..51)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 151..170
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 201..416
/gene="S. cerevisiae URA3"
/label=URA3 promoter
CDS 417..1220
/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"
rep_origin complement(1351..1806)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1488..1509)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 1700..1719
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1936..1958
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 1950..1967
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 1951..1967
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 1977..1996
/label=T7
/note="T7 promoter, forward primer"
promoter 1977..1995
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 2026..2432
/gene="S. cerevisiae TEF1"
/label=TEF1 promoter
/note="promoter for EF-1-alpha"
CDS 2456..3169
/codon_start=1
/product="yeast-enhanced green fluorescent protein"
/label=yeGFP
/note="codons optimized for translation in S. cerevisiae
and C. albicans"
/translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
terminator 3727..3914
/gene="S. cerevisiae ADH1"
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
primer_bind complement(3782..3806)
/label=pHybLex-R
/note="ADH terminator, reverse primer"
primer_bind complement(3922..3938)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3922..3938)
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(3975..3995)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(3975..3993)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(4014..4030)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4014..4030)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4027..4049)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4038..4054
/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(4062..4092)
/label=lac promoter
/note="promoter for the E. coli lac operon"
primer_bind complement(4245..4262)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4416..5004)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4496..4515)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5175..6035)
/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 5798..5817
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6036..6140)
/gene="bla"
/label=AmpR promoter
primer_bind 6208..6226
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
181 accataccac agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc
241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
301 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
361 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt
421 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
481 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
541 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
601 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
661 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca
721 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
781 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
841 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
901 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
961 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
1021 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg
1081 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
1141 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
1201 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
1261 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac
1321 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat
1381 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
1441 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
1501 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
1561 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
1621 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
1681 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
1741 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
1801 gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
1861 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
1921 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat
1981 acgactcact atagggcgaa ttgggtaccg ggccccccct cgaggcacac accatagctt
2041 caaaatgttt ctactccttt tttactcttc cagattttct cggactccgc gcatcgccgt
2101 accacttcaa aacacccaag cacagcatac taaatttccc ctctttcttc ctctagggtg
2161 tcgttaatta cccgtactaa aggtttggaa aagaaaaaag agaccgcctc gtttcttttt
2221 cttcgtcgaa aaaggcaata aaaattttta tcacgtttct ttttcttgaa aatttttttt
2281 tttgattttt ttctctttcg atgacctccc attgatattt aagttaataa acggtcttca
2341 atttctcaag tttcagtttc atttttcttg ttctattaca acttttttta cttcttgctc
2401 attagaaaga aagcatagca atctaatcta agttttaatt acaagaattc attaaatgtc
2461 taaaggtgaa gaattattca ctggtgttgt cccaattttg gttgaattag atggtgatgt
2521 taatggtcac aaattttctg tctccggtga aggtgaaggt gatgctactt acggtaaatt
2581 gaccttaaaa tttatttgta ctactggtaa attgccagtt ccatggccaa ccttagtcac
2641 tactttcggt tatggtgttc aatgttttgc gagataccca gatcatatga aacaacatga
2701 ctttttcaag tctgccatgc cagaaggtta tgttcaagaa agaactattt ttttcaaaga
2761 tgacggtaac tacaagacca gagctgaagt caagtttgaa ggtgatacct tagttaatag
2821 aatcgaatta aaaggtattg attttaaaga agatggtaac attttaggtc acaaattgga
2881 atacaactat aactctcaca atgtttacat catggctgac aaacaaaaga atggtatcaa
2941 agttaacttc aaaattagac acaacattga agatggttct gttcaattag ctgaccatta
3001 tcaacaaaat actccaattg gtgatggtcc agtcttgtta ccagacaacc attacttatc
3061 cactcaatct gccttatcca aagatccaaa cgaaaagaga gaccacatgg tcttgttaga
3121 atttgttact gctgctggta ttacccatgg tatggatgaa ttgtacaaag catccaactt
3181 gaacatttcg agaaagctta ccatatcaac cccatcatgc tctttcgaaa attcaaatag
3241 cacatccatt ccttcgcccg cttcctcatc tcaaagccac actccaatga gaaacatgag
3301 ctcactctct gataacagcg ttttcagccg gaatatggaa caatcatcac caatcactcc
3361 aagtatgtac caatttggtc agcagcagtc aaacagtata tgtggtagca ccgttagtgt
3421 gaatagtctg gtgaatacaa ataacaaaca aaggatctac gaacaaatca cgggtcctaa
3481 cagcaataac gcaaccaatg attatattga tttgctaaac ctaaatgagt ctaacaagga
3541 aaaccaaaat cccgcaacgg cgcattacct caatgggggc ccacccaaga caagcttcat
3601 taaccatgga atgttcccct cgccaactgg gaccataaat agcggtaaat ctagcagtgc
3661 ctcatcttta atttcttttg gtatgggcaa tacccaagta atatagggcg cgccacttct
3721 aaataagcga atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata
3781 agtgtataca aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt
3841 aactctttcc tgtaggtcag gttgctttct caggtatagt atgaggtcgc tcttattgac
3901 cacacctcta ccggcagatc ggatccacta gttctagagc ggccgccacc gcggtggagc
3961 tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
4021 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatagg agccggaagc
4081 ataaagtgta aagcctgggg tgcctaatga gtgaggtaac tcacattaat tgcgttgcgc
4141 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
4201 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
4261 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
4321 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
4381 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
4441 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
4501 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
4561 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
4621 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
4681 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
4741 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
4801 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
4861 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
4921 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
4981 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
5041 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
5101 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
5161 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
5221 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
5281 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
5341 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
5401 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
5461 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
5521 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
5581 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
5641 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
5701 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
5761 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
5821 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
5881 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
5941 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
6001 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
6061 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
6121 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
6181 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtc
//
