| 质粒名称: | pCU-MET3-GFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 45337 |
| 存储实验室: | Brendan Cormack |
|
载体骨架基本信息 |
|
| 载体名称: | pGRB2.3 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 7009 bp |
| 载体修饰: | 用白色念珠菌中MET3启动子替换pGRB2.3中的PGK1启动子（使用SacI/XbaI限制性消化） |
| 载体类型: | 白色念珠菌使用载体。 |
| 筛选标记: | URA3 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
| 相关菌种: | 白色念珠菌SC5314 |
|
插入基因信息 |
|
| 插入基因名称: | yeGFP,GFP |
| 物种来源: | Synthetic |
| 插入大小: | 717 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | MET3 |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI（销毁后未知） |
| 3' 克隆位点: | SalI （销毁后未知） |
| 5' 测序引物 | M13R |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pCU-MET3-GFP was a gift from Brendan Cormack (Addgene plasmid # 45337 ; http://n2t.net/addgene:45337 ; RRID:Addgene_45337) |
| 参考文献部分: | Expression plasmids for use in Candida glabrata. Zordan RE, Ren Y, Pan SJ, Rotondo G, De Las Penas A, Iluore J, Cormack BP. G3 (Bethesda). 2013 Oct 3;3(10):1675-86. doi: 10.1534/g3.113.006908. 10.1534/g3.113.006908 PubMed 23934995 |
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 pCU-MET3-GFP 7009 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCU-MET3-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7009)
AUTHORS Zordan RE, Ren Y, Pan SJ, Rotondo G, De Las Penas A, Iluore J,
Cormack BP
TITLE Expression plasmids for use in Candida glabrata.
JOURNAL G3 (Bethesda). 2013 Oct 3;3(10):1675-86. doi: 10.1534/g3.113.006908.
PUBMED 23934995
REFERENCE 2 (bases 1 to 7009)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7009
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(10..28)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 666..737
/gene="bla"
/label=AmpR promoter
CDS 738..1598
/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(956..975)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1769..2357
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2258..2277
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2511..2528
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 2645..2666
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2681..2711
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2719..2735
/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 2724..2746
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 2743..2759
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2743..2759
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 2778..2798
/label=T3
/note="T3 promoter, forward primer"
promoter 2780..2798
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind 3843..3859
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3843..3859
/label=pBluescriptSK
/note="For pBluescript vector"
CDS 3879..4595
/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"
primer_bind complement(5025..5044)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(5026..5044)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(5054..5071)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(5054..5070)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5063..5085)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 5211..5666
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5298..5317)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5508..5529
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(5797..6600)
/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(6601..6816)
/gene="S. cerevisiae URA3"
/label=URA3 promoter
primer_bind complement(6840..6859)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6959..6981
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
ORIGIN
1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
61 cttagcaggt tagactaaca tgcaaaaagt ttatattcat ttttgtagtt attcttgttt
121 tcattaggat gattacacac tgtggatgtg gattaatcga tcattgttaa tttagtgtag
181 ctactaccat ttcaaacaaa acgtaaattc ttgcagtcgt actggatctg tgaatctatt
241 agtatatatg aattaaagta gcttgtacat attattctgt tgaatcatat cgcagagcat
301 tggttgaaat ccaaaatata aaaatgtaat atcacaaaaa ataatactaa ttctaacatt
361 aatgggtcag atttttagtg aatacttaaa tttataatat cgtctattta agctagcaaa
421 tggaacaaca tttaaagtaa gaacatcata tctacatgaa aatgtatatt tcaatctgac
481 taataacgca gagcacatct ttcagtgatg tctgtcacat gatcaaaaag aattgtattt
541 aaataattca taataaaagc ttaaaaaatt acaataatga aaataaagta aataatgaca
601 tgggtaagag ttccgaaata gaatcttagt gtacaaaaca aaaattgcat cattagagat
661 ccccacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
721 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
781 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
841 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
901 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
961 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
1021 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
1081 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
1141 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
1201 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
1261 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
1321 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
1381 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
1441 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
1501 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
1561 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
1621 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
1681 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
1741 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
1801 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
1861 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
1921 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
1981 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
2041 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
2101 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
2161 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
2221 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
2281 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
2341 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
2401 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
2461 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
2521 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
2581 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
2641 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
2701 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
2761 atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg gagctcatac
2821 cagttacaat tagtattaca atggtttacc aagtacaaat tatgtaatat ggttataatt
2881 ggcttatatg tatggaggta gggaactgca ggttattgtt ctgttaaact accagtaatg
2941 caagacggct aagtcatatg actgccactt tgtcaatcct gaagaaaatg acaatagaaa
3001 gtgatataat acacgtgata tgcgtaggat gggacccaaa gatagccaca tcacatgatc
3061 ttcaaaaacc ccgtcaaaaa ggaataaatc gaaggaaaaa aaatgccacg tgactttgat
3121 agtcaaaatc aggttatatt actatacgat cccccacaca taacctttac cacacaccgc
3181 acgggctgga cttatcctat tagaaagcgg tgcagccagg gccaagaaaa cgcgaacgac
3241 gcagaaaaaa cgtcaagcaa aaaaactgtg gtgttattta aagcatacaa tttcctgcct
3301 ccctttaatg ctacggggga tctagcaaat gggaaaatat cacatgactt tggctagaag
3361 gggtgggaaa acagatattt tagtcacatt gtttgatttc acgtactaca cgatacacta
3421 cactacacaa tacactgcaa tacactacac tgtatggttc cctccggccc tggatatgct
3481 agcgaaggat cccaagccca tcgaggaaat cattcaagag catctgcagg tctcaaatac
3541 actaagtcca aacacaaaca gcactgagct aatccaagac accaattgca tgccttctca
3601 attagaacag tttattacac agcttataac tgtgcatctt gccattcttt tcgagatagc
3661 catttgcatt tgattaacat gcttagctcg ttctcaagcc acagtaaaat ggattgcctt
3721 ttgagtttcc atcgttgtat aaataggcac tcattcctat agccttgctc tttggtcttg
3781 tctctgtaat ggtaatagct gtaagtcagg atataacact ccagaaaaga aacacctaac
3841 aatctagaac tagtggatcc cccgggctgc aggaattcat gtctaaaggt gaagaattat
3901 tcactggtgt tgtcccaatt ttggttgaat tagatggtga tgttaatggt cacaaatttt
3961 ctgtctccgg tgaaggtgaa ggtgatgcta cttacggtaa attgacctta aaatttattt
4021 gtactactgg taaattgccg gttccatggc caaccttagt cactactttc ggttatggtg
4081 ttcaatgttt tgctagatac ccagatcata tgaaacaaca tgactttttc aagtctgcca
4141 tgccagaagg ttatgttcaa gaaagaacta tttttttcaa agatgacggt aactacaaga
4201 ccagagctga agtcaagttt gaaggtgata ccttagttaa tagaatcgaa ttaaaaggta
4261 ttgattttaa agaagatggt aacattttag gtcacaaatt ggaatacaac tataactctc
4321 acaatgttta catcatggct gacaaacaaa agaatggtat caaagttaac ttcaaaatta
4381 gacacaacat tgaagatggt tctgttcaat tagctgacca ttatcaacaa aatactccaa
4441 ttggtgatgg tccagtcttg ttaccagaca accattactt atccactcaa tctgccttat
4501 ccaaagatcc aaacgaaaag agagaccaca tggtcttgtt agaatttgtt actgctgctg
4561 gtattaccca tggtatggat gaattgtaca aataactgca ggtcgacctc gagaacacaa
4621 cccacagcta ccaccatcaa caatatttat atatataacg tacacataga aatcacacaa
4681 acagagtatt tattcttaac tacatgaact accatcagac cgtctgggcc actatataat
4741 gtgccattca taaacgtgat cactttacgt agcaggcaac cccaggtgaa aatttttcag
4801 cgagctgcca gattgtcagg tgaaaaactg aaaaaaactt ctgggcgatg agcttgtggc
4861 gggaaattaa gtatataaag cagttagttt cttctgcttc ttgtggttct gggtattctt
4921 gtaatatttg gaagaataga aaatcaagac tactgccttt cttttcatat tatagaggaa
4981 gaaatacgca cgaacacgat atagaggtaa aggtacccaa ttcgccctat agtgagtcgt
5041 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
5101 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
5161 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg
5221 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
5281 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
5341 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
5401 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
5461 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
5521 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
5581 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
5641 taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc
5701 ggtatttcac accgcatagg gtaataactg atataattaa attgaagctc taatttgtga
5761 gtttagtata catgcattta cttataatac agttttttag ttttgctggc cgcatcttct
5821 caaatatgct tcccagcctg cttttctgta acgttcaccc tctaccttag catcccttcc
5881 ctttgcaaat agtcctcttc caacaataat aatgtcagat cctgtagaga ccacatcatc
5941 cacggttcta tactgttgac ccaatgcgtc tcccttgtca tctaaaccca caccgggtgt
6001 cataatcaac caatcgtaac cttcatctct tccacccatg tctctttgag caataaagcc
6061 gataacaaaa tctttgtcgc tcttcgcaat gtcaacagta cccttagtat attctccagt
6121 agatagggag cccttgcatg acaattctgc taacatcaaa aggcctctag gttcctttgt
6181 tacttcttct gccgcctgct tcaaaccgct aacaatacct gggcccacca caccgtgtgc
6241 attcgtaatg tctgcccatt ctgctattct gtatacaccc gcagagtact gcaatttgac
6301 tgtattacca atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt acttggcgga
6361 taatgccttt agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga tatccacatg
6421 tgtttttagt aaacaaattt tgggacctaa tgcttcaact aactccagta attccttggt
6481 ggtacgaaca tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga tattaaatag
6541 cttggcagca acaggactag gatgagtagc agcacgttcc ttatatgtag ctttcgacat
6601 gatttatctt cgtttcctgc aggtttttgt tctgtgcagt tgggttaaga atactgggca
6661 atttcatgtt tcttcaacac tacatatgcg tatatatacc aatctaagtc tgtgctcctt
6721 ccttcgttct tccttctgtt cggagattac cgaatcaaaa aaatttcaag gaaaccgaaa
6781 tcaaaaaaaa gaataaaaaa aaaatgatga attgaaaagg tggtatggtg cactctcagt
6841 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
6901 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
6961 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcga
//
载体图谱质粒图谱pdf版和相关资料下载
载体质粒应用举例
- 上一篇：T7opt in pCAGGS
- 下一篇：pFZ19
