| 出品人: | Irene Castano Navarro（2015） |
|---|---|
| 载体名称: | pYC25 |
| 质粒类型: | 酿酒酵母载体；复制型载体；过表达载体 |
| 高拷贝/低拷贝: | 高拷贝 |
| 克隆方法: | 限制性内切酶，多克隆位点 |
| 启动子: | TEF1 |
| 载体大小: | 6418 bp |
| 5' 测序引物及序列: | M13/pUC fwd primer 5’ CCAGTCACGACGTTGTAAAACG 3’ |
| 3' 测序引物及序列: | GFP-R primer 5’ ACCATCTAATTCAACAAGAATTGGGACAAC 3’ |
| 载体标签: | 黄色荧光蛋白YFP tag |
| 载体抗性: | 氨苄青霉素 |
| 筛选标记: | URA3 |
| 克隆菌株: | DH10B感受态细胞 ,30°C |
| 宿主细胞（系）: | 出芽酵母（酿酒酵母）、白色念珠菌 |
| 备注: | -- |
| 产品目录号: | #63914 |
| 稳定性: | 瞬表达 |
| 组成型/诱导型: | 组成型 |
| 病毒/非病毒: | 非病毒 |
买家导航
pYC25载体质粒基本信息
pYC25质粒图谱载体图谱和pYC25载体序列质粒序列多克隆位点信息
pYC25质粒载体简介
pYC25质粒序列载体序列
LOCUS Exported 6418 bp ds-DNA circular SYN 14-AUG-2016
DEFINITION synthetic circular DNA
KEYWORDS pYC25
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6418)
AUTHORS Biofeng Lab.
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..6418
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 666..737
/gene="bla"
/note="AmpR promoter"
CDS 738..1598
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 1753..2372
/direction=RIGHT
/note="pBR322"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 2681..2711
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 2719..2735
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/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 2743..2759
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 2780..2798
/note="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 2816..3214
/gene="S. cerevisiae TEF1"
/note="TEF1 promoter"
/note="promoter for EF-1-alpha"
misc_feature 3218..3235
/note="MCS"
primer_bind 3218..3234
/note="SK primer"
/note="common sequencing primer, one of multiple similar
variants"
CDS 3238..3954
/codon_start=1
/note="YFP"
/translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
LICTTGKLPVPWPTLVTTLGYGLMCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
primer_bind complement(3993..4009)
/note="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(4424..4442)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4452..4468)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 4613..5068
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(5199..6002)
/codon_start=1
/gene="S. cerevisiae URA3"
/product="orotidine-5'-phosphate decarboxylase, required
for uracil biosynthesis"
/note="URA3"
/note="yeast auxotrophic marker, counterselectable with
5-fluoroorotic acid (5-FOA)"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
promoter complement(6003..6218)
/gene="S. cerevisiae URA3"
/note="URA3 promoter"
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 actgtccttc
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 gagttacctc actcattagg caccccaggc tttacacttt atgcttccgg
2701 ctcctatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
2761 atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg gagctcatag
2821 cttcaaaatg tttctactcc ttttttactc ttccagattt tctcggactc cgcgcatcgc
2881 cgtaccactt caaaacaccc aagcacagca tactaaattt cccctctttc ttcctctagg
2941 gtgtcgttaa ttacccgtac taaaggtttg gaaaagaaaa aagagaccgc ctcgtttctt
3001 tttcttcgtc gaaaaaggca ataaaaattt ttatcacgtt tctttttctt gaaaattttt
3061 tttttgattt ttttctcttt cgatgacctc ccattgatat ttaagttaat aaacggtctt
3121 caatttctca agtttcagtt tcatttttct tgttctatta caactttttt tacttcttgc
3181 tcattagaaa gaaagcatag caatctaatc taagttttct agaactagtg gatcctaatg
3241 agtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat
3301 gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa
3361 cttaccctta aattgatttg cactactgga aaactacctg ttccatggcc aacacttgtc
3421 actactttgg gttatggtct aatgtgcttt gctagatacc cagatcatat gaaacggcat
3481 gactttttca agagtgccat gcccgaaggt tatgtacagg aaagaactat atttttcaaa
3541 gatgacggga actacaagac acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat
3601 agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg acacaaattg
3661 gaatacaact ataactcaca caatgtatac atcactgcag acaaacaaaa gaatggaatc
3721 aaagctaact tcaaaattag acacaacatt gaagatggag gtgttcaact agcagaccat
3781 tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg
3841 tcctatcaat ctgccctttc gaaagatccc aacgaaaaga gagaccacat ggtccttctt
3901 gagtttgtaa cagctgctgg gattacacat ggcatggatg aactatacaa atagagatcc
3961 cccgggctgc aggaattcga tatcaagctt atcgataccg tcgacctcga gaacacaacc
4021 cacagctacc accatcaaca atatttatat atataacgta cacatagaaa tcacacaaac
4081 agagtattta ttcttaacta catgaactac catcagaccg tctgggccac tatataatgt
4141 gccattcata aacgtgatca ctttacgtag caggcaaccc caggtgaaaa tttttcagcg
4201 agctgccaga ttgtcaggtg aaaaactgaa aaaaacttct gggcgatgag cttgtggcgg
4261 gaaattaagt atataaagca gttagtttct tctgcttctt gtggttctgg gtattcttgt
4321 aatatttgga agaatagaaa atcaagacta ctgcctttct tttcatatta tagaggaaga
4381 aatacgcacg aacacgatat agaggtaaag gtacccaatt cgccctatag tgagtcgtat
4441 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
4501 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
4561 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cgacgcgccc
4621 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
4681 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
4741 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
4801 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
4861 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
4921 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
4981 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
5041 tttaacaaaa tattaacgtt tacaatttcc tgatgcggta ttttctcctt acgcatctgt
5101 gcggtatttc acaccgcata gggtaataac tgatataatt aaattgaagc tctaatttgt
5161 gagtttagta tacatgcatt tacttataat acagtttttt agttttgctg gccgcatctt
5221 ctcaaatatg cttcccagcc tgcttttctg taacgttcac cctctacctt agcatccctt
5281 ccctttgcaa atagtcctct tccaacaata ataatgtcag atcctgtaga gaccacatca
5341 tccacggttc tatactgttg acccaatgcg tctcccttgt catctaaacc cacaccgggt
5401 gtcataatca accaatcgta accttcatct cttccaccca tgtctctttg agcaataaag
5461 ccgataacaa aatctttgtc gctcttcgca atgtcaacag tacccttagt atattctcca
5521 gtagataggg agcccttgca tgacaattct gctaacatca aaaggcctct aggttccttt
5581 gttacttctt ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt
5641 gcattcgtaa tgtctgccca ttctgctatt ctgtatacac ccgcagagta ctgcaatttg
5701 actgtattac caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg
5761 gataatgcct ttagcggctt aactgtgccc tccatggaaa aatcagtcaa gatatccaca
5821 tgtgttttta gtaaacaaat tttgggacct aatgcttcaa ctaactccag taattccttg
5881 gtggtacgaa catccaatga agcacacaag tttgtttgct tttcgtgcat gatattaaat
5941 agcttggcag caacaggact aggatgagta gcagcacgtt ccttatatgt agctttcgac
6001 atgatttatc ttcgtttcct gcaggttttt gttctgtgca gttgggttaa gaatactggg
6061 caatttcatg tttcttcaac actacatatg cgtatatata ccaatctaag tctgtgctcc
6121 ttccttcgtt cttccttctg ttcggagatt accgaatcaa aaaaatttca aagaaaccga
6181 aatcaaaaaa aagaataaaa aaaaaatgat gaattgaatt gaaaagctgt ggtatggtgc
6241 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
6301 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
6361 accgtctccg ggaggtgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcga
//
