| 质粒名称: | pFA6a-GFP(S65T)-His3MX6 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 41598 |
| 存储实验室: | John Pringle |
|
载体骨架基本信息 |
|
| 载体名称: | pFA6a-kanMX6 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 3938 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 酵母表达；酵母基因组靶向 |
| 筛选标记: | HIS3 |
| 高拷贝/低拷贝: | 未知 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | HIS3 |
| 物种来源: | -- |
| 插入大小: | 2345 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | TEF |
| 标签/融合蛋白: |
GFP（S65T）（N端子插入）
酵母ADH1终止剂（插入物上的N端）
A.棉型翻译延伸因子1a基因启动子（插入端N端）
A.棉型翻译延伸因子1a基因终止子（插入端C端）
|
| 克隆方法: | 限制性酶插入 |
| 5' 克隆位点: | PacI(未破坏) |
| 3' 克隆位点: | PmeI(未破坏) |
| 5' 测序引物 | SP6 |
| 3' 测序引物 | T7 |
|
文献引用方法 |
|
| 材料和方法部分: | pFA6a-GFP(S65T)-His3MX6 was a gift from John Pringle (Addgene plasmid # 41598 ; http://n2t.net/addgene:41598 ; RRID:Addgene_41598) |
| 参考文献部分: |
Additional modules for versatile and economical PCR-based gene deletion and modification in Saccharomyces cerevisiae. Longtine MS, McKenzie A 3rd, Demarini DJ, Shah NG, Wach A, Brachat A, Philippsen P, Pringle JR. Yeast. 1998 Jul;14(10):953-61. 10.1002/(SICI)1097-0061(199807)14:10<953::AID-YEA293>3.0.CO;2-U PubMed 9717241 |
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
-His3MX6质粒图谱.png)
载体简介
载体序列
LOCUS pFA6a-GFP(S65T)-His3MX6 5546 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pFA6a-GFP(S65T)-His3MX6
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5546)
AUTHORS Longtine MS, McKenzie A 3rd, Demarini DJ, Shah NG, Wach A, Brachat
A, Philippsen P, Pringle JR
TITLE Additional modules for versatile and economical PCR-based gene
deletion and modification in Saccharomyces cerevisiae.
JOURNAL Yeast. 1998 Jul;14(10):953-61.
PUBMED 9717241
REFERENCE 2 (bases 1 to 5546)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5546
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 61..774
/codon_start=1
/product="Aequoria victoria green fluorescent protein"
/label=GFP
/translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
ICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
VTAAGITHGMDELYK"
primer_bind complement(88..116)
/label=GFP-R
/note="GFP, reverse primer. Does NOT anneal to EGFP"
primer_bind 711..731
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
terminator 795..982
/gene="S. cerevisiae ADH1"
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
primer_bind complement(850..874)
/label=pHybLex-R
/note="ADH terminator, reverse primer"
promoter 1057..1400
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
terminator 2103..2300
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
primer_bind complement(2404..2423)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(2405..2423)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2510..2527)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2681..3269)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2761..2780)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3440..4300)
/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 4063..4082
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4301..4405)
/gene="bla"
/label=AmpR promoter
mobile_element 4466..5233
/mobile_element_type="insertion sequence:IS1"
/note="prokaryotic transposable element"
primer_bind 5250..5268
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(5306..5328)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 5428..5447
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 5528..5546
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 5528..5545
/label=SP6
/note="SP6 promoter, forward primer"
ORIGIN
1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
61 agtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat
121 gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa
181 cttaccctta aatttatttg cactactgga aaactacctg ttccatggcc aacacttgtc
241 actactttca cttatggtgt tcaatgcttt tcaagatacc cagatcatat gaaacggcat
301 gactttttca agagtgccat gcccgaaggt tatgtacagg aaagaactat atttttcaaa
361 gatgacggga actacaagac acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat
421 agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg acacaaattg
481 gaatacaact ataactcaca caatgtatac atcatggcag acaaacaaaa gaatggaatc
541 aaagttaact tcaaaattag acacaacatt gaagatggaa gcgttcaact agcagaccat
601 tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg
661 tccacacaat ctgccctttc gaaagatccc aacgaaaaga gagaccacat ggtccttctt
721 gagtttgtaa cagctgctgg gattacacat ggcatggatg aactatacaa atagggcgcg
781 ccacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa
841 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat
901 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagtat gaggtcgctc
961 ttattgacca cacctctacc ggcagatccg ctagggataa cagggtaata tagatctgtt
1021 tagcttgcct cgtccccgcc gggtcacccg gccagcgaca tggaggccca gaataccctc
1081 cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag
1141 cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga
1201 agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc cctcacagac
1261 gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca
1321 ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc
1381 acatccgaac ataaacaacc atggcagaac cagcccaaaa aaagcaaaaa caaactgttc
1441 aggagcgcaa ggcgtttatc tcccgtatca ctaatgaaac taaaattcaa atcgctattt
1501 cgctgaatgg tggttatatt caaataaaag attcgattct tcctgcaaag aaggatgacg
1561 atgtagcttc ccaagctact cagtcacagg tcatcgatat tcacacaggt gttggctttt
1621 tggatcatat gatccatgcg ttggcaaaac actctggttg gtctcttatt gttgaatgta
1681 ttggtgacct gcacattgac gatcaccata ctaccgaaga ttgcggtatc gcattagggc
1741 aagcgttcaa agaagcaatg ggtgctgtcc gtggtgtaaa aagattcggt actgggttcg
1801 caccattgga tgaggcgcta tcacgtgccg tagtcgattt atctagtaga ccatttgctg
1861 taatcgacct tggattgaag agagagatga ttggtgattt atccactgaa atgattccac
1921 actttttgga aagtttcgcg gaggcggcca gaattacttt gcatgttgat tgtctgagag
1981 gtttcaacga tcaccacaga agtgagagtg cgttcaaggc tttggctgtt gccataagag
2041 aagctatttc tagcaatggc accaatgacg ttccctcaac caaaggtgtt ttgatgtgaa
2101 gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata
2161 ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca
2221 tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt
2281 gaatgctggt cgctatactg ctgtcgattc gatactaacg ccgccatcca gtttaaacga
2341 gctcgaattc atcgatgata tcagatccac tagtggccta tgcggccgcg gatctgccgg
2401 tctccctata gtgagtcgta ttaatttcga taagccaggt taacctgcat taatgaatcg
2461 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
2521 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
2581 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
2641 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
2701 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
2761 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
2821 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
2881 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
2941 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
3001 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
3061 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
3121 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
3181 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
3241 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
3301 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
3361 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
3421 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
3481 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
3541 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
3601 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
3661 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
3721 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
3781 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
3841 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
3901 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
3961 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
4021 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
4081 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
4141 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
4201 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
4261 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
4321 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
4381 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
4441 aaaccattat tatcatgaca ttaacggtaa tgactccaac ttattgatag tgttttatgt
4501 tcagataatg cccgatgact ttgtcatgca gctccaccga ttttgagaac gacagcgact
4561 tccgtcccag ccgtgccagg tgctgcctca gattcaggtt atgccgctca attcgctgcg
4621 tatatcgctt gctgattacg tgcagctttc ccttcaggcg ggattcatac agcggccagc
4681 catccgtcat ccatatcacc acgtcaaagg gtgacagcag gctcataaga cgccccagcg
4741 tcgccatagt gcgttcaccg aatacgtgcg caacaaccgt cttccggaga ctgtcatacg
4801 cgtaaaacag ccagcgctgg cgcgatttag ccccgacata gccccactgt tcgtccattt
4861 ccgcgcagac gatgacgtca ctgcccggct gtatgcgcga ggttaccgac tgcggcctga
4921 gttttttaag tgacgtaaaa tcgtgttgag gccaacgccc ataatgcggg ctgttgcccg
4981 gcatccaacg ccattcatgg ccatatcaat gattttctgg tgcgtaccgg gttgagaagc
5041 ggtgtaagtg aactgcagtt gccatgtttt acggcagtga gagcagagat agcgctgatg
5101 tccggcggtg cttttgccgt tacgcaccac cccgtcagta gctgaacagg agggacagct
5161 gatagaaaca gaagccactg gagcacctca aaaacaccat catacactaa atcagtaagt
5221 tggcagcatc accgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
5281 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
5341 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
5401 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
5461 atatggacat attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac
5521 acatacgatt taggtgacac tataga
//
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