| 质粒名称: | pFA6a-kanMX6-PGAL1-GFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 41614 |
| 存储实验室: | John Pringle |
|
载体骨架基本信息 |
|
| 载体名称: | pFA6a |
| 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: | -- |
| 载体修饰: | -- |
| 载体类型: | 酵母基因组靶向 |
| 筛选标记: | 庆大霉素 |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | GAL1 promoter,GAL1 |
| 物种来源: | S. cerevisiae (budding yeast) |
| 插入大小: | 2807 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: |
A、 棉型翻译延伸因子1a基因终止子（插入子上的N末端）
KanR反向定向（插入件上的N端子）
A、 棉型翻译延伸因子1a基因启动子（插入子N端）
GFP（S65T）（插接件上的C端子）
酵母ADH1终止子（插入件上的C端子）
|
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BglII(未破坏) |
| 3' 克隆位点: | PacI(未破坏) |
| 5' 测序引物 | T7_promoter |
| 3' 测序引物 | Sp6_primer |
|
文献引用方法 |
|
| 材料和方法部分: | pFA6a-kanMX6-PGAL1-GFP was a gift from John Pringle (Addgene plasmid # 41614 ; http://n2t.net/addgene:41614 ; RRID:Addgene_41614) |
| 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
载体简介
载体序列
LOCUS pFA6a-kanMX6-PGAL1-GFP 5432 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pFA6a-kanMX6-PGAL1-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5432)
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 5432)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5432
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter complement(1..19)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(2..19)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(100..119)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 219..241
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(279..297)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 365..469
/gene="bla"
/label=AmpR promoter
CDS 470..1330
/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(688..707)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1501..2089
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1990..2009
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2243..2260
/label=L4440
/note="L4440 vector, forward primer"
primer_bind 2347..2366
/label=T7
/note="T7 promoter, forward primer"
promoter 2347..2365
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
gene complement(2470..3826)
/label=kanMX
/note="yeast selectable marker conferring kanamycin
resistance (Wach et al., 1994)"
terminator 2470..2667
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
CDS complement(2673..3482)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 3396..3415
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
promoter complement(3483..3826)
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
promoter 3960..4401
/gene="S. cerevisiae GAL1"
/label=GAL1 promoter
/note="inducible promoter, regulated by Gal4"
protein_bind 3960..4077
/label=UAS
/bound_moiety="Gal4"
/note="upstream activating sequence mediating
Gal4-dependent induction"
primer_bind 4372..4395
/label=GAL1
/note="S. cerevisiae GAL1 promoter, forward primer"
CDS 4436..5149
/codon_start=1
/product="Aequoria victoria green fluorescent protein"
/label=GFP
/translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
ICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
VTAAGITHGMDELYK"
primer_bind complement(4463..4491)
/label=GFP-R
/note="GFP, reverse primer. Does NOT anneal to EGFP"
primer_bind 5086..5106
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
terminator 5170..5357
/gene="S. cerevisiae ADH1"
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
primer_bind complement(5225..5249)
/label=pHybLex-R
/note="ADH terminator, reverse primer"
ORIGIN
1 tctatagtgt cacctaaatc gtatgtgtat gatacataag gttatgtatt aattgtagcc
61 gcgttctaac gacaatatgt ccatatggtg cactctcagt acaatctgct ctgatgccgc
121 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
181 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
241 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
301 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
361 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
421 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
481 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
541 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
601 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
661 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
721 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
781 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
841 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
901 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
961 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
1021 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
1081 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
1141 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
1201 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
1261 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
1321 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
1381 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
1441 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
1501 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
1561 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
1621 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
1681 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
1741 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
1801 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
1861 ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg
1921 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
1981 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
2041 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
2101 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
2161 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
2221 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
2281 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaggttaa cctggcttat
2341 cgaaattaat acgactcact atagggagac cggcagatcc gcggccgcat aggccactag
2401 tggatctgat atcatcgatg aattcgagct cgtttaaact ggatggcggc gttagtatcg
2461 aatcgacagc agtatagcga ccagcattca catacgattg acgcatgata ttactttctg
2521 cgcacttaac ttcgcatctg ggcagatgat gtcgaggcga aaaaaaatat aaatcacgct
2581 aacatttgat taaaatagaa caactacaat ataaaaaaac tatacaaatg acaagttctt
2641 gaaaacaaga atctttttat tgtcagtact gattagaaaa actcatcgag catcaaatga
2701 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt
2761 aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct
2821 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
2881 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagctta
2941 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
3001 gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg
3061 ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
3121 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttg
3181 ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg
3241 gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca
3301 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
3361 aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
3421 aaatcagcat ccatgttgga atttaatcgc ggcctcgaaa cgtgagtctt ttccttaccc
3481 atggttgttt atgttcggat gtgatgtgag aactgtatcc tagcaagatt ttaaaaggaa
3541 gtatatgaaa gaagaacctc agtggcaaat cctaaccttt tatatttctc tacaggggcg
3601 cggcgtgggg acaattcaac gcgtctgtga ggggagcgtt tccctgctcg caggtctgca
3661 gcgaggagcc gtaatttttg cttcgcgccg tgcggccatc aaaatgtatg gatgcaaatg
3721 attatacatg gggatgtatg ggctaaatgt acgggcgaca gtcacatcat gcccctgagc
3781 tgcgcacgtc aagactgtca aggagggtat tctgggcctc catgtcgctg gccgggtgac
3841 ccggcgggga cgaggcaagc taaacagatc tgtaaagagc cccattatct tagcctaaaa
3901 aaaccttctc tttggaactt tcagtaatac gcttaactgc tcattgctat attgaagtac
3961 ggattagaag ccgccgagcg ggtgacagcc ctccgaagga agactctcct ccgtgcgtcc
4021 tcgtcttcac cggtcgcgtt cctgaaacgc agatgtgcct cgcgccgcac tgctccgaac
4081 aataaagatt ctacaatact agcttttatg gttatgaaga ggaaaaattg gcagtaacct
4141 ggccccacaa accttcaaat gaacgaatca aattaacaac cataggatga taatgcgatt
4201 agttttttag ccttatttct ggggtaatta atcagcgaag cgatgatttt tgatctatta
4261 acagatatat aaatgcaaaa actgcataac cactttaact aatactttca acattttcgg
4321 tttgtattac ttcttattca aatgtaataa aagtatcaac aaaaaattgt taatatacct
4381 ctatacttta acgtcaagga gaaaaaaccc ggatctcaaa atgtctttaa ttaacagtaa
4441 aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa
4501 tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac
4561 ccttaaattt atttgcacta ctggaaaact acctgttcca tggccaacac ttgtcactac
4621 tttcacttat ggtgttcaat gcttttcaag atacccagat catatgaaac ggcatgactt
4681 tttcaagagt gccatgcccg aaggttatgt acaggaaaga actatatttt tcaaagatga
4741 cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat
4801 cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt cttggacaca aattggaata
4861 caactataac tcacacaatg tatacatcat ggcagacaaa caaaagaatg gaatcaaagt
4921 taacttcaaa attagacaca acattgaaga tggaagcgtt caactagcag accattatca
4981 acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac
5041 acaatctgcc ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt
5101 tgtaacagct gctgggatta cacatggcat ggatgaacta tacaaatagg gcgcgccact
5161 tctaaataag cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa
5221 ataagtgtat acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg
5281 agtaactctt tcctgtaggt caggttgctt tctcaggtat agtatgaggt cgctcttatt
5341 gaccacacct ctaccggcag atccgctagg gataacaggg taatatagat ccgtcgacct
5401 gcagcgtacg aagcttcagc tggcggccgc gt
//
