| 出品公司: | Addgene |
|---|---|
| 载体名称: | pFA6a-GFP(S65T)-KanMX6 |
| 质粒类型: | 酿酒酵母表达载体 |
| 存储人: | Jurg Bahler, John Pringle |
| 高拷贝/低拷贝: | 高靠背 |
| 启动子: | SP6 |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 4882bp |
| 5' 测序引物及序列: | Sp6-F: ATTTAGGTGACACTATAG |
| 3' 测序引物及序列: | T7: TAATACGACTCACTATAGGG |
| 载体标签: | N-GFP (S65T) |
| 载体抗性: | 氨苄 |
| 筛选标记: | 卡那霉素 |
| 备注: | -- |
| 产品目录号: | 39292 |
| 稳定性: | 稳定 |
| 病毒/非病毒: | 非病毒 |
| 质粒引用： | pFA6a-GFP(S65T)-kanMX6 was a gift from Jurg Bahler & John Pringle (Addgene plasmid # 39292 ; http://n2t.net/addgene:39292 ; RRID:Addgene_39292) |
| 参考文献： | Heterologous modules for efficient and versatile PCR-based gene targeting in Schizosaccharomyces pombe. Bahler J, Wu JQ, Longtine MS, Shah NG, McKenzie A 3rd, Steever AB, Wach A, Philippsen P, Pringle JR. Yeast. 1998 Jul;14(10):943-51. 10.1002/(SICI)1097-0061(199807)14:10<943::AID-YEA292>3.0.CO;2-Y PubMed 9717240 |
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
-kanMX6载体图谱.png)
载体简介
载体序列
LOCUS pFA6a-GFP(S65T)-kanMX6 4882 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pFA6a-GFP(S65T)-kanMX6
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4882)
AUTHORS Bahler J, Wu JQ, Longtine MS, Shah NG, McKenzie A 3rd, Steever AB,
Wach A, Philippsen P, Pringle JR
TITLE Heterologous modules for efficient and versatile PCR-based gene
targeting in Schizosaccharomyces pombe.
JOURNAL Yeast. 1998 Jul;14(10):943-51.
PUBMED 9717240
REFERENCE 2 (bases 1 to 4882)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4882
/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"
gene 1057..2413
/label=kanMX
/note="yeast selectable marker conferring kanamycin
resistance (Wach et al., 1994)"
promoter 1057..1400
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
CDS 1401..2210
/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 complement(1468..1487)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
terminator 2216..2413
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
primer_bind complement(2517..2536)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(2518..2536)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2623..2640)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2794..3382)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2874..2893)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3553..4413)
/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 4176..4195
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4414..4518)
/gene="bla"
/label=AmpR promoter
primer_bind 4586..4604
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(4642..4664)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 4764..4783
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 4864..4882
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 4864..4881
/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 atgggtaagg aaaagactca cgtttcgagg ccgcgattaa
1441 attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat
1501 caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg tttctgaaac
1561 atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga
1621 cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt
1681 tactcaccac tgcgatcccc ggcaaaacag cattccaggt attagaagaa tatcctgatt
1741 caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg
1801 tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa
1861 tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg
1921 aacaagtctg gaaagaaatg cataagcttt tgccattctc accggattca gtcgtcactc
1981 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg
2041 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc
2101 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc
2161 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa tcagtactga
2221 caataaaaag attcttgttt tcaagaactt gtcatttgta tagttttttt atattgtagt
2281 tgttctattt taatcaaatg ttagcgtgat ttatattttt tttcgcctcg acatcatctg
2341 cccagatgcg aagttaagtg cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct
2401 ggtcgctata ctgctgtcga ttcgatacta acgccgccat ccagtttaaa cgagctcgaa
2461 ttcatcgatg atatcagatc cactagtggc ctatgcggcc gcggatctgc cggtctccct
2521 atagtgagtc gtattaattt cgataagcca ggttaacctg cattaatgaa tcggccaacg
2581 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
2641 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
2701 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
2761 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
2821 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
2881 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
2941 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
3001 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
3061 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
3121 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
3181 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
3241 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
3301 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
3361 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
3421 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
3481 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
3541 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
3601 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
3661 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
3721 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
3781 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
3841 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
3901 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
3961 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
4021 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
4081 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
4141 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
4201 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
4261 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
4321 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
4381 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
4441 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
4501 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
4561 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
4621 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
4681 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
4741 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
4801 ggacatattg tcgttagaac gcggctacaa ttaatacata accttatgta tcatacacat
4861 acgatttagg tgacactata ga
//
