| 质粒名称: | pFA6a-kanMX6 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 39296 |
| 存储实验室: | Jurg Bahler, 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: | 2300 bp |
| 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: | KanR,kanamycin resistance cassette |
| 物种来源: | -- |
| 插入大小: | 1600 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: |
A、 棉子型翻译延伸因子1a基因启动子（插入端N端）
A、 棉子型翻译延长因子1a基因终止子（插入端C端）
|
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BglII(未破坏) |
| 3' 克隆位点: | PmeI (未破坏) |
| 5' 测序引物 | Sp6_primer |
| 3' 测序引物 | T7_promoter |
|
文献引用方法 |
|
| 材料和方法部分: | pFA6a-kanMX6 was a gift from Jurg Bahler & John Pringle (Addgene plasmid # 39296 ; http://n2t.net/addgene:39296 ; RRID:Addgene_39296) |
| 参考文献部分: | 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
pFA6akanMX6载体基本信息
pFA6akanMX6载体图谱质粒图谱和多克隆位点信息
pFA6akanMX6载体简介
pFA6akanMX6载体序列
LOCUS pFA6a-kanMX6 3938 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pFA6a-kanMX6
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3938)
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 3938)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3938
/organism="synthetic DNA construct"
/mol_type="other DNA"
gene 115..1471
/label=kanMX
/note="yeast selectable marker conferring kanamycin
resistance (Wach et al., 1994)"
promoter 115..458
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
CDS 459..1268
/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(526..545)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
terminator 1274..1471
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
primer_bind complement(1575..1594)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1576..1594)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1681..1698)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1852..2440)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1932..1951)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2611..3471)
/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 3234..3253
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3472..3576)
/gene="bla"
/label=AmpR promoter
primer_bind 3644..3662
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(3700..3722)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 3822..3841
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 3922..2
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 3922..1
/label=SP6
/note="SP6 promoter, forward primer"
ORIGIN
1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
61 aggcgcgcca gatctgttta gcttgcctcg tccccgccgg gtcacccggc cagcgacatg
121 gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac
181 tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc catacatttt
241 gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga
301 aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata
361 aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta
421 ggatacagtt ctcacatcac atccgaacat aaacaaccat gggtaaggaa aagactcacg
481 tttcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
541 gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc
601 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
661 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
721 ctcctgatga tgcatggtta ctcaccactg cgatccccgg caaaacagca ttccaggtat
781 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
841 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
901 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
961 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac
1021 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
1081 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
1141 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
1201 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
1261 ttttctaatc agtactgaca ataaaaagat tcttgttttc aagaacttgt catttgtata
1321 gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt atattttttt
1381 tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt
1441 caatcgtatg tgaatgctgg tcgctatact gctgtcgatt cgatactaac gccgccatcc
1501 agtttaaacg agctcgaatt catcgatgat atcagatcca ctagtggcct atgcggccgc
1561 ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg ttaacctgca
1621 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
1681 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
1741 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
1801 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
1861 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
1921 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
1981 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
2041 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
2101 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
2161 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
2221 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
2281 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
2341 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
2401 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
2461 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
2521 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
2581 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
2641 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
2701 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
2761 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
2821 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
2881 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
2941 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
3001 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
3061 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
3121 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
3181 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
3241 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
3301 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
3361 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
3421 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
3481 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
3541 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
3601 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
3661 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
3721 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
3781 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
3841 actgagagtg caccatatgg acatattgtc gttagaacgc ggctacaatt aatacataac
3901 cttatgtatc atacacatac gatttaggtg acactata
//
