| 质粒名称: | pFA6a-5FLAG-hphMX6 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 19342 |
| 存储实验室: | Eishi Noguchi |
|
载体骨架基本信息 |
|
| 载体名称: | pFA6a-5FLAG-kanMX6 |
| 载体出品公司: | -- |
| 空载体大小: | 2900 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 酵母表达 |
| 筛选标记: | 潮霉素 |
| 载体抗性: | 氨苄青霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | hphMX6 |
| 物种来源: | E. coli |
| 插入大小: | 1700 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BglII（如果已销毁，则未知） |
| 3’克隆位点: | PmeI（如果已销毁，则未知） |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pFA6a-5FLAG-hphMX6 was a gift from Eishi Noguchi (Addgene plasmid # 19342 ; http://n2t.net/addgene:19342 ; RRID:Addgene_19342) |
| 参考文献部分: | A vector system for genomic FLAG epitope-tagging in Schizosaccharomyces pombe. Noguchi C, Garabedian MV, Malik M, Noguchi E. Biotechnol J. 2008 Aug 26. ():. 10.1002/biot.200800140 PubMed 18729046 |
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-5FLAG-hphMX6 4531 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pFA6a-5FLAG-hphMX6
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4531)
AUTHORS Noguchi C, Garabedian MV, Malik M, Noguchi E
TITLE A vector system for genomic FLAG epitope-tagging in
Schizosaccharomyces pombe.
JOURNAL Biotechnol J. 2008 Aug 26. ():.
PUBMED 18729046
REFERENCE 2 (bases 1 to 4531)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4531
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 69..200
/codon_start=1
/product="five tandem FLAG(R) epitope tags"
/label=5xFLAG
/translation="DYKDDDDKDYKDDDDKLMDYKDDDDKDYKDDDDKLMDYKDDDDK"
terminator 227..414
/gene="S. cerevisiae ADH1"
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
primer_bind complement(282..306)
/label=pHybLex-R
/note="ADH terminator, reverse primer"
gene 489..2064
/label=hphMX6
/note="yeast selectable marker conferring hygromycin
resistance"
promoter 489..832
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
CDS 833..1861
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
KE"
terminator 1867..2064
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
primer_bind complement(2168..2187)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(2169..2187)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2274..2291)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2445..3033)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2525..2544)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3204..4064)
/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 3827..3846
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4065..4169)
/gene="bla"
/label=AmpR promoter
primer_bind 4237..4255
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(4293..4315)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 4415..4434
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 4515..2
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 4515..1
/label=SP6
/note="SP6 promoter, forward primer"
ORIGIN
1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
61 atcatatgga ctacaaggac gacgatgaca aggattacaa agatgacgac gataagctta
121 tggactacaa ggacgacgat gacaaggatt acaaagatga cgacgataag cttatggact
181 acaaggacga cgatgacaag catatgggcg cgccacttct aaataagcga atttcttatg
241 atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca aattttaaag
301 tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc tgtaggtcag
361 gttgctttct caggtatagt atgaggtcgc tcttattgac cacacctcta ccggcagatc
421 cgctagggat aacagggtaa tatagatctg tttagcttgc ctcgtccccg ccgggtcacc
481 cggccagcga catggaggcc cagaataccc tccttgacag tcttgacgtg cgcagctcag
541 gggcatgatg tgactgtcgc ccgtacattt agcccataca tccccatgta taatcatttg
601 catccataca ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga
661 cctgcgagca gggaaacgct cccctcacag acgcgttgaa ttgtccccac gccgcgcccc
721 tgtagagaaa tataaaaggt taggatttgc cactgaggtt cttctttcat atacttcctt
781 ttaaaatctt gctaggatac agttctcaca tcacatccga acataaacaa ccatgggtaa
841 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
901 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
961 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
1021 tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga
1081 attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga
1141 cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat
1201 cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg
1261 tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg
1321 gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat
1381 gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa
1441 caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt
1501 cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat
1561 ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct
1621 ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa
1681 tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg
1741 gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt
1801 agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata
1861 atcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt
1921 tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc
1981 gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt
2041 atgtgaatgc tggtcgctat actgctgtcg attcgatact aacgccgcca tccagtttaa
2101 acgagctcga attcatcgat gatatcagat ccactagtgg cctatgcggc cgcggatctg
2161 ccggtctccc tatagtgagt cgtattaatt tcgataagcc aggttaacct gcattaatga
2221 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
2281 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
2341 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
2401 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
2461 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
2521 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
2581 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
2641 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
2701 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
2761 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
2821 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
2881 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
2941 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
3001 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
3061 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
3121 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
3181 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
3241 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
3301 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
3361 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
3421 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
3481 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
3541 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
3601 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
3661 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
3721 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
3781 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
3841 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
3901 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
3961 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
4021 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
4081 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
4141 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
4201 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
4261 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
4321 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
4381 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
4441 gtgcaccata tggacatatt gtcgttagaa cgcggctaca attaatacat aaccttatgt
4501 atcatacaca tacgatttag gtgacactat a
//
