| 质粒名称: | pFA6a-hphMX-tetO-Pcyc1-3xFLAG |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 41020 |
| 存储实验室: | Michael Nick Boddy |
|
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: | N-末端标记/启动子替换 |
| 筛选标记: | 潮霉素 |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | tetO7-Pcyc1-3xFLAG |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pFA6a-hphMX-tetO-Pcyc1-3xFLAG was a gift from Michael Nick Boddy (Addgene plasmid # 41020 ; http://n2t.net/addgene:41020 ; RRID:Addgene_41020) |
| 参考文献部分: | A new versatile system for rapid control of gene expression in the fission yeast Schizosaccharomyces pombe. Zilio N, Wehrkamp-Richter S, Boddy MN. Yeast. 2012 Aug 29. doi: 10.1002/yea.2920. 10.1002/yea.2920 PubMed 22968950 |
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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-hphMX-tetO-Pcyc1-3xFLAG 4975 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pFA6a-hphMX-tetO-Pcyc1-3xFLAG
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4975)
AUTHORS Zilio N, Wehrkamp-Richter S, Boddy MN
TITLE A new versatile system for rapid control of gene expression in the
fission yeast Schizosaccharomyces pombe.
JOURNAL Yeast. 2012 Aug 29. doi: 10.1002/yea.2920.
PUBMED 22968950
REFERENCE 2 (bases 1 to 4975)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4975
/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..4045)
/label=hphMX6
/note="yeast selectable marker conferring hygromycin
resistance"
terminator 2470..2667
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
CDS complement(2673..3701)
/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"
promoter complement(3702..4045)
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
protein_bind 4128..4398
/label=tetracycline response element
/bound_moiety="tetracycline repressor TetR"
/note="contains seven copies of the tetracycline operator
tetO"
CDS 4618..4689
/codon_start=1
/product="three tandem FLAG(R) epitope tags"
/label=3xFLAG
/translation="DYKDDDDKDYKDDDDKDYKDDDDK"
terminator 4713..4900
/gene="S. cerevisiae ADH1"
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
primer_bind complement(4768..4792)
/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 gattattcct ttgccctcgg acgagtgctg
2701 gggcgtcggt ttccactatc ggcgagtact tctacacagc catcggtcca gacggccgcg
2761 cttctgcggg cgatttgtgt acgcccgaca gtcccggctc cggatcggac gattgcgtcg
2821 catcgaccct gcgcccaagc tgcatcatcg aaattgccgt caaccaagct ctgatagagt
2881 tggtcaagac caatgcggag catatacgcc cggagccgcg gcgatcctgc aagctccgga
2941 tgcctccgct cgaagtagcg cgtctgctgc tccatacaag ccaaccacgg cctccagaag
3001 aagatgttgg cgacctcgta ttgggaatcc ccgaacatcg cctcgctcca gtcaatgacc
3061 gctgttatgc ggccattgtc cgtcaggaca ttgttggagc cgaaatccgc gtgcacgagg
3121 tgccggactt cggggcagtc ctcggcccaa agcatcagct catcgagagc ctgcgcgacg
3181 gacgcactga cggtgtcgtc catcacagtt tgccagtgat acacatgggg atcagcaatc
3241 gcgcatatga aatcacgcca tgtagtgtat tgaccgattc cttgcggtcc gaatgggccg
3301 aacccgctcg tctggctaag atcggccgca gcgatcgcat ccatggcctc cgcgaccggc
3361 tgcagaacag cgggcagttc ggtttcaggc aggtcttgca acgtgacacc ctgtgcacgg
3421 cgggagatgc aataggtcag gctctcgctg aattccccaa tgtcaagcac ttccggaatc
3481 gggagcgcgg ccgatgcaaa gtgccgataa acataacgat ctttgtagaa accatcggcg
3541 cagctattta cccgcaggac atatccacgc cctcctacat cgaagctgaa agcacgagat
3601 tcttcgccct ccgagagctg catcaggtcg gagacgctgt cgaacttttc gatcagaaac
3661 ttctcgacag acgtcgcggt gagttcaggc tttttaccca tggttgttta tgttcggatg
3721 tgatgtgaga actgtatcct agcaagattt taaaaggaag tatatgaaag aagaacctca
3781 gtggcaaatc ctaacctttt atatttctct acaggggcgc ggcgtgggga caattcaacg
3841 cgtctgtgag gggagcgttt ccctgctcgc aggtctgcag cgaggagccg taatttttgc
3901 ttcgcgccgt gcggccatca aaatgtatgg atgcaaatga ttatacatgg ggatgtatgg
3961 gctaaatgta cgggcgacag tcacatcatg cccctgagct gcgcacgtca agactgtcaa
4021 ggagggtatt ctgggcctcc atgtcgctgg ccgggtgacc cggcggggac gaggcaagct
4081 aaacagatct gggtaatata gatcaattcc tcgatcgagt ttaccactcc ctatcagtga
4141 tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa agtgaaagtc
4201 gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat
4261 cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg
4321 aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact
4381 ccctatcagt gatagagaaa agtgaaagtc gagctcggta ccctatggca tgcatgtgct
4441 ctgtatgtat ataaaactct tgttttcttc ttttctctaa atattctttc cttatacatt
4501 aggtcctttg tagcataaat tactatactt ctatagacac gcaaacacaa atacacacac
4561 taaattaccg gatcaattcg ggggatccgt ttatcatcct ggaggttaat taatatggac
4621 tacaaggacg acgatgacaa ggattacaaa gatgacgacg ataaggacta taaggacgat
4681 gatgataaat agggcgcgcc acttctaaat aagcgaattt cttatgattt atgattttta
4741 ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac tcttaggttt
4801 taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg ctttctcagg
4861 tatagtatga ggtcgctctt attgaccaca cctctaccgg cagatccgct agggataaca
4921 gggtaatata gatccgtcga cctgcagcgt acgaagcttc agctggcggc cgcgt
//
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