| 质粒名称: | pFA6a-kanMX-tetO-Pcyc1-GFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 41025 |
| 存储实验室: | Michael Nick Boddy |
|
载体骨架基本信息 |
|
| 载体名称: | pFA6a |
| 完整质粒大小: | 5395 bp |
| 载体类型: | N-terminal tagging/promoter replacement |
| 筛选标记: | G418 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | tetO7-Pcyc1-GFP |
|
文献引用方法 |
|
| 材料和方法部分: | pFA6a-kanMX-tetO-Pcyc1-GFP was a gift from Michael Nick Boddy (Addgene plasmid # 41025 ; http://n2t.net/addgene |
| 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 |
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-kanMX-tetO-Pcyc1-GFP 5395 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pFA6a-kanMX-tetO-Pcyc1-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5395)
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 5395)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5395
/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"
protein_bind 3909..4179
/label=tetracycline response element
/bound_moiety="tetracycline repressor TetR"
/note="contains seven copies of the tetracycline operator
tetO"
CDS 4399..5112
/codon_start=1
/product="Aequoria victoria green fluorescent protein"
/label=GFP
/translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
ICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
VTAAGITHGMDELYK"
primer_bind complement(4426..4454)
/label=GFP-R
/note="GFP, reverse primer. Does NOT anneal to EGFP"
primer_bind 5049..5069
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
terminator 5133..5320
/gene="S. cerevisiae ADH1"
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
primer_bind complement(5188..5212)
/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 tgggtaatat agatcaattc ctcgatcgag
3901 tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag
3961 tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa
4021 gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc
4081 tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa
4141 gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagctcggt
4201 accctatggc atgcatgtgc tctgtatgta tataaaactc ttgttttctt cttttctcta
4261 aatattcttt ccttatacat taggtccttt gtagcataaa ttactatact tctatagaca
4321 cgcaaacaca aatacacaca ctaaattacc ggatcaattc gggggatccg tttatcatcc
4381 tggaggatgt taattaacag taaaggagaa gaacttttca ctggagttgt cccaattctt
4441 gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga gggtgaaggt
4501 gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa actacctgtt
4561 ccatggccaa cacttgtcac tactttcact tatggtgttc aatgcttttc aagataccca
4621 gatcatatga aacggcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa
4681 agaactatat ttttcaaaga tgacgggaac tacaagacac gtgctgaagt caagtttgaa
4741 ggtgataccc ttgttaatag aatcgagtta aaaggtattg attttaaaga agatggaaac
4801 attcttggac acaaattgga atacaactat aactcacaca atgtatacat catggcagac
4861 aaacaaaaga atggaatcaa agttaacttc aaaattagac acaacattga agatggaagc
4921 gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta
4981 ccagacaacc attacctgtc cacacaatct gccctttcga aagatcccaa cgaaaagaga
5041 gaccacatgg tccttcttga gtttgtaaca gctgctggga ttacacatgg catggatgaa
5101 ctatacaaat agggcgcgcc acttctaaat aagcgaattt cttatgattt atgattttta
5161 ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac tcttaggttt
5221 taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg ctttctcagg
5281 tatagtatga ggtcgctctt attgaccaca cctctaccgg cagatccgct agggataaca
5341 gggtaatata gatccgtcga cctgcagcgt acgaagcttc agctggcggc cgcgt
//
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