| 质粒名称: | pUDE731 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 103008 |
| 存储实验室: | Jean-Marc Daran |
|
载体骨架基本信息 |
|
| 载体名称: | pMEL10 |
| 载体出品公司: | Mans, R. et al (2015) CRISPR/Cas9: a molecular Swiss army knife FEMS Yeast Res 15 |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5658 bp |
| 完整质粒大小: | 9695 bp |
| 载体类型: | 酵母表达 |
| 筛选标记: | URA3 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Fncpf1 |
| 物种来源: | Francisella tularensis subsp. novicida U112 |
| 插入大小: | 4038 bp |
| 基因突变: | human codon optimized |
| GenBank编号 | CP000439.1 CP000439.1 |
| 启动子 | TEF1 |
| 标签/融合蛋白: |
NLS（插接件上的C端子）
3HA（插接件上的C端子）
|
| 克隆方法: | Gibson克隆 |
| 5' 测序引物 | GCCTACGGTTCCCGAAGTATGCTGCTGATGTCTGGCTATACCTATCCGTCTACGTGAATACCCTCACTAAAGGGAACAAAAG |
| 3' 测序引物 | GTTGAACATTCTTAGGCTGGTCGAATCATTTAGACACGGGCATCGTCCTCTCGAAAGGTGGGCCGCAAATTAAAGCCTTCGAG |
|
文献引用方法 |
|
| 材料和方法部分: | pUDE731 was a gift from Jean-Marc Daran (Addgene plasmid # 103008 ; http://n2t.net/addgene:103008 ; RRID:Addgene_103008) |
| 参考文献部分: | FnCpf1, a novel and efficient genome editing tool for Saccharomyces cerevisiae. Świat MA, Dashko S, den Ridder M, Wijsman M, van der Oost J, Daran JM Daran-Lapujade P.. Nucleic Acids Research (2017), gkx1007 10.1093/nar/gkx1007 |
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 pUDE731 9589 bp ds-DNA circular SYN 23-OCT-2019
DEFINITION S. cerevisiae Episomal plasmid harboring Fncpf1 under the control of
TDH3p.
ACCESSION .
VERSION .
KEYWORDS pUDE731
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9589)
AUTHORS Swiat MA, Dashko S, den Ridder M, Wijsman M, van der Oost J, Daran
JM Daran-Lapujade P.
TITLE FnCpf1, a novel and efficient genome editing tool for Saccharomyces
cerevisiae.
JOURNAL Nucleic Acids Research (2017), gkx1007
REFERENCE 2 (bases 1 to 9589)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Oct 23, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9589
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 99..146
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
rep_origin 127..1109
/label=2u ori
/note="yeast 2u plasmid origin of replication"
promoter 1136..1240
/gene="bla"
/label=AmpR promoter
CDS 1241..2101
/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(1459..1478)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2272..2860
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2761..2780
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3014..3031
/label=L4440
/note="L4440 vector, forward primer"
promoter 3158..3556
/gene="S. cerevisiae TEF1"
/label=TEF1 promoter
/note="promoter for EF-1-alpha"
CDS 7461..7508
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 7515..7595
/codon_start=1
/product="three tandem HA epitope tags"
/label=3xHA
/translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
primer_bind complement(7608..7626)
/label=CYC1
/note="CYC1 transcription termination signal, reverse
primer"
terminator 7638..7827
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
protein_bind 7981..8014
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
protein_bind 9466..9499
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
ORIGIN
1 tacaggcaac acgcagatat aggtgcgacg tgaacagtga gctgtatgtg cgcagctcgc
61 gttgcatttt cggaagcgct cgttttcgga aacgctttga agttcctatt ccgaagttcc
121 tattctctag aaagtatagg aacttcagag cgcttttgaa aaccaaaagc gctctgaaga
181 cgcactttca aaaaaccaaa aacgcaccgg actgtaacga gctactaaaa tattgcgaat
241 accgcttcca caaacattgc tcaaaagtat ctctttgcta tatatctctg tgctatatcc
301 ctatataacc tacccatcca cctttcgctc cttgaacttg catctaaact cgacctctac
361 attttttatg tttatctcta gtattactct ttagacaaaa aaattgtagt aagaactatt
421 catagagtga atcgaaaaca atacgaaaat gtaaacattt cctatacgta gtatatagag
481 acaaaataga agaaaccgtt cataattttc tgaccaatga agaatcatca acgctatcac
541 tttctgttca caaagtatgc gcaatccaca tcggtataga atataatcgg ggatgccttt
601 atcttgaaaa aatgcacccg cagcttcgct agtaatcagt aaacgcggga agtggagtca
661 ggcttttttt atggaagaga aaatagacac caaagtagcc ttcttctaac cttaacggac
721 ctacagtgca aaaagttatc aagagactgc attatagagc gcacaaagga gaaaaaaagt
781 aatctaagat gctttgttag aaaaatagcg ctctcgggat gcatttttgt agaacaaaaa
841 agaagtatag attctttgtt ggtaaaatag cgctctcgcg ttgcatttct gttctgtaaa
901 aatgcagctc agattctttg tttgaaaaat tagcgctctc gcgttgcatt tttgttttac
961 aaaaatgaag cacagattct tcgttggtaa aatagcgctt tcgcgttgca tttctgttct
1021 gtaaaaatgc agctcagatt ctttgtttga aaaattagcg ctctcgcgtt gcatttttgt
1081 tctacaaaat gaagcacaga tgcttcgttc aggtggcact tttcggggaa atgtgcgcgg
1141 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
1201 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
1261 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
1321 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
1381 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
1441 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
1501 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
1561 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
1621 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
1681 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
1741 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
1801 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
1861 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
1921 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
1981 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
2041 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
2101 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
2161 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
2221 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
2281 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
2341 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
2401 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
2461 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
2521 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
2581 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
2641 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
2701 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
2761 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
2821 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
2881 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
2941 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
3001 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
3061 gcctctcgcc tacggttccc gaagtatgct gctgatgtct ggctatacct atccgtctac
3121 gtgaataccc tcactaaagg gaacaaaagc tggagctcat agcttcaaaa tgtttctact
3181 ccttttttac tcttccagat tttctcggac tccgcgcatc gccgtaccac ttcaaaacac
3241 ccaagcacag catactaaat ttcccctctt tcttcctcta gggtgtcgtt aattacccgt
3301 actaaaggtt tggaaaagaa aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg
3361 caataaaaat ttttatcacg tttctttttc ttgaaaattt tttttttgat ttttttctct
3421 ttcgatgacc tcccattgat atttaagtta ataaacggtc ttcaatttct caagtttcag
3481 tttcattttt cttgttctat tacaactttt tttacttctt gctcattaga aagaaagcat
3541 agcaatctaa tctaagtttt atgagcatct accaggagtt cgtcaacaag tattcactga
3601 gtaagacact gcggttcgag ctgatcccac agggcaagac actggagaac atcaaggccc
3661 gaggcctgat tctggacgat gagaagcggg caaaagacta taagaaagcc aagcagatca
3721 ttgataaata ccaccagttc tttatcgagg aaattctgag ctccgtgtgc atcagtgagg
3781 atctgctgca gaattactca gacgtgtact tcaagctgaa gaagagcgac gatgacaacc
3841 tgcagaagga cttcaagtcc gccaaggaca ccatcaagaa acagattagc gagtacatca
3901 aggactccga aaagtttaaa aatctgttca accagaatct gatcgatgct aagaaaggcc
3961 aggagtccga cctgatcctg tggctgaaac agtctaagga caatgggatt gaactgttca
4021 aggctaactc cgatatcact gatattgacg aggcactgga aatcatcaag agcttcaagg
4081 gatggaccac atactttaaa ggcttccacg agaaccgcaa gaacgtgtac tccagcaacg
4141 acattcctac ctccatcatc taccgaatcg tcgatgacaa tctgccaaag ttcctggaga
4201 acaaggccaa atatgaatct ctgaaggaca aagctcccga ggcaattaat tacgaacaga
4261 tcaagaaaga tctggctgag gaactgacat tcgatatcga ctataagact agcgaggtga
4321 accagagggt cttttccctg gacgaggtgt ttgaaatcgc caatttcaac aattacctga
4381 accagtccgg cattactaaa ttcaatacca tcattggcgg gaagtttgtg aacggggaga
4441 ataccaagcg caagggaatt aacgaataca tcaatctgta tagccagcag atcaacgaca
4501 aaactctgaa gaaatacaag atgtctgtgc tgttcaaaca gatcctgagt gataccgagt
4561 ccaagtcttt tgtcattgat aaactggaag atgactcaga cgtggtcact accatgcaga
4621 gcttttatga gcagatcgcc gctttcaaga cagtggagga aaaatctatt aaggaaactc
4681 tgagtctgct gttcgatgac ctgaaagccc agaagctgga cctgagtaag atctacttca
4741 aaaacgataa gagtctgaca gacctgtcac agcaggtgtt tgatgactat tccgtgattg
4801 ggaccgccgt cctggagtac attacacagc agatcgctcc aaagaacctg gataatccct
4861 ctaagaaaga gcaggaactg atcgctaaga aaaccgagaa ggcaaaatat ctgagtctgg
4921 aaacaattaa gctggcactg gaggagttca acaagcacag ggatattgac aaacagtgcc
4981 gctttgagga aatcctggcc aacttcgcag ccatccccat gatttttgat gagatcgccc
5041 agaacaaaga caatctggct cagatcagta ttaagtacca gaaccagggc aagaaagacc
5101 tgctgcaggc ttcagcagaa gatgacgtga aagccatcaa ggatctgctg gaccagacca
5161 acaatctgct gcacaagctg aaaatcttcc atattagtca gtcagaggat aaggctaata
5221 tcctggataa agacgaacac ttctacctgg tgttcgagga atgttacttc gagctggcaa
5281 acattgtccc cctgtataac aagattagga actacatcac acagaagcct tactctgacg
5341 agaagtttaa actgaacttc gaaaatagta ccctggccaa cgggtgggat aagaacaagg
5401 agcctgacaa cacagctatc ctgttcatca aggatgacaa gtactatctg ggagtgatga
5461 ataagaaaaa caataagatc ttcgatgaca aagccattaa ggagaacaaa ggggaaggat
5521 acaagaaaat cgtgtataag ctgctgcccg gcgcaaataa gatgctgcct aaggtgttct
5581 tcagcgccaa gagtatcaaa ttctacaacc catccgagga catcctgcgg attagaaatc
5641 actcaacaca tactaagaac gggagccccc agaagggata tgagaaattt gagttcaaca
5701 tcgaggattg caggaagttt attgacttct acaagcagag catctccaaa caccctgaat
5761 ggaaggattt tggcttccgg ttttccgaca cacagagata taactctatc gacgagttct
5821 accgcgaggt ggaaaatcag gggtataagc tgacttttga gaacatttct gaaagttaca
5881 tcgacagcgt ggtcaatcag ggaaagctgt acctgttcca gatctataac aaagattttt
5941 cagcatacag caagggcaga ccaaacctgc atacactgta ctggaaggcc ctgttcgatg
6001 agaggaatct gcaggacgtg gtctataaac tgaacggaga ggccgaactg ttttaccgga
6061 agcagtctat tcctaagaaa atcactcacc cagctaagga ggccatcgct aacaagaaca
6121 aggacaatcc taagaaagag agcgtgttcg aatacgatct gattaaggac aagcggttca
6181 ccgaagataa gttctttttc cattgtccaa tcaccattaa cttcaagtca agcggcgcta
6241 acaagttcaa cgacgagatc aatctgctgc tgaaggaaaa agcaaacgat gtgcacatcc
6301 tgagcattga ccgaggagag cggcatctgg cctactatac cctggtggat ggcaaaggga
6361 atatcattaa gcaggataca ttcaacatca ttggcaatga ccggatgaaa accaactacc
6421 acgataaact ggctgcaatc gagaaggata gagactcagc taggaaggac tggaagaaaa
6481 tcaacaacat taaggagatg aaggaaggct atctgagcca ggtggtccat gagattgcaa
6541 agctggtcat cgaatacaat gccattgtgg tgttcgagga tctgaacttc ggctttaaga
6601 gggggcgctt taaggtggaa aaacaggtct atcagaagct ggagaaaatg ctgatcgaaa
6661 agctgaatta cctggtgttt aaagataacg agttcgacaa gaccggaggc gtcctgagag
6721 cctaccagct gacagctccc tttgaaactt tcaagaaaat gggaaaacag acaggcatca
6781 tctactatgt gccagccgga ttcacttcca agatctgccc cgtgaccggc tttgtcaacc
6841 agctgtaccc taaatatgag tcagtgagca agtcccagga atttttcagc aagttcgata
6901 agatctgtta taatctggac aaggggtact tcgagttttc cttcgattac aagaacttcg
6961 gcgacaaggc cgctaagggg aaatggacca ttgcctcctt cggatctcgc ctgatcaact
7021 ttcgaaattc cgataaaaac cacaattggg acactaggga ggtgtaccca accaaggagc
7081 tggaaaagct gctgaaagac tactctatcg agtatggaca tggcgaatgc atcaaggcag
7141 ccatctgtgg cgagagtgat aagaaatttt tcgccaagct gacctcagtg ctgaatacaa
7201 tcctgcagat gcggaactca aagaccggga cagaactgga ctatctgatt agccccgtgg
7261 ctgatgtcaa cggaaacttc ttcgacagca gacaggcacc caaaaatatg cctcaggatg
7321 cagacgccaa cggggcctac cacatcgggc tgaagggact gatgctgctg ggccggatca
7381 agaacaatca ggaggggaag aagctgaacc tggtcattaa gaacgaggaa tacttcgagt
7441 ttgtccagaa tagaaataac aaaaggccgg cggccacgaa aaaggccggc caggcaaaaa
7501 agaaaaaggg atcctaccca tacgatgttc cagattacgc ttatccctac gacgtgcctg
7561 attatgcata cccatatgat gtccccgact atgcctaaat tagttatgtc acgcttacat
7621 tcacgccctc cccccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc
7681 taggtcccta tttatttttt tatagttatg ttagtattaa gaacgttatt tatatttcaa
7741 atttttcttt tttttctgta cagacgcgtg tacgcatgta acattatact gaaaaccttg
7801 cttgagaagg ttttgggacg ctcgaaggct ttaatttgcg gcccaccttt cgagaggacg
7861 atgcccgtgt ctaaatgatt cgaccagcct aagaatgttc aacggctggt cgaatcattt
7921 agacacgggc atcgtcctct cgaaaggtgg ctaggccact agtggatctg atatcaccta
7981 ataacttcgt atagcataca ttatacgaag ttatattaag ggttctcgag agctcgtttt
8041 atttaggttc tatcgaggag aaaaagcgac aagaagagat agaccatgga taaatgatta
8101 tgttctaaac actcctcaga agctcatcga actgtcatcc tgcgtgaaga ttaaaatcca
8161 acttagaaat ttcgagctta cggagacaat catatgggag aagcaattgg aagatagaaa
8221 aaaggtactc ggtacataaa tatatgtgat tctgggtaga agatcggtct gcattggatg
8281 gtggtaacgc atttttttac acacattact tgcctcgagc atcaaatggt ggttattcgt
8341 ggatctatat cacgtgattt gcttaagaat tgtcgttcat ggtgacactt ttagctttga
8401 catgattaag ctcatctcaa ttgatgttat ctaaagtcat ttcaactatc taagatgtgg
8461 ttgtgattgg gccattttgt gaaagccagt acgccagcgt caatacactc ccgtcaatta
8521 gttgcaccat gtccacaaaa tcatatacca gtagagctga gactcatgca agtccggttg
8581 catcgaaact tttacgttta atggatgaaa agaagaccaa tttgtgtgct tctcttgacg
8641 ttcgttcgac tgatgagcta ttgaaacttg ttgaaacgtt gggtccatac atttgccttt
8701 tgaaaacaca cgttgatatc ttggatgatt tcagttatga gggtactgtc gttccattga
8761 aagcattggc agagaaatac aagttcttga tatttgagga cagaaaattc gccgatatcg
8821 gtaacacagt caaattacaa tatacatcgg gcgtttaccg tatcgcagaa tggtctgata
8881 tcaccaacgc ccacggggtt actggtgctg gtattgttgc tggcttgaaa caaggtgcgc
8941 aagaggtcac caaagaacca aggggattat tgatgcttgc tgaattgtct tccaagggtt
9001 ctctagcaca cggtgaatat actaagggta ccgttgatat tgcaaagagt gataaagatt
9061 tcgttattgg gttcattgct cagaacgata tgggaggaag agaagaaggg tttgattggc
9121 taatcatgac cccaggtgta ggtttagacg acaaaggcga tgcattgggt cagcagtaca
9181 gaaccgtcga cgaagttgta agtggtggat cagatatcat cattgttggc agaggacttt
9241 tcgccaaggg tagagatcct aaggttgaag gtgaaagata cagaaatgct ggatgggaag
9301 cgtaccaaaa gagaatcagc gctccccatt aattatacag gaaacttaat agaacaaatc
9361 acatatttaa tctaatagcc acctgcattg gcacggtgca acactacttc aacttcatct
9421 tacaaaaaga tcacgtgatc tgttgtattg ggatctctag acctaataac ttcgtatagc
9481 atacattata cgaagttata ttaagggttg tcgacctgca gcgtacgaag gtgcctattg
9541 atgatctggc ggaatgtctg ccgtgccata gccatgcctt cacatatag
//
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