| 质粒名称: | pYES2-gRNA-hyg-MCS |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 107734 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pYES2 |
| 空载体大小: | 5856 bp |
| 完整质粒大小: | 6244 bp |
| 载体类型: | Yeast Expression, CRISPR |
| 筛选标记: | Hygromycin |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | X-3 site gRNA expression cassettes / HXK1 /YFR053C |
| 物种来源: | S. cerevisiae (budding yeast) |
| GenBank编号 | 850614 |
| 启动子: | SNR52p |
| 克隆方法: | 未知 |
| 5’测序引物 | NA |
|
文献引用方法 |
|
| 材料和方法部分: |
pYES2-gRNA-hyg-MCS was a gift from Sheng Yang (Addgene plasmid # 107734 ; http://n2t.net/addgene:107734 ; RRID:Addgene_107734)
|
| 参考文献部分: | Unraveling the genetic basis of fast l-arabinose consumption on top of recombinant xylose-fermenting Saccharomyces cerevisiae. Wang X, Yang J, Yang S, Jiang Y. Biotechnol Bioeng. 2018 Sep 10. doi: 10.1002/bit.26827. 10.1002/bit.26827 PubMed 30199094 |
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 pYES2-gRNA-hyg-MCS 6007 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION E. coli-S. cerevisiae shuttle plasmid harbors gRNA expression
cassettes targeting S. cerevisiae chromosomal X-3 site (HXK1 gene).
Can function as a gRNA expression empty backbone plasmid..
ACCESSION .
VERSION .
KEYWORDS pYES2-gRNA-hyg-MCS
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6007)
AUTHORS Wang X, Yang J, Yang S, Jiang Y
TITLE Unraveling the genetic basis of fast l-arabinose consumption on top
of recombinant xylose-fermenting Saccharomyces cerevisiae.
JOURNAL Biotechnol Bioeng. 2018 Sep 10. doi: 10.1002/bit.26827.
PUBMED 30199094
REFERENCE 2 (bases 1 to 6007)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6007
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter complement(682..1025)
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
promoter 1032..1300
/label=SNR52 promoter
/note="promoter for the S. cerevisiae small nucleolar RNA
gene SNR52"
misc_RNA 1321..1395
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
terminator 1400..1419
/label=SUP4 terminator
/note="transcription terminator for the S. cerevisiae SUP4
tRNA gene"
primer_bind complement(1586..1603)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1757..2345)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1837..1856)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2516..3376)
/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 3139..3158
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4077..4954
/label=2u ori
/note="yeast 2u plasmid origin of replication"
primer_bind complement(5160..5181)
/label=F1ori-F
/note="F1 origin, forward primer"
gene complement(5457..1025)
/label=hphMX6
/note="yeast selectable marker conferring hygromycin
resistance"
terminator 5457..5654
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
CDS complement(5660..681)
/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"
ORIGIN
1 ttgggaatcc ccgaacatcg cctcgctcca gtcaatgacc gctgttatgc ggccattgtc
61 cgtcaggaca ttgttggagc cgaaatccgc gtgcacgagg tgccggactt cggggcagtc
121 ctcggcccaa agcatcagct catcgagagc ctgcgcgacg gacgcactga cggtgtcgtc
181 catcacagtt tgccagtgat acacatgggg atcagcaatc gcgcatatga aatcacgcca
241 tgtagtgtat tgaccgattc cttgcggtcc gaatgggccg aacccgctcg tctggctaag
301 atcggccgca gcgatcgcat ccatggcctc cgcgaccggc tgcagaacag cgggcagttc
361 ggtttcaggc aggtcttgca acgtgacacc ctgtgcacgg cgggagatgc aataggtcag
421 gctctcgctg aattccccaa tgtcaagcac ttccggaatc gggagcgcgg ccgatgcaaa
481 gtgccgataa acataacgat ctttgtagaa accatcggcg cagctattta cccgcaggac
541 atatccacgc cctcctacat cgaagctgaa agcacgagat tcttcgccct ccgagagctg
601 catcaggtcg gagacgctgt cgaacttttc gatcagaaac ttctcgacag acgtcgcggt
661 gagttcaggc tttttaccca tggttgttta tgttcggatg tgatgtgaga actgtatcct
721 agcaagattt taaaaggaag tatatgaaag aagaacctca gtggcaaatc ctaacctttt
781 atatttctct acaggggcgc ggcgtgggga caattcaacg cgtctgtgag gggagcgttt
841 ccctgctcgc aggtccgcag cgaggagccg taatttttgc ttcgcgccgt gcggccatca
901 aaatgtatgg atgcaaatga ttatacatgg ggatgtatgg gctaaatgta cgggcgacag
961 tcacatcatg cccctgagct gcgcacgtca agactgtcaa ggagggtatt ctgggcctcc
1021 atgtcactag ttctttgaaa agataatgta tgattatgct ttcactcata tttatacaga
1081 aacttgatgt tttctttcga gtatatacaa ggtgattaca tgtacgtttg aagtacaact
1141 ctagattttg tagtgccctc ttgggctagc ggtaaaggtg cgcatttttt cacaccctac
1201 aatgttctgt tcaaaagatt ttggtcaaac gctgtagaag tgaaagttgg tgcgcatgtt
1261 tcggcgttcg aaacttctcc gcagtgaaag ataaatgatc aatttcatcc atcaattcct
1321 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
1381 ggcaccgagt cggtggtgct ttttttgttt tttatgtcta gatcctagga tccaagcttg
1441 gtaccgagct cgtcgacgaa ttctgcagat atccatcaca ctggcggccg ctcgagcatg
1501 catctagagg gccgcaggcc ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
1561 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
1621 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
1681 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
1741 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
1801 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
1861 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
1921 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
1981 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
2041 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
2101 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
2161 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
2221 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
2281 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
2341 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
2401 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
2461 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
2521 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
2581 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
2641 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
2701 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
2761 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
2821 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
2881 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
2941 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
3001 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
3061 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
3121 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
3181 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
3241 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
3301 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
3361 aatgttgaat actcatactc ttcctttttc aatgggtaat aactgatata attaaattga
3421 agctctaatt tgtgagttta gtatacatgc atttacttat aatacagttt tttgaaaagc
3481 tagcttatcg atgataagct gtcaaacatg agaattaatt ccacggacta tagactatac
3541 ctagtatact ccgtctactg tacgatacac ttccgctcag gtccttgtcc tttaacgagg
3601 ccttaccact cttttgttac tctattgatc cagctcagca aaggcagtgt gatctaagat
3661 tctatcttcg cgatgtagta aaactagcta gaccgagaaa gagactagaa atgcaaaagg
3721 cacttctaca atggctgcca tcattattat ccgatgtgac gctgcagctt ctcaatgata
3781 ttcgaatacg ctttgaggag atacagccta atatccgaca aactgtttta cagatttacg
3841 atcgtacttg ttacccatca ttgaattttg aacatccgaa cctgggagtt ttccctgaaa
3901 cagatagtat atttgaacct gtataataat atatagtcta gcgctttacg gaagacaatg
3961 tatgtatttc ggttcctgga gaaactattg catctattgc ataggtaatc ttgcacgtcg
4021 catccccggt tcattttctg cgtttccatc ttgcacttca atagcatatc tttgttaacg
4081 aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa
4141 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc
4201 aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt
4261 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt
4321 ttaccaacaa agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta
4381 tttttctaac aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt
4441 ctcttgataa ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc
4501 tattttctct tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga
4561 agctgcgggt gcattttttc aagataaagg catccccgat tatattctat accgatgtgg
4621 attgcgcata ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa
4681 ttatgaacgg tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt
4741 cgtattgttt tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt
4801 aatactagag ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg
4861 aaaggtggat gggtaggtta tatagggata tagcacagag atatatagca aagagatact
4921 tttgagcaat gtttgtggaa gcggtattcg caatgggaag ctccaccccg gttgataatc
4981 agaaaagccc caaaaacagg aagattgtat aagcaaatat ttaaattgta agcgttaata
5041 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
5101 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
5161 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
5221 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
5281 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
5341 ggggaaagcc ctgatgaata tcttgtggta ggggtttggg aaaatcattc gagtttgatg
5401 tttttcttgg tatttcccac tcctcttcag agtacagaag attaagtgag agatcccagt
5461 atagcgacca gcattcacat acgattgacg catgatatta ctttctgcgc acttaacttc
5521 gcatctgggc agatgatgtc gaggcgaaaa aaaatataaa tcacgctaac atttgattaa
5581 aatagaacaa ctacaatata aaaaaactat acaaatgaca agttcttgaa aacaagaatc
5641 tttttattgt cagtactgat tattcctttg ccctcggacg agtgctgggg cgtcggtttc
5701 cactatcggc gagtacttct acacagccat cggtccagac ggccgcgctt ctgcgggcga
5761 tttgtgtacg cccgacagtc ccggctccgg atcggacgat tgcgtcgcat cgaccctgcg
5821 cccaagctgc atcatcgaaa ttgccgtcaa ccaagctctg atagagttgg tcaagaccaa
5881 tgcggagcat atacgcccgg agccgcggcg atcctgcaag ctccggatgc ctccgctcga
5941 agtagcgcgt ctgctgctcc atacaagcca accacggcct ccagaagaag atgttggcga
6001 cctcgta
//
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