| 质粒名称: | pCE27 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 174405 |
| 存储实验室: | Clarissa Nobile |
|
载体骨架基本信息 |
|
| 载体名称: | pUC19 |
| 载体出品公司: | -- |
| 空载体大小: | 2242 bp |
| 完整质粒大小: | 4307 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达,CRISPR |
| 筛选标记: | Nourseothricin (NAT) |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 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: | NAT 2 of 2, pSNR52, ADE2 gRNA, gRNA conserved, C. auris LEU2 2 of 2 |
| 物种来源: | Candida auris |
| 插入大小: | 2070 bp |
| 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: | pCE27 was a gift from Clarissa Nobile (Addgene plasmid # 174405 ; http://n2t.net/addgene:174405 ; RRID:Addgene_174405) |
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 pCE27 4304 bp ds-DNA circular SYN 08-DEC-2021
DEFINITION C. auris LEUpOUT NAT marker gRNA expression construct. Use with
pCE35 CAS9 expression construct..
ACCESSION .
VERSION .
KEYWORDS pCE27
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4304)
TITLE Plasmids for markerless genome editing in C. auris
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 4304)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Dec 8, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4304
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(27..887)
/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 650..669
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(888..992)
/gene="bla"
/label=AmpR promoter
primer_bind 1060..1078
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
terminator 1649..1846
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
misc_RNA 2322..2397
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter complement(3218..3248)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3263..3284
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3401..3418)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3572..4160)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3652..3671)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 atatatgagt aaacttggtc tgacagttac caatgcttga tcagtgaggc acctatctca
61 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
121 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
181 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
241 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
301 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
361 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
421 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
481 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
541 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
601 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
661 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
721 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
781 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
841 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
901 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
961 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
1021 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
1081 tttgacggca cggccacgcg tttaaaccgc caggtgatgc tgaagctatt gaagctttgg
1141 atggttcttt cactaccgat actgttttta gagttactgc tactggtgat ggtttcactt
1201 tgagagaagt tccagttgat ccaccattga ctaaggtttt cccagatgat gaatccgatg
1261 atgaatccga tgctggtgaa gatggtgatc cagattctag aactttcgtt gcttatggtg
1321 atgatggtga tttggctggt ttcgttgttg tttcttattc tggttggaac agaagattga
1381 ctgttgaaga tattgaagtt gctccagaac atagaggtca tggtgttggt agagctttga
1441 tgggtttggc tactgaattc gccagagaaa gaggtgctgg tcatttgtgg ttggaagtta
1501 ccaatgttaa tgctccagct attcatgctt atagaagaat gggtttcact ttgtgtggtt
1561 tggatactgc tttatacgat ggtactgctt ccgatggtga acaagctttg tatatgtcta
1621 tgccatgtcc ataataactc gagtcagtac tgacaataaa aagattcttg ttttcaagaa
1681 cttgtcattt gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt
1741 gatttatatt ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa
1801 agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt cgattcgata
1861 ctaaagctta acttttagag tcacttaccc tacaaaggtg taagcttccc tcagatcagg
1921 cggcacgcat ctgatatcta acatccattg taaaattagt gaagattaac actcagagcc
1981 agttattttg aacacagatg gcttttggct gctcttatcg tctacgtttc tttttttttt
2041 tttttttttt ttttttccta attttttacg agttcactaa ttggtttcgg gggctcaatt
2101 ttcgaatgcg acattcataa actttgacct tttatgtgtt gatctggata attttgacct
2161 caaaactcgg aaaaattgat gtatagcgta ttcgacatgg ctttaaaagt cgcagagttc
2221 tacgattttc atttgcactg gcccatccgg taagcctcgg ggctccggca tgtgagggtt
2281 cgactccctc agcagaaaac atgccgtgaa agaaaacacc agttttagag ctagaaatag
2341 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt
2401 ttttctcgag tttttttatc gagtctccat tgaaggccag tgttgtcaaa ggaaccaatt
2461 tcaccgtggt gcgtgaattg gtcggtggta tctactttgg tgagagacaa gaacaggaag
2521 agtctgccga cggtcagagc gcttgggaca ccgagaagta cagtgtctct gaagttgaaa
2581 gaatcaccag aatggctgcc ttcatggctt tgcagcacaa ccctcctttg cccatctggt
2641 ccttagacaa ggccaacgtg cttgcctcat ctagattgtg gagaaagacg gtcgacaagg
2701 tgatgagcga ggagttccct caattgacca tcaaccacca gctcattgac tctgctgcca
2761 tgattttggt gcagagccca actaaattga acggtgttgt gatcacgtca aacatgtttg
2821 gtgatattat ttccgacgag gcttccgtga tccctggctc cttgggtctt ttaccttctg
2881 catctttggc ttcattacca gatacaaata aggcgtttgg tttgtacgaa ccatgccacg
2941 gttcggctcc ggacttgcca gaaaacaagg tcaaccctgt cgccacgatt ttgtcggccg
3001 ctatgatgtt gcgcttgtca ttagactgcc tcagagaggc agaggccttg gagaaggctg
3061 ttcgtaacgt tttggactct ggtattcgaa ccgcagattt gaagggctca agttccactc
3121 aggaagtggg cgatgccatt gctgctgagg taaagaagct tttgcagaaa taacggtgtt
3181 taaaccccag cgcctggcgg gcgctcacaa ttccacacaa catacgagcc ggaagcataa
3241 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
3301 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
3361 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
3421 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
3481 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
3541 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
3601 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
3661 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
3721 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
3781 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
3841 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
3901 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
3961 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
4021 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
4081 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
4141 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
4201 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
4261 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagt
//
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