| 質粒名稱: | pADH100 |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 90980 |
| 存儲實驗室: | Aaron Hernday |
|
載體骨架基本信息 |
|
| 載體名稱: | ppUC191 |
| 載體出品公司: | Clontech |
| 空載體大小: | 2242 bp |
| 完整質粒大小: | 3934 bp |
| 載體類型: | CRISPR |
| 篩選標記: | Nourseothricin (NAT) |
| 載體抗性: | 氨芐霉素 |
| 生長溫度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷貝數: | 高拷貝 |
|
插入基因信息 |
|
| 插入基因名稱: |
NAT 2 of 2, pSNR52, empty gRNA, gRNA conserved, FRT, DS_HIS1
gRNA/shRNA sequencenone
|
|
文獻引用方法 |
|
| 材料和方法部分: | pADH100 was a gift from Aaron Hernday (Addgene plasmid # 90980 ; //n2t.net/addgene:90980 ; RRID:Addgene_90980) |
| 參考文獻部分: | An Efficient, Rapid, and Recyclable System for CRISPR-Mediated Genome Editing in Candida albicans. Nguyen N, Quail MMF, Hernday AD. mSphere. 2017 Apr 26;2(2):e00149-17. doi: 10.1128/mSphereDirect.00149-17. eCollection 2017 Mar-Apr. 10.1128/mSphereDirect.00149-17 PubMed 28497115 |
買家導航
pADH100載體質粒基本信息
pADH100質粒圖譜載體圖譜和pADH100載體序列質粒序列多克隆位點信息
pADH100質粒載體簡介
pADH100質粒序列載體序列
LOCUS pADH100 3934 bp DNA circular SYN 14-APR-2025
DEFINITION NAT-marked ""empty"" gRNA expression construct; part 2 of 2 of C.alb
HIS1-FLP CRISPR system. Use with pADH99 CAS9 expression construct..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3934)
AUTHORS Nguyen N, Quail MMF, Hernday AD
TITLE An Efficient, Rapid, and Recyclable System for CRISPR-Mediated
Genome Editing in Candida albicans.
JOURNAL mSphere. 2017 Apr 26;2(2):e00149-17. doi:
10.1128/mSphereDirect.00149-17. eCollection 2017 Mar-Apr.
PUBMED 28497115
REFERENCE 2 (bases 1 to 3934)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
//gt12365.cn
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1128/mSphereDirect.00149-17"; journalName: "mSphere"; date:
"2017-04-26- 26"; volume: "2"; issue: "2"; pages: "e00149-17"
COMMENT Sequence Label: pADH100
FEATURES Location/Qualifiers
source 1..3934
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 4..22
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
terminator 593..790
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
misc_RNA 1112..1187
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
protein_bind complement(1214..1247)
/label=FRT (minimal)
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
promoter complement(join(1792..1798,1799..1816,1817..1822))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1837..1858
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1975..1992)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2146..2734)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2226..2245)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(2905..3696,3697..3765))
/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 3528..3547
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3766..3870)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 aaaaataggc gtatcacgag gccctttgac ggcacggcca cgcgtttaaa ccgccaggtg
61 atgctgaagc tattgaagct ttggatggtt ctttcactac cgatactgtt tttagagtta
121 ctgctactgg tgatggtttc actttgagag aagttccagt tgatccacca ttgactaagg
181 ttttcccaga tgatgaatcc gatgatgaat ccgatgctgg tgaagatggt gatccagatt
241 ctagaacttt cgttgcttat ggtgatgatg gtgatttggc tggtttcgtt gttgtttctt
301 attctggttg gaacagaaga ttgactgttg aagatattga agttgctcca gaacatagag
361 gtcatggtgt tggtagagct ttgatgggtt tggctactga attcgccaga gaaagaggtg
421 ctggtcattt gtggttggaa gttaccaatg ttaatgctcc agctattcat gcttatagaa
481 gaatgggttt cactttgtgt ggtttggata ctgctttata cgatggtact gcttccgatg
541 gtgaacaagc tttgtatatg tctatgccat gtccataata actcgagtca gtactgacaa
601 taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt
661 tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc
721 agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt
781 cgctatactg ctgtcgattc gatactaaca tctaatcaac tcccagatca ccactataca
841 tttaaaagtc attggtcaat aactatactc gagtattgcc tcatcaaaga aacaatcaaa
901 tattatagat actcactcca tcacgtgata atttcactgg tatggaaaag tggaaaattt
961 tataaaaaaa aatttgatgc ctttggcata gctgaaactt cggcccaata ggattggaga
1021 atatgttttc gcagcgttct tacaattaaa ttgtggtgga agttcgagac ttgcgtaaac
1081 tatttttaat ttggagacgg aattccgtct cgttttagag ctagaaatag caagttaaaa
1141 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttctcgag
1201 tttttttatc gaggaagttc ctatactttc tagagaatag gaacttccca ccttgatgta
1261 cacaccttta aagtcaatag gcattctcgt ttgtaattcc ttgacacaca tatccattaa
1321 ctttaaacca tcttcgaccg tcatgtcctt tctataatgt ttatctaaca aagatgttgt
1381 gtaaaatgca gcgtagccat gagcaccata agggagttca acttgtgtac ccaaataatc
1441 tatccaattc aacgatggct catttttctt tacatcatat cctccaatta aacagttcac
1501 ctggtacggt tttctagatc taagggaact ggccaattgg tttctcacaa acgaggctgt
1561 tgcctttgga gataactcaa tgtcattttc tctcatcgag tacaattgaa tgtttgcttg
1621 aacatactcg gcaaactgaa ccgtatcacc agcttcacca gtgtatgcta ttaagttgtg
1681 tgcatttaaa tgtcttgttt tatcatctgt atcctttaaa attgaaatac ctcttgtggc
1741 agctttacgg tgtttaaacc ccagcgcctg gcgggcgctc acaattccac acaacatacg
1801 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
1861 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
1921 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
1981 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
2041 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
2101 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
2161 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
2221 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
2281 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
2341 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
2401 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
2461 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
2521 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
2581 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
2641 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
2701 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
2761 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
2821 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
2881 atatgagtaa acttggtctg acagttacca atgcttgatc agtgaggcac ctatctcagc
2941 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
3001 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
3061 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
3121 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
3181 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
3241 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
3301 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
3361 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
3421 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
3481 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
3541 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
3601 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
3661 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
3721 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
3781 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
3841 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
3901 ctaagaaacc attattatca tgacattaac ctat
//