| 质粒名称: | pADH139 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 90987 |
| 存储实验室: | Aaron Hernday |
|
载体骨架基本信息 |
|
| 载体名称: | pUC19 |
| 载体出品公司: | Clontech |
| 空载体大小: | 2242bp |
| 完整质粒大小: | 3094bp |
| 载体类型: | CRISPR |
| 筛选标记: | Nourseothricin (NAT) |
| 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: | gRNA conserved, C, mal LEU2 2 of 2 |
|
文献引用方法 |
|
| 材料和方法部分: | pADH139 was a gift from Aaron Hernday (Addgene plasmid # 90987 ; http://n2t.net/addgene:90987 ; RRID:Addgene_90987) |
| 参考文献部分: | 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 |
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pADH139载体质粒基本信息
pADH139质粒图谱载体图谱和pADH139载体序列质粒序列多克隆位点信息
pADH139质粒载体简介
pADH139质粒序列载体序列
LOCUS pADH139 3094 bp DNA circular SYN 14-APR-2025
DEFINITION NAT-marked ""empty"" gRNA construct for use in gRNA expression
cassette stitching PCR; use with pADH110 to generate target-specific
part 2 of 2 for C.mal LEUpOUT CRISPR system..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3094)
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 3094)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
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: pADH139
FEATURES Location/Qualifiers
source 1..3094
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 138..156
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
misc_RNA 190..265
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter complement(join(1086..1092,1093..1110,1111..1116))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1131..1152
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1269..1286)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1440..2028)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1520..1539)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(2199..2990,2991..3059))
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATIDERNRQIAEIGAS
LIKHW"
primer_bind 2822..2841
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(join(3060..3094,1..70))
/gene="bla"
/label=AmpR promoter
ORIGIN
1 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
61 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
121 tgacattaac ctataaaaat aggcgtatca cgaggccctt tgacggcacg gccacgcgtt
181 taaaccgccg ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
241 tgaaaaagtg gcaccgagtc ggtgcttttt ttctcgagtt tttttatcga gttgagacca
301 gaagtcgtta aaggtacaaa cttgatcatt gttcgtgaat tagttggtgg tatttatttc
361 ggtgatcgtg aagaacaaga agaaagtgaa gataaacaaa ccgcatggga tactgaaaaa
421 tacaccgtcg atgaagtcac cagaatcacc cgtatggctg ccttcatggc tttacaacat
481 aacccaccat taccaatctg gtcattggat aaggctaatg ttttagcttc ttctagatta
541 tggagaagaa ctgtcgataa agtgatttct gaagagttcc caactttgtc tgttcaacat
601 caattgattg actccgccgc catgatttta attcaaaacc caaccaaatt gaatggtata
661 atcatcactt ctaacatgtt tggtgatatc atttccgatg aagcttcagt tattccaggt
721 tccttgggtt tattaccatc tgcatctttg gcttcattgc cagataccaa caccgctttt
781 ggtctttatg aaccatgtca cggttctgct cctgatttac cagccaataa agttaaccca
841 attgctacta ttttatctgc tgcttctatg ttgagattat ctttggattg tgtgaaagaa
901 gctgaagctt tagaagaagc cgtgaaacaa gtcttggata aaggtatcag aactgcagat
961 ttaagaggta ccagttcagc tactgaagtt ggtgacgcga ttgtcgaagc tgttactaaa
1021 atcttaaaag aaaaagctta acggtgttta aaccccagcg cctggcgggc gctcacaatt
1081 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
1141 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
1201 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
1261 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
1321 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
1381 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
1441 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
1501 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
1561 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
1621 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
1681 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
1741 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
1801 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
1861 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
1921 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
1981 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
2041 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
2101 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
2161 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt gatcagtgag
2221 gcacctatct cagcgatctg tctatttcgt tcatcaatag ttgcctgact ccccgtcgtg
2281 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
2341 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
2401 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
2461 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
2521 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
2581 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
2641 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
2701 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
2761 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
2821 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
2881 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
2941 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
3001 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
3061 ctcttccttt ttcaatatta ttgaagcatt tatc
//
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