| 質粒名稱: | pADH143 |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 90989 |
| 存儲實驗室: | Vladislav Verkhusha |
|
載體骨架基本信息 |
|
| 載體名稱: | pN1 |
| 載體出品公司: | Clontech |
| 空載體大小: | 4038 bp |
| 完整質粒大小: | 4989 bp |
| 載體修飾: | 人細胞色素c氧化酶亞基VIII（human cytochrome C oxidase subunit VIII） |
| 載體類型: | 哺乳細胞表達 |
| 篩選標記: | 新霉素（G418） |
| 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: | iRFP713 |
| 物種來源: | 光合細菌 Rhodopseudomonas palustris |
| 插入大小: | 951 bp |
| 基因突變: | T2A, S13L, A92T, V104T, V114I, E161K, Y193K, F198Y, D202T, I203V, Y258F, A283V, K288T, N290Y relative to RpBphP2 |
| GenBank編號 | JN247409 |
| 啟動子: | CMV |
| 標簽/融合蛋白: | 人細胞色素c氧化酶亞基VIII（N端） |
| 克隆方法: | 限制性內切酶 |
| 5’克隆位點: | AgeI(未破壞) |
| 3’克隆位點: | NotI (未破壞) |
| 5’測序引物 | CMV-F |
| 3’測序引物 | SV40pA-R |
|
文獻引用方法 |
|
| 材料和方法部分: | |
| 參考文獻部分: | |
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pADH143載體質粒基本信息
pADH143質粒圖譜載體圖譜和pADH143載體序列質粒序列多克隆位點信息
pADH143質粒載體簡介
pADH143質粒序列載體序列
LOCUS pADH143 4150 bp DNA circular SYN 14-APR-2025
DEFINITION NAT-marked ""empty"" gRNA expression construct; part 2 of 2 of C.mal
LEUpOUT CRISPR system. Use with pADH140 CAS9 expression construct..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4150)
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 4150)
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: pADH143
FEATURES Location/Qualifiers
source 1..4150
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 111..130
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(349..453)
/gene="bla"
/label=AmpR promoter
primer_bind 521..539
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
terminator 1110..1307
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
misc_RNA 1629..1704
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter complement(join(2525..2531,2532..2549,2550..2555))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2570..2591
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2708..2725)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2879..3467)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2959..2978)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(3638..4150,1..279,280..348))
/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"
ORIGIN
1 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
61 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
121 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
181 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
241 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
301 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
361 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
421 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
481 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
541 ctttgacggc acggccacgc gtttaaaccg ccaggtgatg ctgaagctat tgaagctttg
601 gatggttctt tcactaccga tactgttttt agagttactg ctactggtga tggtttcact
661 ttgagagaag ttccagttga tccaccattg actaaggttt tcccagatga tgaatccgat
721 gatgaatccg atgctggtga agatggtgat ccagattcta gaactttcgt tgcttatggt
781 gatgatggtg atttggctgg tttcgttgtt gtttcttatt ctggttggaa cagaagattg
841 actgttgaag atattgaagt tgctccagaa catagaggtc atggtgttgg tagagctttg
901 atgggtttgg ctactgaatt cgccagagaa agaggtgctg gtcatttgtg gttggaagtt
961 accaatgtta atgctccagc tattcatgct tatagaagaa tgggtttcac tttgtgtggt
1021 ttggatactg ctttatacga tggtactgct tccgatggtg aacaagcttt gtatatgtct
1081 atgccatgtc cataataact cgagtcagta ctgacaataa aaagattctt gttttcaaga
1141 acttgtcatt tgtatagttt ttttatattg tagttgttct attttaatca aatgttagcg
1201 tgatttatat tttttttcgc ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga
1261 aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat
1321 actaacatct aatcaactcc cagatcacca ctatacattt aaaagtcatt ggtcaataac
1381 tatactcgag tattgcctca tcaaagaaac aatcaaatat tatagatact cactccatca
1441 cgtgataatt tcactggtat ggaaaagtgg aaaattttat aaaaaaaaat ttgatgcctt
1501 tggcatagct gaaacttcgg cccaatagga ttggagaata tgttttcgca gcgttcttac
1561 aattaaattg tggtggaagt tcgagacttg cgtaaactat ttttaatttg gagacggaat
1621 tccgtctcgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
1681 gaaaaagtgg caccgagtcg gtgctttttt tctcgagttt ttttatcgag ttgagaccag
1741 aagtcgttaa aggtacaaac ttgatcattg ttcgtgaatt agttggtggt atttatttcg
1801 gtgatcgtga agaacaagaa gaaagtgaag ataaacaaac cgcatgggat actgaaaaat
1861 acaccgtcga tgaagtcacc agaatcaccc gtatggctgc cttcatggct ttacaacata
1921 acccaccatt accaatctgg tcattggata aggctaatgt tttagcttct tctagattat
1981 ggagaagaac tgtcgataaa gtgatttctg aagagttccc aactttgtct gttcaacatc
2041 aattgattga ctccgccgcc atgattttaa ttcaaaaccc aaccaaattg aatggtataa
2101 tcatcacttc taacatgttt ggtgatatca tttccgatga agcttcagtt attccaggtt
2161 ccttgggttt attaccatct gcatctttgg cttcattgcc agataccaac accgcttttg
2221 gtctttatga accatgtcac ggttctgctc ctgatttacc agccaataaa gttaacccaa
2281 ttgctactat tttatctgct gcttctatgt tgagattatc tttggattgt gtgaaagaag
2341 ctgaagcttt agaagaagcc gtgaaacaag tcttggataa aggtatcaga actgcagatt
2401 taagaggtac cagttcagct actgaagttg gtgacgcgat tgtcgaagct gttactaaaa
2461 tcttaaaaga aaaagcttaa cggtgtttaa accccagcgc ctggcgggcg ctcacaattc
2521 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
2581 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
2641 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
2701 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
2761 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
2821 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
2881 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
2941 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
3001 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
3061 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
3121 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
3181 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
3241 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
3301 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
3361 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
3421 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
3481 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
3541 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
3601 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgcttg atcagtgagg
3661 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
3721 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
3781 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
3841 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
3901 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
3961 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
4021 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
4081 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
4141 attctcttac
//
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