| 质粒名称: | DR274 ，pDR274 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 42250 |
| 存储实验室: | Keith Joung |
|
载体骨架基本信息 |
|
| 载体名称: | Zebrafish-gRNA-ExpressionVector |
| 载体出品公司: | Joung lab DR274 |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 2147 bp |
| 载体修饰: | -- |
| 载体类型: | 蠕虫表达式 |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 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: | DR274 was a gift from Keith Joung (Addgene plasmid # 42250 ; http://n2t.net/addgene:42250 ; RRID:Addgene_42250) |
| 参考文献部分: | Efficient genome editing in zebrafish using a CRISPR-Cas system. Hwang WY, Fu Y, Reyon D, Maeder ML, Tsai SQ, Sander JD, Peterson RT, Yeh JR, Joung JK. Nat Biotechnol. 2013 Jan 29. doi: 10.1038/nbt.2501. 10.1038/nbt.2501 PubMed 23360964 |
买家导航
pDR274载体基本信息
pDR274载体图谱质粒图谱和多克隆位点信息
pDR274载体简介
pDR274载体序列
LOCUS DR274 2147 bp ds-DNA circular SYN 23-AUG-2019
DEFINITION guide RNA (gRNA) expression vector used to create a gRNA to a
specific sequence; uses T7 promoter for in vitro transcription.
ACCESSION .
VERSION .
KEYWORDS DR274
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2147)
AUTHORS Hwang WY, Fu Y, Reyon D, Maeder ML, Tsai SQ, Sander JD, Peterson RT,
Yeh JR, Joung JK
TITLE Efficient genome editing in zebrafish using a CRISPR-Cas system.
JOURNAL Nat Biotechnol. 2013 Jan 29. doi: 10.1038/nbt.2501.
PUBMED 23360964
REFERENCE 2 (bases 1 to 2147)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 23, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..2147
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 9..26
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 10..26
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_RNA complement(208..283)
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter complement(305..323)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(452..468)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(452..468)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
terminator 516..547
/label=tonB terminator
/note="bidirectional E. coli tonB-P14 transcription
terminator"
promoter 548..650
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS 651..1466
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCLKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(724..743)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
terminator 1490..1517
/label=T7Te terminator
/note="phage T7 early transcription terminator"
rep_origin complement(1529..2116)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1609..1628)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cccacccgtg taaaacgacg gccagtttat ctagtcagct tgattctagc tgatcgtgga
61 ccggaaggtg agccagtgag ttgattgcag tccagttacg ctggagtctg aggctcgtcc
121 tgaatgatat gcgaccgccg gagggttgcg tttgagacgg gcgacagatc cagtcgcgct
181 gctctcgtcg atccaagctt tttaaaagca ccgactcggt gccacttttt caagttgata
241 acggactagc cttattttaa cttgctattt ctagctctaa aactgagacc ctctctcggt
301 ctctcctata gtgagtcgta ttagctagcg gtgcgagcgg atcgagcagt gtcgatcact
361 actggaccgc gagctgtgct gcgacccgtg atcttacggc attatacgta tgatcggtcc
421 acgatcagct agattatcta gtcagcttga tgtcatagct gtttcctgag gctcaatact
481 gaccatttaa atcatacctg acctccatag cagaaagtca aaagcctccg accggaggct
541 tttgacttga tcggcacgta agaggttcca actttcacca taatgaaata agatcactac
601 cgggcgtatt ttttgagtta tcgagatttt caggagctaa ggaagctaaa atgagccata
661 ttcaacggga aacgtcttgc ttgaagccgc gattaaattc caacatggat gctgatttat
721 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt
781 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
841 atgttacaga tgagatggtc aggctaaact ggctgacgga atttatgcct cttccgacca
901 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atcccaggga
961 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
1021 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacggcg
1081 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttggtgcga
1141 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
1201 aactcttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
1261 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
1321 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
1381 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
1441 acttgatgct cgatgagttt ttctaatgag gacctaaatg taatcacctg gctcaccttc
1501 gggtgggcct ttctgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
1561 acaaaaatcg atgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
1621 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
1681 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
1741 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
1801 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
1861 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
1921 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
1981 gtatctgcgc tctgctgaag ccagttacct cggaaaaaga gttggtagct cttgatccgg
2041 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
2101 aaaaaaagga tctcaagaag atcctttgat tttctaccga agaaagg
//
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