| 质粒名称: | pcU6_3 sgRNA |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 92359 |
| 存储实验室: | Tatjana Sauka-Spengler |
|
载体骨架基本信息 |
|
| 载体名称: | pNG3 |
| 载体出品公司: | -- |
| 载体抗性: | |
| 空载体大小: | 3017 bp |
| 完整质粒大小: | 3494 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 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: | chick U6.3 promoter and gRNA cloning cassette |
| 物种来源: | -- |
| 插入大小: | 477 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pcU6_3 sgRNA was a gift from Tatjana Sauka-Spengler (Addgene plasmid # 92359 ; http://n2t.net/addgene:92359 ; RRID:Addgene_92359) |
| 参考文献部分: | Genome and epigenome engineering CRISPR toolkit for in vivo modulation of cis-regulatory interactions and gene expression in the chicken embryo. Williams RM, Senanayake U, Artibani M, Taylor G, Wells D, Ahmed AA, Sauka-Spengler T. Development. 2018 Feb 23;145(4). pii: dev.160333. doi: 10.1242/dev.160333. 10.1242/dev.160333 PubMed 29386245 |
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 pcU6_3 sgRNA 3494 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Chicken-specific U6 sgRNA expression mini-vector, harbouring chick
U6_3 pol III promoter..
ACCESSION .
VERSION .
KEYWORDS pcU6_3 sgRNA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3494)
AUTHORS Williams RM, Senanayake U, Artibani M, Taylor G, Wells D, Ahmed AA,
Sauka-Spengler T
TITLE Genome and epigenome engineering CRISPR toolkit for probing in vivo
cis-regulatory interactions in the chicken embryo
JOURNAL BioRxiv
REFERENCE 2 (bases 1 to 3494)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3494
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 196..224
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
CDS 272..1462
/codon_start=1
/gene="tet"
/product="tetracycline efflux protein"
/label=TcR
/note="confers resistance to tetracycline"
/translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
primer_bind complement(624..643)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind complement(1636..1655)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1637..1655)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1660..1676)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1660..1676)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
rep_origin 2343..2931
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2832..2851
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
terminator 3261..3288
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 3380..3466
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
ORIGIN
1 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
61 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
121 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
181 aattaattct catgtttgac agcttatcat cgattagctt taatgcggta gtttatcaca
241 gttaaattgc taacgcagtc aggcaccgtg tatgaaatct aacaatgcgc tcatcgtcat
301 cctcggcacc gtcaccctgg atgctgtagg cataggcttg gttatgccgg tactgccggg
361 cctcttgcgg gatatcgtcc attccgacag catcgccagt cactatggcg tgctgctagc
421 gctatatgcg ttgatgcaat ttctatgcgc acccgttctc ggagcactgt ccgaccgctt
481 tggccgccgc ccagtcctgc tcgcttcgct acttggagcc actatcgact acgcgatcat
541 ggcgaccaca cccgtcctgt ggattctcta cgccggacgc atcgtggccg gcatcaccgg
601 cgccacaggt gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc
661 tcgccacttc gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc
721 cgggggactg ttgggcgcca tctccttaca tgcaccattc cttgcggcgg cggtgctcaa
781 cggcctcaac ctactactgg gctgcttcct aatgcaggag tcgcataagg gagagcgccg
841 acccatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
901 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc
961 agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct
1021 gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc
1081 caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccg acgcgctggg
1141 ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc ttccccatta tgattcttct
1201 cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga
1261 cgaccatcag ggacagcttc aaggatcgct cgcggctctt accagcctaa cttcgatcat
1321 tggaccgctg atcgtcacgg cgatttatgc cgcctcggcg agcacatgga acgggttggc
1381 atggattgta ggcgccgccc tataccttgt ctgcctcccc gcgttgcgtc gcggtgcatg
1441 gagccgggcc acctcgacct gaatggaagc cggcggcacc tcgctaacgg attcaccact
1501 ccaagaattg gagccaatca attcttgcgg agaactgtga atgcgcaaac caacccttga
1561 tcggggaaga acagtatgtc gagctatttt ttgacttact ggggatcaag cctgattggg
1621 agaaaataaa atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag
1681 ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca
1741 tgcctcctct agaggtctca ggaccgcggc cgctccttca ggcactgccg acgacagccc
1801 aggcggaggt cctgagcgcc ggcgccaaat ttgcataaag agctacccag aagccctcgc
1861 gcgcggaaac gggcaaaaaa gggcttctaa tatggaaata ttacgccgaa tcgcgtcaca
1921 aattggctaa gcgggcctaa gagttaacaa gatgtgctat taagcggagc cttttggtgg
1981 gaagaaacgg aacagtcact gtgttctaaa agaacttgca ggatgagcct ttaaataccg
2041 cagtctcgac gctcttagtc ggagacgttg atttcgtctc tgtttaagag ctatgctgga
2101 aacagcatag caagtttaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
2161 tcggtgcttt tttgttttcc atattttttt cgcattgctt tccttggctt ttacaccctt
2221 acgcagcctc gccccctttt cccagggaga ccctcgagcc acccatgacc aaaatccctt
2281 aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
2341 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
2401 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
2461 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
2521 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
2581 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
2641 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
2701 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
2761 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
2821 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
2881 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
2941 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
3001 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
3061 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
3121 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
3181 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac cgctagccag
3241 gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttagtttgat
3301 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt
3361 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
3421 taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc ctggcagttc
3481 cctactctcg cgtt
//
