| 质粒名称: | pGTR |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 63143 |
| 存储实验室: | Yinong Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pGEM-T Easy |
| 载体出品公司: | Promega |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 3015 bp |
| 完整质粒大小: | 3192 bp |
| 载体修饰: | 通过TA克隆插入gRNA-tRNA融合片段 |
| 载体类型: | CRISPR，合成生物学 |
| 筛选标记: | -- |
| 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: | gRNA-tRNA fusion,GTR |
| 物种来源: | Synthetic |
| 插入大小: | 153 bp |
| 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: | pGTR was a gift from Yinong Yang (Addgene plasmid # 63143 ; http://n2t.net/addgene:63143 ; RRID:Addgene_63143) |
| 参考文献部分: | Boosting CRISPR/Cas9 multiplex editing capability with the endogenous tRNA-processing system. Xie K, Minkenberg B, Yang Y. Proc Natl Acad Sci U S A. 2015 Mar 17;112(11):3570-5. doi: 10.1073/pnas.1420294112. Epub 2015 Mar 2. 10.1073/pnas.1420294112 PubMed 25733849 |
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 pGTR 3192 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION PCR template to construct polycistronic tRNA-gRNA.
ACCESSION .
VERSION .
KEYWORDS pGTR
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3192)
AUTHORS Xie K, Minkenberg B, Yang Y
TITLE Boosting CRISPR/Cas9 multiplex editing capability with the
endogenous tRNA-processing system.
JOURNAL Proc Natl Acad Sci U S A. 2015 Mar 17;112(11):3570-5. doi:
10.1073/pnas.1420294112. Epub 2015 Mar 2.
PUBMED 25733849
REFERENCE 2 (bases 1 to 3192)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3192
/organism="synthetic DNA construct"
/mol_type="other DNA"
regulatory 33..42
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
regulatory 37..46
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
misc_RNA complement(145..220)
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter complement(317..335)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(318..335)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(353..369)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(353..369)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(366..388)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 377..393
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(401..431)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 446..467
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(584..601)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(755..1343)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(835..854)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1514..2374)
/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 2137..2156
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2375..2479)
/gene="bla"
/label=AmpR promoter
rep_origin complement(2557..3012)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2694..2715)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2906..2925
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3138..3160
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 3152..3169
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 3153..3169
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3176..3
/label=T7
/note="T7 promoter, forward primer"
promoter 3176..2
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 gggcgaattg ggcccgacgt cgcatgctcc cggccgccat ggcggccgcg ggaattcgat
61 tctgccatgc accagccggg aatcgaaccc gggtctgtac cgtggcaggg tactattcta
121 ccactagacc actggtgctt tgttgcaccg actcggtgcc actttttcaa gttgataacg
181 gactagcctt attttaactt gctatttcta gctctaaaac cgagacctcg gtctccaatc
241 actagtgaat tcgcggccgc ctgcaggtcg accatatggg agagctccca acgcgttgga
301 tgcatagctt gagtattcta tagtgtcacc taaatagctt ggcgtaatca tggtcatagc
361 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
421 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
481 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
541 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
601 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
661 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
721 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
781 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
841 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
901 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
961 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
1021 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
1081 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
1141 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
1201 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
1261 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
1321 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
1381 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
1441 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
1501 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
1561 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
1621 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
1681 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
1741 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
1801 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
1861 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
1921 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
1981 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
2041 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
2101 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
2161 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
2221 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
2281 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
2341 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
2401 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
2461 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgatgcg gtgtgaaata
2521 ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt aatattttgt
2581 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
2641 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
2701 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
2761 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
2821 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
2881 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
2941 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
3001 tacagggcgc gtccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
3061 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
3121 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata
3181 cgactcacta ta
//
