| 质粒名称: | pCBC-MT1T2 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 50593 |
| 存储实验室: | Qi-Jun Chen |
|
载体骨架基本信息 |
|
| 载体名称: | pGWC |
| 载体出品公司: | -- |
| 载体抗性: | 氯霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 植物表达，CRISPR；PCR模板 |
| 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: | T1, gRNA scaffold, OsU3t plus, TaU3p, T2 |
| 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 | SP6 |
| 3' 测序引物 | T7 |
|
文献引用方法 |
|
| 材料和方法部分: | pCBC-MT1T2 was a gift from Qi-Jun Chen (Addgene plasmid # 50593 ; http://n2t.net/addgene:50593 ; RRID:Addgene_50593) |
| 参考文献部分: | A CRISPR/Cas9 toolkit for multiplex genome editing in plants. Xing HL, Dong L, Wang ZP, Zhang HY, Han CY, Liu B, Wang XC, Chen QJ. BMC Plant Biol. 2014 Nov 29;14(1):327. 10.1186/s12870-014-0327-y PubMed 25432517 |
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 pCBC-MT1T2 3849 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION CRISPR/Cas based plant genome editing and gene regulation; used as
template for making expression cassette with multiple gRNA target
sites.
ACCESSION .
VERSION .
KEYWORDS pCBC-MT1T2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3849)
AUTHORS Xing HL, Dong L, Wang ZP, Zhang HY, Han CY, Liu B, Wang XC, Chen QJ
TITLE A CRISPR/Cas9 toolkit for multiplex genome editing in plants.
JOURNAL BMC Plant Biol. 2014 Nov 29;14(1):327.
PUBMED 25432517
REFERENCE 2 (bases 1 to 3849)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3849
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 158..817
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKSKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(224..243)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
rep_origin 1042..1630
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1531..1550
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
terminator 1795..1881
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 1973..2000
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
primer_bind 2008..2031
/label=pENTR-F
/note="pENTR vectors, forward primer"
promoter 2042..2089
/label=EM7 promoter
/note="synthetic bacterial promoter "
primer_bind 2109..2131
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 2128..2144
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2128..2144
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 2162..2180
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 2162..2179
/label=SP6
/note="SP6 promoter, forward primer"
misc_RNA 2235..2310
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
primer_bind complement(3176..3195)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3177..3195)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3202..3219)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(3202..3218)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3211..3233)
/label=M13/pUC Forward
/note="In lacZ gene"
CDS 3356..3658
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
primer_bind 3537..3556
/label=ccdB-fwd
/note="ccdB gene, forward primer"
ORIGIN
1 ggttttcatg gcttgttatg actgtttttt tggggtacag tctatgcctc gggcatccaa
61 gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac
121 gcagcagggc agtcgcccta aaacaaagtt aaacatcatg gagaaaaaaa tcactggata
181 taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt
241 tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt
301 aaagaaaagt aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa
361 tgctcatccg gagttccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt
421 tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga
481 ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg
541 tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa
601 tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc
661 ccccgttttc acaatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc
721 gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt
781 acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca gttattggtg
841 cgatcctcta gctagaaatt cgttcaagcc gacgccgctt cgcggcgcgg cttaactcaa
901 gcgttagatg cactaagcac ataattgctc acagccaaac tatcaggtca agtcccgcgg
961 agttgttcgg taaattgtca caacgccgcc cgccccgttc cactgagcgt cagacccggt
1021 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
1081 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
1141 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
1201 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
1261 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
1321 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
1381 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
1441 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
1501 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
1561 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
1621 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
1681 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgctag
1741 gatggatctc gggcacgtct aactactaag cgagagtagg gaactgccag gcatcaaata
1801 aaacgaaagg ctcagtcgga agactgggcc tttcgtttta tctgttgttt gtcggtgaac
1861 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgtgaa gcaacggccc
1921 ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaactaa gcagaaggcc
1981 atcctgacgg atggcctttt tgcgtttcta caaactcttc ctgttagtta gttacttaag
2041 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
2101 ctaaaccgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
2161 tatttaggtg acactataga atactcaagc tatgcaaggc ccttaaggcc aagggnnnnn
2221 ntgccaacca acgagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat
2281 caacttgaaa aagtggcacc gagtcggtgc tttttttttt cgttttgcat tgagttttct
2341 ccgtcgcatg tttgcagttt tattttccgt tttgcattga aatttctccg tctcatgttt
2401 gcagcgtgtt caaaaagtac gcagctgtat ttcacttatt tacggcgcca cattttcatg
2461 ccgtttgtgc caactatccc gagctagtga atacagcttg gcttcacaca acactggtga
2521 cccgctgacc tgctcgtacc tcgtaccgtc gtacggcaca gcatttggaa ttaaagggtg
2581 tgatcgatac tgcttgctgc tcatgaatcc aaaccacacg gagttcaaat tcccacagat
2641 taaggctcgt ccgtcgcaca aggtaatgtg tgaatattat atctgtcgtg caaaattgcc
2701 tggcctgcac aattgctgtt atagttggcg gcagggagag ttttaacatt gactagcgtg
2761 ctgataattt gtgagaaata ataattgaca agtagatact gacatttgag aagagcttct
2821 gaactgttat tagtaacaaa aatggaaagc tgatgcacgg aaaaaggaaa gaaaaagcca
2881 tacttttttt taggtaggaa aagaaaaagc catacgagac tgatgtctct cagatgggcc
2941 gggatctgtc tatctagcag gcagcagccc accaacctca cgggccagca attacgagtc
3001 cttctaaaag ctcccgccga ggggcgctgg cgctgctgtg cagcagcacg tctaacatta
3061 gtcccacctc gccagtttac agggagcaga accagcttat aagcggaggc gcggcaccaa
3121 gaagcgtgac caggaggagc atgccccttt ggcctatttg gcctggccca attcgcccta
3181 tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
3241 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
3301 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctatacg tacggcagtt
3361 taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg tacagagtga
3421 tattattgac acgccggggc gacggatggt gatccccctg gccagtgcac gtctgctgtc
3481 agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa gctggcgcat
3541 gatgaccacc gatatggcaa gtgtgccagt atccgttatc ggggaagaag tggctgatct
3601 cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg gaatataaat
3661 gtcaggcatg agattatcaa aaaggatctt cacctagatc cttttcacgt agaaagccag
3721 tccgcagaaa cggaccatga gtggacataa gcctcgttcg gttcgtaagc tgtaatgcaa
3781 gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg gtggtaacgg
3841 cgcagtggc
//
