| 质粒名称: | pCBC-DT1T2 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 50590 |
| 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: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | T1, U626t plus, U629p, T2 |
| 物种来源: | Synthetic |
| 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-DT1T2 was a gift from Qi-Jun Chen (Addgene plasmid # 50590 ; http://n2t.net/addgene:50590 ; RRID:Addgene_50590) |
| 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
pCBCDT1T2载体基本信息
pCBCDT1T2载体图谱质粒图谱和多克隆位点信息
pCBCDT1T2载体简介
pCBCDT1T2载体序列
LOCUS pCBC-DT1T2 3544 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-DT1T2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3544)
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 3544)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3544
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_RNA 20..95
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter complement(644..662)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(645..662)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(680..696)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(680..696)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(693..715)
/label=M13/pUC Reverse
/note="In lacZ gene"
promoter complement(735..782)
/label=EM7 promoter
/note="synthetic bacterial promoter "
primer_bind complement(793..816)
/label=pENTR-F
/note="pENTR vectors, forward primer"
terminator 824..851
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 943..1029
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
rep_origin complement(1194..1782)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1274..1293)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2007..2666)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKSKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 2581..2600
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
CDS complement(3015..3317)
/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 complement(3117..3136)
/label=ccdB-fwd
/note="ccdB gene, forward primer"
primer_bind 3440..3462
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 3454..3471
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 3455..3471
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3478..3497
/label=T7
/note="T7 promoter, forward primer"
promoter 3478..3496
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 gacggccatg gcggttctcg ttttagagct agaaatagca agttaaaata aggctagtcc
61 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tttgcaaaat tttccagatc
121 gatttcttct tcctctgttc ttcggcgttc aatttctggg gttttctctt cgttttctgt
181 aactgaaacc taaaatttga cctaaaaaaa atctcaaata atatgattca gtggttttgt
241 acttttcagt tagttgagtt ttgcagttcc gatgagataa accaatatta atccaaacta
301 ctgcagcctg acagacaaat gaggatgcaa acaattttaa agtttatcta acgctagctg
361 ttttgtttct tctctctggt gcaccaacga cggcgttttc tcaatcataa agaggcttgt
421 tttacttaag gccaataatg ttgatggatc gaaagaagag ggcttttaat aaacgagccc
481 gtttaagctg taaacgatgt caaaaacatc ccacatcgtt cagttgaaaa tagaagctct
541 gtttatatat tggtagagtc gactaagaga ttggcaaagt tatacaatgg cagtttagag
601 accaataatc cttggcctta agggccttgc atagcttgag tattctatag tgtcacctaa
661 atagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcggtt
721 tagttcctca ccttgtcgta ttatactatg ccgatatact atgccgatga ttaattgtca
781 acacttaagt aactaactaa caggaagagt ttgtagaaac gcaaaaaggc catccgtcag
841 gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc
901 tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag
961 agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg
1021 ttttatttga tgcctggcag ttccctactc tcgcttagta gttagacgtg cccgagatcc
1081 atcctagcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
1141 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
1201 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
1261 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
1321 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
1381 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
1441 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
1501 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
1561 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
1621 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
1681 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
1741 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
1801 tctacggggt ctgacgctca gtggaacggg gcgggcggcg ttgtgacaat ttaccgaaca
1861 actccgcggg acttgacctg atagtttggc tgtgagcaat tatgtgctta gtgcatctaa
1921 cgcttgagtt aagccgcgcc gcgaagcggc gtcggcttga acgaatttct agctagagga
1981 tcgcaccaat aactgcctta aaaaaattac gccccgccct gccactcatc gcagtactgt
2041 tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga
2101 atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat tgtgaaaacg
2161 ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag
2221 ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt
2281 tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg
2341 tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg
2401 tgaacactat cccatatcac cagctcaccg tctttcattg ccatacggaa ctccggatga
2461 gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttacttttc
2521 tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga
2581 gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg
2641 gtatatccag tgattttttt ctccatgatg tttaactttg ttttagggcg actgccctgc
2701 tgcgtaacat cgttgctgct ccataacatc aaacatcgac ccacggcgta acgcgcttgc
2761 tgcttggatg cccgaggcat agactgtacc ccaaaaaaac agtcataaca agccatgaaa
2821 accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgag
2881 cgcatacgct acttgcatta cagcttacga accgaacgag gcttatgtcc actcatggtc
2941 cgtttctgcg gactggcttt ctacgtgaaa aggatctagg tgaagatcct ttttgataat
3001 ctcatgcctg acatttatat tccccagaac atcaggttaa tggcgttttt gatgtcattt
3061 tcgcggtggc tgagatcagc cacttcttcc ccgataacgg atactggcac acttgccata
3121 tcggtggtca tcatgcgcca gctttcatcc ccgatatgca ccaccgggta aagttcacgg
3181 gagactttat ctgacagcag acgtgcactg gccaggggga tcaccatccg tcgccccggc
3241 gtgtcaataa tatcactctg tacatccaca aacagacgat aacggctctc tcttttatag
3301 gtgtaaacct taaactgccg tacgtatagg ctgcgcaact gttgggaagg gcgatcggtg
3361 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
3421 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa
3481 tacgactcac tatagggcga attgggccag gccaaatagg ccaaaggaat aatggtctct
3541 attg
//
