| 质粒名称: | pCBC-DT1T2.2 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 134752 |
| 存储实验室: | Qi-Jun Chen |
|
载体骨架基本信息 |
|
| 载体名称: | pCBC |
| 完整质粒大小: | 3364 bp |
| 载体类型: | CRISPR |
| 载体抗性: | Chloramphenicol |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: |
sgRNA-AtU6_29pgRNA
|
| 物种来源: | Synthetic |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | Unknown (unknown if destroyed) |
|
文献引用方法 |
|
| 材料和方法部分: | pCBC-DT1T2.2 was a gift from Qi-Jun Chen (Addgene plasmid # 134752 ; http://n2t.net/addgene:134752 ; RRID:Addgene_134752) |
| 参考文献部分: | A Novel Ternary Vector System United with Morphogenic Genes Enhances CRISPR/Cas Delivery in Maize. Zhang Q, Zhang Y, Lu MH, Chai YP, Jiang YY, Zhou Y, Wang XC, Chen QJ. Plant Physiol. 2019 Sep 26. pii: pp.19.00767. doi: 10.1104/pp.19.00767. 10.1104/pp.19.00767 PubMed 31558579 |
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-DT1T2.2 3364 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION PCR template for sgRNA cloning.
ACCESSION .
VERSION .
KEYWORDS pCBC-DT1T2.2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3364)
AUTHORS Zhang Q, Zhang Y, Lu MH, Chai YP, Jiang YY, Zhou Y, Wang XC, Chen QJ
TITLE A Novel Ternary Vector System United with Morphogenic Genes Enhances
CRISPR/Cas Delivery in Maize.
JOURNAL Plant Physiol. 2019 Sep 26. pii: pp.19.00767. doi:
10.1104/pp.19.00767.
PUBMED 31558579
REFERENCE 2 (bases 1 to 3364)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..3364
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..48
/label=EM7 promoter
/note="synthetic bacterial promoter "
primer_bind 68..90
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 87..103
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 87..103
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 121..139
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 121..138
/label=SP6
/note="SP6 promoter, forward primer"
misc_RNA 195..270
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
primer_bind complement(650..669)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(651..669)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(676..693)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(676..692)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(685..707)
/label=M13/pUC Forward
/note="In lacZ gene"
CDS 830..1132
/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 1011..1030
/label=ccdB-fwd
/note="ccdB gene, forward primer"
CDS 1481..2140
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKSKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(1547..1566)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
rep_origin 2365..2953
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2854..2873
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
terminator 3118..3204
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 3296..3323
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
primer_bind 3331..3354
/label=pENTR-F
/note="pENTR vectors, forward primer"
ORIGIN
1 gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac
61 taaaccgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
121 atttaggtga cactatagaa tactcaagct atgcaaggcc cttaaggcca aggtccactg
181 agcgtcaggt ctcagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat
241 caacttgaaa aagtggcacc gagtcggtgc ttttttttgc aaaattttcc agatcgattt
301 cttcttcctc tgttcttcgg cgttcaattt ctttaatcca aactactgca gcctgacaga
361 caaatgagga tgcaaacaat tttaaagttt atctaacgct agctgttttg tttcttctct
421 ctggtgcacc aacgacggcg ttttctcaat cataaagagg cttgttttac ttaaggccaa
481 taatgttgat ggatcgaaag aagagggctt ttaataaacg agcccgttta agctgtaaac
541 gatgtcaaaa acatcccaca tcgttcagtt gaaaatagaa gctctgttta tatattggta
601 gagtcgacta agagattggc ctttggccta tttggcctgg cccaattcgc cctatagtga
661 gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
721 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
781 ggcccgcacc gatcgccctt cccaacagtt gcgcagccta tacgtacggc agtttaaggt
841 ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat
901 tgacacgccg gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa
961 agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac
1021 caccgatatg gcaagtgtgc cagtatccgt tatcggggaa gaagtggctg atctcagcca
1081 ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg
1141 catgagatta tcaaaaagga tcttcaccta gatccttttc acgtagaaag ccagtccgca
1201 gaaacggacc atgagtggac ataagcctcg ttcggttcgt aagctgtaat gcaagtagcg
1261 tatgcgctca cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt
1321 ggcggttttc atggcttgtt atgactgttt ttttggggta cagtctatgc ctcgggcatc
1381 caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt
1441 tacgcagcag ggcagtcgcc ctaaaacaaa gttaaacatc atggagaaaa aaatcactgg
1501 atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc
1561 agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac
1621 cgtaaagaaa agtaagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat
1681 gaatgctcat ccggagttcc gtatggcaat gaaagacggt gagctggtga tatgggatag
1741 tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag
1801 tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta
1861 cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc
1921 caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt
1981 cgcccccgtt ttcacaatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct
2041 ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga
2101 attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg
2161 gtgcgatcct ctagctagaa attcgttcaa gccgacgccg cttcgcggcg cggcttaact
2221 caagcgttag atgcactaag cacataattg ctcacagcca aactatcagg tcaagtcccg
2281 cggagttgtt cggtaaattg tcacaacgcc gcccgccccg ttccactgag cgtcagaccc
2341 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
2401 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
2461 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
2521 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
2581 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
2641 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
2701 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
2761 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
2821 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
2881 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
2941 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
3001 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
3061 taggatggat ctcgggcacg tctaactact aagcgagagt agggaactgc caggcatcaa
3121 ataaaacgaa aggctcagtc ggaagactgg gcctttcgtt ttatctgttg tttgtcggtg
3181 aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgt gaagcaacgg
3241 cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaac taagcagaag
3301 gccatcctga cggatggcct ttttgcgttt ctacaaactc ttcctgttag ttagttactt
3361 aagt
//
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