质粒名称: | pCBC-MT1T2.2 |
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出品公司: | Addgene |
目录编号: | 134751 |
存储实验室: | Qi-Jun Chen |
载体骨架基本信息 |
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载体名称: | pCBC |
完整质粒大小: | 3605bp |
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 |
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插入基因名称: | sgRNA-TaU3p |
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 |
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材料和方法部分: | pCBC-MT1T2.2 was a gift from Qi-Jun Chen (Addgene plasmid # 134751 ; http://n2t.net/addgene:134751 ; RRID:Addgene_134751) |
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 |
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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-MT1T2.2 3605 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION PCR template for sgRNA cloning. ACCESSION . VERSION . KEYWORDS pCBC-MT1T2.2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3605) 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 3605) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..3605 /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 194..269 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" primer_bind complement(891..910) /label=T7 /note="T7 promoter, forward primer" promoter complement(892..910) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(917..934) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(917..933) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(926..948) /label=M13/pUC Forward /note="In lacZ gene" CDS 1071..1373 /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 1252..1271 /label=ccdB-fwd /note="ccdB gene, forward primer" CDS 1722..2381 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKSKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(1788..1807) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" rep_origin 2606..3194 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3095..3114 /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 3359..3445 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 3537..3564 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" primer_bind 3572..3595 /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 aggggcttcg 181 tgccaaccaa cgagttttag agctagaaat agcaagttaa aataaggcta gtccgttatc 241 aacttgaaaa agtggcaccg agtcggtgct tttttttttc gttttgcatt gagttttctc 301 cgtcgcatgt ttgcagcatg aatccaaacc acacggagtt caaattccca cagattaagg 361 ctcgtccgtc gcacaaggta atgtgtgaat attatatctg tcgtgcaaaa ttgcctggcc 421 tgcacaattg ctgttatagt tggcggcagg gagagtttta acattgacta gcgtgctgat 481 aatttgtgag aaataataat tgacaagtag atactgacat ttgagaagag cttctgaact 541 gttattagta acaaaaatgg aaagctgatg cacggaaaaa ggaaagaaaa agccatactt 601 ttttttaggt aggaaaagaa aaagccatac gagactgatg tctctcagat gggccgggat 661 ctgtctatct agcaggcagc agcccaccaa cctcacgggc cagcaattac gagtccttct 721 aaaagctccc gccgaggggc gctggcgctg ctgtgcagca gcacgtctaa cattagtccc 781 acctcgccag tttacaggga gcagaaccag cttataagcg gaggcgcggc accaagaagc 841 gtgaccagga ggagcatgcc cctttggcct atttggcctg gcccaattcg ccctatagtg 901 agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg 961 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag 1021 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct atacgtacgg cagtttaagg 1081 tttacaccta taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta 1141 ttgacacgcc ggggcgacgg atggtgatcc ccctggccag tgcacgtctg ctgtcagata 1201 aagtctcccg tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga 1261 ccaccgatat ggcaagtgtg ccagtatccg ttatcgggga agaagtggct gatctcagcc 1321 accgcgaaaa tgacatcaaa aacgccatta acctgatgtt ctggggaata taaatgtcag 1381 gcatgagatt atcaaaaagg atcttcacct agatcctttt cacgtagaaa gccagtccgc 1441 agaaacggac catgagtgga cataagcctc gttcggttcg taagctgtaa tgcaagtagc 1501 gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag 1561 tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg cctcgggcat 1621 ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg 1681 ttacgcagca gggcagtcgc cctaaaacaa agttaaacat catggagaaa aaaatcactg 1741 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt 1801 cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga 1861 ccgtaaagaa aagtaagcac aagttttatc cggcctttat tcacattctt gcccgcctga 1921 tgaatgctca tccggagttc cgtatggcaa tgaaagacgg tgagctggtg atatgggata 1981 gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga 2041 gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt 2101 acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag 2161 ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct 2221 tcgcccccgt tttcacaatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc 2281 tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg 2341 aattacaaca gtactgcgat gagtggcagg gcggggcgta atttttttaa ggcagttatt 2401 ggtgcgatcc tctagctaga aattcgttca agccgacgcc gcttcgcggc gcggcttaac 2461 tcaagcgtta gatgcactaa gcacataatt gctcacagcc aaactatcag gtcaagtccc 2521 gcggagttgt tcggtaaatt gtcacaacgc cgcccgcccc gttccactga gcgtcagacc 2581 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 2641 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 2701 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag 2761 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 2821 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 2881 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 2941 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 3001 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 3061 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 3121 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 3181 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc 3241 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg 3301 ctaggatgga tctcgggcac gtctaactac taagcgagag tagggaactg ccaggcatca 3361 aataaaacga aaggctcagt cggaagactg ggcctttcgt tttatctgtt gtttgtcggt 3421 gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg tgaagcaacg 3481 gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ctaagcagaa 3541 ggccatcctg acggatggcc tttttgcgtt tctacaaact cttcctgtta gttagttact 3601 taagt //
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