质粒名称: | pCBASceI, pCBAScel |
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出品公司: | 爱迪基因 |
目录编号: | 26477 |
存储实验室: | Maria Jasin |
载体骨架基本信息 |
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载体名称: | pCAGGS |
载体出品公司: | -- |
载体抗性: | 氨苄青霉素 |
空载体大小: | 3073 bp |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 哺乳细胞表达 |
筛选标记: | -- |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | pCBASceI,CBAS |
物种来源: | -- |
插入大小: | 2583 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子 | -- |
标签/融合蛋白: | HA(插入件上的N端子) |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | XhoI(克隆过程中销毁) |
3' 克隆位点: | XhoI(克隆过程中销毁) |
5' 测序引物 | -- |
3' 测序引物 | -- |
文献引用方法 |
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材料和方法部分: | pCBASceI was a gift from Maria Jasin (Addgene plasmid # 26477 ; http://n2t.net/addgene:26477 ; RRID:Addgene_26477) |
参考文献部分: | Double-strand break repair by interchromosomal recombination: suppression of chromosomal translocations. Richardson C, Moynahan ME, Jasin M. Genes Dev. 1998 Dec 15. 12(24):3831-42. 10.1101/gad.12.24.3831 PubMed 9869637 |
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pCBAScel pCBASceI载体载体基本信息
pCBAScel pCBASceI载体载体图谱质粒图谱和多克隆位点信息
pCBAScel pCBASceI载体载体简介
pCBAScel pCBASceI载体载体序列
LOCUS pCBASceI 5665 bp ds-DNA circular SYN 19-APR-2019 DEFINITION I-SceI endonuclease expression vector with mammalian promoter to introduce a DSB at a genomic I-SceI site. ACCESSION . VERSION . KEYWORDS pCBASceI SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5665) AUTHORS Richardson C, Moynahan ME, Jasin M TITLE Double-strand break repair by interchromosomal recombination: suppression of chromosomal translocations. JOURNAL Genes Dev. 1998 Dec 15. 12(24):3831-42. PUBMED 9869637 REFERENCE 2 (bases 1 to 5665) AUTHORS . TITLE Direct Submission JOURNAL Exported Apr 19, 2019 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5665 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 5..384 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" intron 661..1677 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1601..1623 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1685..1704 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 1772..1792 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1814..1834 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1853..1879 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" primer_bind complement(2687..2706) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_site 2752..2807 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal" primer_bind complement(2806..2825) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(3168..3184) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3168..3184) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3181..3203) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3192..3208 /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(3216..3246) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3261..3282 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 3341..3536 /label=SV40 promoter /note="SV40 early promoter" rep_origin 3387..3522 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3449..3468 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 3542..3676 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3579..3598) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3633..3652 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3744..3761) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3915..4503) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3995..4014) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4674..5534) /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 5297..5316 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5535..5639) /gene="bla" /label=AmpR promoter ORIGIN 1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat 61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg 121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata 181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta 241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc 301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac 361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc 421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca 481 gcgatggggg cggggggggg ggggggcgcg ccaggcgggg gggggggggg gggggggggg 541 gggggggggg ggcggggggg ggcggcggca gccaatcaga gcggcgcgct ccgaaagttt 601 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg 661 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg 721 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct 781 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa 841 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc 901 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc 961 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg 1021 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg 1081 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac 1141 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt 1201 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg 1261 gggccgcctc gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc 1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg 1381 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc 1441 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag 1501 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg 1561 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg 1621 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc 1681 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt cctcgaaatt 1741 cagatccgcc gccactatgg gatcaagatc gccaaaaaag aagagaaagg tgccgaagaa 1801 gcatgcagca ccaccaaaaa aaaaacgaaa agtagaagac ccacgattta tgtacccata 1861 cgatgttcct gactatgcgg gtatgaaaaa catcaaaaaa aaccaggtaa tgaacctggg 1921 tccgaactct aaactgctga aagaatacaa atcccagctg atcgaactga acatcgaaca 1981 gttcgaagca ggtatcggtc tgatcctggg tgatgcttac atccgttctc gtgatgaagg 2041 taaaacctac tgtatgcagt tcgagtggaa aaacaaagca tacatggacc acgtatgtct 2101 gctgtacgat cagtgggtac tgtccccgcc gcacaaaaaa gaacgtgtta accacctggg 2161 taacctggta atcacctggg gcgcccagac tttcaaacac caagctttca acaaactggc 2221 taacctgttc atcgttaaca acaaaaaaac catcccgaac aacctggttg aaaactacct 2281 gaccccgatg tctctggcat actggttcat ggatgatggt ggtaaatggg attacaacaa 2341 aaactctacc aacaaatcga tcgtactgaa cacccagtct ttcactttcg aagaagtaga 2401 atacctggtt aagggtctgc gtaacaaatt ccaactgaac tgttacgtaa aaatcaacaa 2461 aaacaaaccg atcatctaca tcgattctat gtcttacctg atcttctaca acctgatcaa 2521 accgtacctg atcccgcaga tgatgtacaa actgccgaac actatctcct ccgaaacttt 2581 cctgaaataa taagtcgatc gaggaattca ctcctcaggt gcaggctgcc tatcagaagg 2641 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg 2701 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga 2761 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat 2821 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat 2881 atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga 2941 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt 3001 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg 3061 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc 3121 tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt 3181 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag 3241 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg 3301 cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa 3361 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 3421 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 3481 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc 3541 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 3601 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 3661 ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg 3721 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 3781 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 3841 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 3901 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 3961 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 4021 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 4081 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg 4141 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 4201 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 4261 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 4321 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg 4381 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 4441 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 4501 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 4561 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa 4621 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa 4681 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc 4741 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct 4801 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca 4861 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt 4921 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt 4981 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 5041 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc 5101 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 5161 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 5221 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 5281 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 5341 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 5401 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 5461 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 5521 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt 5581 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 5641 acatttcccc gaaaagtgcc acctg //
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