质粒名称: | pMaCTag-P34 |
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出品公司: | Addgene |
目录编号: | 120030 |
存储实验室: | Michael Knop |
载体骨架基本信息 |
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载体名称: | pFA6a |
载体出品公司: | -- |
载体抗性: | 氨苄青霉素 |
空载体大小: | 2412 bp |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | PCR模板 |
筛选标记: | 嘌呤霉素 |
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: | 3xFLAG |
物种来源: | H. sapiens (human) |
插入大小: | -- |
基因突变: | -- |
GenBank编号 | -- |
启动子 | -- |
标签/融合蛋白: | 3xFLAG(主干上的C终端) |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | BamHI(未破坏) |
3' 克隆位点: | SpeI (未破坏) |
5' 测序引物 | SP6 |
3' 测序引物 | AAACCACAACTAGAATGCAGTG |
文献引用方法 |
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材料和方法部分: | pMaCTag-P34 was a gift from Michael Knop (Addgene plasmid # 120030 ; http://n2t.net/addgene:120030 ; RRID:Addgene_120030) |
参考文献部分: | CRISPR-Cas12a-assisted PCR tagging of mammalian genes. Fueller J, Herbst K, Meurer M, Gubicza K, Kurtulmus B, Knopf JD, Kirrmaier D, Buchmuller BC, Pereira G, Lemberg MK, Knop M. J Cell Biol. 2020 Jun 1;219(6). pii: 151766. doi: 10.1083/jcb.201910210. 10.1083/jcb.201910210 PubMed 32406907 |
买家导航
pMaCTagP34载体基本信息
pMaCTagP34载体图谱质粒图谱和多克隆位点信息
pMaCTagP34载体简介
pMaCTagP34载体序列
LOCUS pMaCTag-P34 4159 bp ds-DNA circular SYN 08-JAN-2019 DEFINITION Template for C-terminal PCR tagging of mammalian genes (Puromycin selection) with 3xFLAG. ACCESSION . VERSION . KEYWORDS pMaCTag-P34 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4159) AUTHORS Fueller J, Meurer M, Herbst K, Gubicza K, Kurtulmus B, Knopf JD, Kirrmaier D, Buchmuller B, Pereira G, Lemberg MK, Knop M TITLE CRISPR/Cas12a-assisted PCR tagging of mammalian genes JOURNAL bioRxiv REFERENCE 2 (bases 1 to 4159) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 8, 2019 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4159 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..19 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 1..18 /label=SP6 /note="SP6 promoter, forward primer" CDS 81..146 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" polyA_signal 156..277 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(193..212) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 247..266 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" protein_bind 284..317 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." promoter 324..640 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 491..626 /label=SV40 ori /note="SV40 origin of replication" primer_bind 553..572 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 1262..1486 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" promoter 1493..1733 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 1493..1513 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 1664..1683 /label=LKO.1 5' /note="Human U6 promoter, forward primer" protein_bind 1743..1776 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind complement(1813..1832) /label=T7 /note="T7 promoter, forward primer" promoter complement(1814..1832) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1919..1936) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2090..2678) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2170..2189) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2849..3709) /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 3472..3491 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3710..3814) /gene="bla" /label=AmpR promoter primer_bind 3882..3900 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(3938..3960) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 4060..4079 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 atttaggtga cactatagaa cgcggccgcc agctgaagct tcgtacgtca ggtggaggag 61 gtagtggcgg aggcggatcc gactacaaag accatgacgg tgattataaa gatcatgata 121 tcgattacaa ggatgacgat gacaagtaga ctagtaactt gtttattgca gcttataatg 181 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt 241 ctagttgtgg tttgtccaaa ctcatcaatg tatcttagtc gatataactt cgtatagcat 301 acattatacg aagttatgtc gacggtgtgg aaagtcccca ggctccccag caggcagaag 361 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc 421 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct 481 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg 541 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa 601 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gaattcgcca ccatgactga 661 gtacaagcct actgtgcgtt tagccaccag ggatgatgtg ccacgagccg tgaggacact 721 agcagccgcc tttgctgact acccagctac aaggcacacc gtggaccctg acagacacat 781 tgaaagagtc acagaacttc aggagctgtt tttaactagg gtgggcctgg atattggcaa 841 agtctgggtg gccgatgatg gagcagcagt tgcagtgtgg accaccccag agtctgttga 901 agctggagct gtctttgcag aaattggacc aagaatggcc gagttatctg gcagcaggct 961 ggctgctcaa cagcagatgg aaggattatt agctccccac agacccaagg agcctgcctg 1021 gttcctggca acagttggag tttctccaga ccatcagggc aagggcttag gatctgctgt 1081 ggtgctgcct ggtgtggaag cagctgagag agctggggtg ccagcatttc tggagacatc 1141 agctcctaga aacctgccct tctatgaacg gctgggcttc acagtgactg ctgatgtgga 1201 ggtgcctgag ggccccagaa cctggtgcat gacaagaaaa ccaggagcct gactgcagcg 1261 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc 1321 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt 1381 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat 1441 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggctcg aggagggcct 1501 atttcccatg attccttcat atttgcatat acgatacaag gctgttagag agataattgg 1561 aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata 1621 atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac 1681 cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaacac 1741 cgataacttc gtatagcata cattatacga agttatggta ccgatgcagc tagcccgcgg 1801 atctgccggt ctccctatag tgagtcgtat taatttcgat aagccaggtt aacctgcatt 1861 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 1921 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 1981 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 2041 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 2101 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 2161 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 2221 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 2281 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 2341 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 2401 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 2461 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 2521 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 2581 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 2641 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 2701 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 2761 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 2821 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 2881 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 2941 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 3001 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 3061 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 3121 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 3181 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 3241 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 3301 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 3361 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 3421 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 3481 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 3541 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 3601 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 3661 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 3721 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 3781 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 3841 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc 3901 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg 3961 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt 4021 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac 4081 tgagagtgca ccatatggac atattgtcgt tagaacgcgg ctacaattaa tacataacct 4141 tatgtatcat acacatacg //
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