质粒名称: | pFUS_B6 |
---|---|
出品公司: | 爱迪基因 |
目录编号: | 31023 |
存储实验室: | Adam Bogdanove, Daniel Voytas |
载体骨架基本信息 |
|
载体名称: | pCR8 |
载体出品公司: | Invitrogen |
载体抗性: | 大观霉素 |
空载体大小: | 2447 bp |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
|
插入基因名称: | LacZ + BsaI restriction sites |
插入大小: | 459 bp |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | AflII (未破坏) |
3' 克隆位点: | XbaI(未破坏) |
5' 测序引物 | pCR8_F1 (ttgatgcctggcagttccct) |
3' 测序引物 | pCR8_R1 (cgaaccgaacaggcttatgt) |
文献引用方法 |
|
材料和方法部分: | pFUS_B6 was a gift from Adam Bogdanove & Daniel Voytas (Addgene plasmid # 31023 ; http://n2t.net/addgene:31023 ; RRID:Addgene_31023) |
参考文献部分: | Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting. Cermak T, Doyle EL, Christian M, Wang L, Zhang Y, Schmidt C, Baller JA, Somia NV, Bogdanove AJ, Voytas DF. Nucleic Acids Res. 2011 Apr 14. ():. 10.1093/nar/gkr218 PubMed 21493687 |
买家导航
pFUSB6载体基本信息
pFUSB6载体图谱质粒图谱和多克隆位点信息
pFUSB6载体简介
pFUSB6载体序列
LOCUS pFUS_B6 3019 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pFUS_B6 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3019) AUTHORS Cermak T, Doyle EL, Christian M, Wang L, Zhang Y, Schmidt C, Baller JA, Somia NV, Bogdanove AJ, Voytas DF TITLE Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting. JOURNAL Nucleic Acids Res. 2011 Apr 14. ():. PUBMED 21493687 REFERENCE 2 (bases 1 to 3019) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3019 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 67..83 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 295..311 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 321..340 /label=T7 /note="T7 promoter, forward primer" promoter 321..339 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(387..407) /label=T3 /note="T3 promoter, forward primer" promoter complement(387..405) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(426..442) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(426..442) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(439..461) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 450..466 /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(474..504) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 519..540 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." CDS 1015..1806 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" rep_origin 1899..2487 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2388..2407 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2641..2658 /label=L4440 /note="L4440 vector, forward primer" terminator 2817..2844 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 2936..3 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" CDS complement(2943..430) /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLDSH HLVRGQAMRRLRLAVVLQRRDWENIHASVNARVGNCQASNKTKGSVGRLGLSFYLLFVG ERSPE" ORIGIN 1 ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac 61 gacgttgtaa aacgacggcc agtcttaagc gtctcatggc ctgaccccgg accaagtggt 121 ggctatcgag accggcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact 181 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat 241 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa 301 cgacggccag tgagcgcgcg taatacgact cactataggg cgaattgggt accgggcccc 361 ccctcgaggt cctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa 421 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 481 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 541 attgcgttgc gctcactgcc cgctttccac cggtggtctc acggtgcagc ggctgttgcc 601 ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg gtggctatcg agacgtctag 661 accagccagg acagaaatgc ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca 721 ccgtggaaac ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg 781 taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt 841 ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct 901 atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg 961 agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa cattatgagg 1021 gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc 1081 catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg 1141 aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg 1201 cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt 1261 ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca 1321 gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc 1381 gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc 1441 gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta 1501 tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat 1561 gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc 1621 gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc 1681 gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc 1741 gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc 1801 aaataaccct cgagccaccc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac 1861 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc 1921 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat 1981 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat 2041 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct 2101 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt 2161 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg 2221 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta 2281 cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg 2341 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg 2401 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc 2461 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg 2521 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat 2581 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc 2641 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg 2701 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt 2761 gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa 2821 ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc 2881 ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg 2941 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg 3001 actgagcctt tcgttttat //