质粒名称: | pSBbi-BH |
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出品公司: | Addgene |
目录编号: | 60515 |
存储实验室: | Eric Kowarz |
载体骨架基本信息 |
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载体名称: | pUC19 |
空载体大小: | 2021 bp |
载体类型: | 哺乳动物表达;转座子 |
筛选标记: | 潮霉素 |
载体抗性: | 氨苄霉素 |
生长温度: | 37 ℃ |
宿主菌: | Top10 |
拷贝数: | 高拷贝 |
插入基因信息 |
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启动子: | EF1a/RPBSA |
克隆方法: | 限制性内切酶 |
5’测序引物 | GCCTCAGACAGTGGTTCAAAG |
3’测序引物 | AGGCACAGTCGAGGCTGAT |
文献引用方法 |
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材料和方法部分: | pSBbi-BH was a gift from Eric Kowarz (Addgene plasmid # 60515 ; http://n2t.net/addgene:60515 ; RRID:Addgene_60515) |
参考文献部分: | Optimized Sleeping Beauty transposons rapidly generate stable transgenic cell lines. Kowarz E, Loescher D, Marschalek R. Biotechnol J. 2015 Feb 4. doi: 10.1002/biot.201400821. 10.1002/biot.201400821 PubMed 25650551 |
买家导航
pSBbiBH载体基本信息
pSBbiBH载体图谱质粒图谱和多克隆位点信息
pSBbiBH载体简介
pSBbiBH载体序列
LOCUS pSBbi-BH 7048 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION SB-transposon with constitutive bi-directional promoter, one side: SfiI cloning site for GOI (contains filler DNA), other side: BFP and hygromycin resistance gene. ACCESSION . VERSION . KEYWORDS pSBbi-BH SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7048) AUTHORS Kowarz E, Loescher D, Marschalek R TITLE Optimized Sleeping Beauty transposons rapidly generate stable transgenic cell lines. JOURNAL Biotechnol J. 2015 Feb 4. doi: 10.1002/biot.201400821. PUBMED 25650551 REFERENCE 2 (bases 1 to 7048) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7048 /organism="synthetic DNA construct" /mol_type="other DNA" polyA_signal 324..548 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind 537..554 /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" regulatory 794..803 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" promoter complement(829..2010) /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" primer_bind complement(861..881) /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS 3351..3407 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 3408..4433 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" primer_bind complement(4533..4552) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 4598..4653 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(4652..4671) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(5045..5062) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5216..5804) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5296..5315) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5975..6835) /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 6598..6617 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6836..6940) /gene="bla" /label=AmpR promoter ORIGIN 1 ggatccctat acagttgaag tcggaagttt acatacactt aagttggagt cattaaaact 61 cgtttttcaa ctactccaca aatttcttgt taacaaacaa tagttttggc aagtcagtta 121 ggacatctac tttgtgcatg acacaagtca tttttccaac aattgtttac agacagatta 181 tttcacttat aattcactgt atcacaattc cagtgggtca gaagtttaca tacactaagt 241 tcgactcctc tgcagaatgc ggcgatgttt cggtaagggg tccgctatct agtaggccca 301 gctggttctt tccgcctcag aagccataga gcccaccgca tccccagcat gcctgctatt 361 gtcttcccaa tcctccccct tgctgtcctg ccccacccca ccccccagaa tagaatgaca 421 cctactcaga caatgcgatg caatttcctc attttattag gaaaggacag tgggagtggc 481 accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct ggcaactaga 541 aggcacagtc gaggctgatc agcgagctct agagaattga tccccaagct tggcctgaca 601 ggccttcgaa ttagagagaa ggaaagacgc aaccaggaag cccaccaaga gaaagaggca 661 tttcctgaaa agattcccct ttttggagag ccctacaagg tatttactga acactagaca 721 ttgaagtcct gatttatcac aatgttgaac cctatgatga aacaattcag tataattgag 781 ttcgaaactc gagccatggt ggcctcagag gccagcttgg ggtagttttc acgacacctt 841 aaatggaaga aaaaaacttt gaaccactgt ctgaggcttg agaatgaacc aagatccaaa 901 ctcaaaaagg gcaaattcca aggagaatta catcaagtgc caagctggcc taacttcagt 961 ctccacccac tcagtatggg gaaactccat cgcataaaac ccctcccccc aacctaaaga 1021 cgacgtactc caaaagctcg agaactaatc gaggtgcctg gacggcgccc ggtactccgt 1081 ggagtcacat gaagcgacgg ctgaggacgg aaaggccctt ttcctttgtg tgggtgactc 1141 acccgcccgc tctcccgagc gccgcgtcct ccattttgag ctccctgcag cagggccggg 1201 aagcggccat ctttccgctc acgcaactgg tgccgaccgg gccagccttg ccgcccaggg 1261 cggggcgata cacggcggcg cgaggccagg caccagagca ggccggccag cttgagacta 1321 cccccgtccg attctcggtg gccgcgctcg caggccccgc ctcgccgaac atgtgcgctg 1381 ggacgcacgg gccccgtcgc cgcccgcggc cccaaaaacc gaaataccag tgtgcagatc 1441 ttggcccgca tttacaagac tatcttgcca gaaaaaaagc gtcgcagcag gtcatcaaaa 1501 attttaaatg gctagagact tatcgaaagc agcgagacag gcgcgaaggt gccaccagat 1561 tcgcacgcgg cggccccagc gcccaggcca agcctcaact caagcacgag gcgaaggggc 1621 tccttaagcg caaggcctcg aactctccca cccacttcca acccgaagct cgggatcaag 1681 aatcacgtac tgcagccagg ggcgtggaag taattcaagg cacgcaaggg ccataacccg 1741 taaagaggcc aggcccgcgg gaaccacaca cggcacttac ctgtgttctg gcggcaaacc 1801 cgttgcgaaa aagaacgttc acggcgacta ctgcacttat atacggttct cccccaccct 1861 cgggaaaaag gcggagccag tacacgacat cactttccca gtttaccccg cgccaccttc 1921 tctaggcacc ggttcaattg ccgacccctc cccccaactt ctcggggact gtgggcgatg 1981 tgcgctctgc ccactgacgg gcaccggagc ctctaggcga gaccctgtct cacaaaataa 2041 agtaagcccg gactgagtgc ggaaaggcgg gcctggcggg tctggtctcc ccatgcgggc 2101 caccagaggc cctgcagcct tcagtcgctt gaaggggtaa tggcgcttcc actcacaaac 2161 atggcggaca gagcgtgtga acgagatgaa cagcccctca aaaatatggc cgccgaggct 2221 ggacggccgt gccccagcag caccgcctcc gcgccccacg tgatctctcg ccgggcacag 2281 cgctgaccgc ggaggtccaa ccggaagaat gtccggattg gacattcgga agagggcccg 2341 ccttccctgg ggaatctctg cgcacgcgca gaacgcttcg accaatgaaa acacaggaag 2401 ccgtccgcgc aaccgcgttg cgtcacttct gccgcccctg tttcaaggta tatagccgta 2461 gacggaactt cgcctttctc tcggccttag cgccattttt ttgggtgagt gttttttggt 2521 tcctgcgttg ggattccgtg tacaatccat agacatctga cctcggcact tagcatcatc 2581 acagcaaact aactgtagcc tttctctctt tccctgtaga aacctctgca cctgaggcca 2641 ccatgagcga gctgattaag gagaacatgc acatgaagct gtacatggag ggcaccgtgg 2701 acaaccatca cttcaagtgc acatccgagg gcgaaggcaa gccctacgag ggcacccaga 2761 ccatgagaat caaggtggtc gagggcggcc ctctcccctt cgccttcgac atcctggcta 2821 ctagcttcct ctacggcagc aagaccttca tcaaccacac ccagggcatc cccgacttct 2881 tcaagcagtc cttccctgag ggcttcacat gggagagagt caccacatac gaagacgggg 2941 gcgtgctgac cgctacccag gacaccagcc tccaggacgg ctgcctcatc tacaacgtca 3001 agatcagagg ggtgaacttc acatccaacg gccctgtgat gcagaagaaa acactcggct 3061 gggaggcctt caccgagacg ctgtaccccg ctgacggcgg cctggaaggc agaaacgaca 3121 tggccctgaa gctcgtgggc gggagccatc tgatcgcaaa catcaagacc acatatagat 3181 ccaagaaacc cgctaagaac ctcaagatgc ctggcgtcta ctatgtggac tacagactgg 3241 aaagaatcaa ggaggccaac aacgagacct acgtcgagca gcacgaggtg gcagtggcca 3301 gatactgcga cctccctagc aaactggggc acaagctgaa cggcagtgga gctactaact 3361 tcagcctgct gaagcaggct ggtgacgtcg aggagaatcc tggccccatg aaaaagcctg 3421 aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc 3481 tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg 3541 gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc 3601 ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattcagcg 3661 agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg 3721 aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg 3781 ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca 3841 ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg 3901 tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg 3961 ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc 4021 tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt 4081 cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc 4141 agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt 4201 atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg 4261 atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg 4321 ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac 4381 tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa tagttcgaag 4441 gcctgtcgtg aattcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg 4501 gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg 4561 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt 4621 gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat 4681 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc catatgctgg 4741 ctgccatgaa ctagctactc gggacccctt accgaaacat cgccgcattc tgcagaggag 4801 tcgagtgtat gtaaacttct gacccactgg gaatgtgatg aaagaaataa aagctgaaat 4861 gaatcattct ctctactatt attctgatat ttcacattct taaaataaag tggtgatcct 4921 aactgaccta agacagggaa tttttactag gattaaatgt caggaattgt gaaaaagtga 4981 gtttaaatgt atttggctaa ggtgtatgta aacttccgac ttcaactgta tagggatccg 5041 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 5101 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 5161 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 5221 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 5281 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 5341 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 5401 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 5461 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 5521 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 5581 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 5641 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 5701 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 5761 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 5821 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 5881 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 5941 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 6001 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 6061 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 6121 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 6181 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 6241 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 6301 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 6361 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 6421 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 6481 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 6541 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 6601 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 6661 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 6721 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 6781 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 6841 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 6901 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 6961 cacctgatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa 7021 ttgtaagcgt taatattttg ttaaaatt //
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