质粒名称: | pSBbi-RN |
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出品公司: | 爱迪基因 |
目录编号: | 60519 |
存储实验室: | Eric Kowarz |
载体骨架基本信息 |
|
载体名称: | pUC19 |
载体抗性: | 氨苄霉素 |
空载体大小: | 2021 bp |
载体类型: | 哺乳细胞表达,Transposon |
筛选标记: | 新霉素(G418) |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
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-RN was a gift from Eric Kowarz (爱迪基因 plasmid # 60519) |
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 |
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载体基本信息
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载体简介
载体序列
LOCUS pSBbi-RN 6820 bp ds-DNA circular SYN 10-JAN-2018 DEFINITION SB-transposon with constitutive bi-directional promoter, one side: SfiI cloning site for GOI (contains filler DNA), other side: RFP and neomycin resistance gene. ACCESSION . VERSION . KEYWORDS pSBbi-RN SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6820) 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 COMMENT This file is created by Vector NTI http://www.biofeng.com/ COMMENT ORIGDB|GenBank COMMENT VNTAUTHORNAME|biofeng.com| COMMENT VNTNAME|pSBbi-RN| FEATURES Location/Qualifiers source 1..6820 /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 2643..3344 /codon_start=1 /product="dimeric variant of DsRed fluorescent protein (Shaner et al., 2004)" /label=dTomato /note="mammalian codon-optimized" /translation="MLSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH HLFLYGMDELYK" primer_bind complement(2776..2794) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 3044..3063 /label=mCherry-F /note="mCherry, forward primer" primer_bind 3238..3261 /label=DsRed1-C /note="DsRed1, forward primer" CDS 3354..3410 /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 3411..4205 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3465..3484) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4075..4094 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4305..4324) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_site 4370..4425 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal" primer_bind complement(4424..4443) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(4817..4834) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4988..5576) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5068..5087) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5747..6607) /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 6370..6389 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6608..6712) /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 ccatgttgag caagggcgag gaggtcatca aagagttcat gcgcttcaag gtgcgcatgg 2701 agggctccat gaacggccat gagttcgaga tcgagggcga gggcgagggc cgcccctacg 2761 agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg 2821 acatcctgtc cccccagttc atgtacggct ccaaggcgta cgtgaagcac cccgccgaca 2881 tccccgatta caagaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact 2941 tcgaggacgg cggtctggtg accgtgaccc aggactcctc cctgcaggac ggcacgctga 3001 tctacaaggt gaagatgcgc ggcaccaact tcccccccga cggccccgta atgcagaaga 3061 agacaatggg ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg 3121 gcgagatcca ccaggccctg aagctgaagg acggcggcca ctacctggtg gagttcaaga 3181 ccatctacat ggccaagaag cccgtgcaac tgcccggcta ctactacgtg gacaccaagc 3241 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag cgctccgagg 3301 gccgccacca cctgttcctg tacggcatgg acgagctgta caagggcagt ggagctacta 3361 acttcagcct gctgaagcag gctggtgacg tcgaggagaa tcctggcccc atgattgaac 3421 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact 3481 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc 3541 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg 3601 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg 3661 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt 3721 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc 3781 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag 3841 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg 3901 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc 3961 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt 4021 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg 4081 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt 4141 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct 4201 tctgattcga aggcctgtcg tgaattcact cctcaggtgc aggctgccta tcagaaggtg 4261 gtggctggtg tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc 4321 aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa 4381 tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat 4441 gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat 4501 gccatatgct ggctgccatg aactagctac tcgggacccc ttaccgaaac atcgccgcat 4561 tctgcagagg agtcgagtgt atgtaaactt ctgacccact gggaatgtga tgaaagaaat 4621 aaaagctgaa atgaatcatt ctctctacta ttattctgat atttcacatt cttaaaataa 4681 agtggtgatc ctaactgacc taagacaggg aatttttact aggattaaat gtcaggaatt 4741 gtgaaaaagt gagtttaaat gtatttggct aaggtgtatg taaacttccg acttcaactg 4801 tatagggatc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 4861 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 4921 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 4981 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 5041 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 5101 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 5161 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 5221 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 5281 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 5341 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 5401 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc 5461 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 5521 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 5581 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 5641 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 5701 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 5761 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 5821 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 5881 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 5941 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 6001 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 6061 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 6121 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 6181 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 6241 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 6301 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 6361 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 6421 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 6481 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 6541 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 6601 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 6661 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 6721 cccgaaaagt gccacctgat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac 6781 cgcatcagga aattgtaagc gttaatattt tgttaaaatt //
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