质粒名称: | pSBbi-Pur |
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出品公司: | Addgene |
目录编号: | 60523 |
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: | 嘌呤霉素 |
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-Pur was a gift from Eric Kowarz (Addgene plasmid # 60523 ; http://n2t.net/addgene:60523 ; RRID:Addgene_60523) |
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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pSBbiPur载体基本信息
pSBbiPur载体图谱质粒图谱和多克隆位点信息
pSBbiPur载体简介
pSBbiPur载体序列
LOCUS pSBbi-Pur 5857 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: puromycin resistance gene. ACCESSION . VERSION . KEYWORDS pSBbi-Pur SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5857) 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 5857) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5857 /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..3242 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(2643..2662) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 3139..3159 /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind complement(3342..3361) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 3407..3462 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(3461..3480) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(3854..3871) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4025..4613) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4105..4124) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4784..5644) /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 5407..5426 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5645..5749) /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 ttagtgagaa 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 gcccaagcca ggcctcaact 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 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg 2701 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg 2761 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg 2821 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg 2881 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg 2941 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg 3001 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg 3061 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc 3121 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg 3181 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct 3241 gattcgaagg cctgtcgtga attcactcct caggtgcagg ctgcctatca gaaggtggtg 3301 gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa 3361 aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt 3421 attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg 3481 agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc 3541 atatgctggc tgccatgaac tagctactcg ggacccctta ccgaaacatc gccgcattct 3601 gcagaggagt cgagtgtatg taaacttctg acccactggg aatgtgatga aagaaataaa 3661 agctgaaatg aatcattctc tctactatta ttctgatatt tcacattctt aaaataaagt 3721 ggtgatccta actgacctaa gacagggaat ttttactagg attaaatgtc aggaattgtg 3781 aaaaagtgag tttaaatgta tttggctaag gtgtatgtaa acttccgact tcaactgtat 3841 agggatccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 3901 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 3961 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 4021 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 4081 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 4141 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 4201 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 4261 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 4321 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 4381 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 4441 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 4501 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 4561 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 4621 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 4681 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 4741 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 4801 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 4861 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 4921 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 4981 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 5041 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 5101 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 5161 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 5221 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 5281 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 5341 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 5401 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 5461 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 5521 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 5581 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 5641 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 5701 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 5761 gaaaagtgcc acctgatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc 5821 atcaggaaat tgtaagcgtt aatattttgt taaaatt //
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