质粒名称: | pSBbi-GB |
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出品公司: | 爱迪基因 |
目录编号: | 60520 |
存储实验室: | Eric Kowarz |
载体骨架基本信息 |
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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: | 稻瘟 |
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-GB was a gift from Eric Kowarz (爱迪基因 plasmid # 60520) |
参考文献部分: | ptimized 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
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pSBbi-GB 6439 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: GFP and blasticidin resistance gene. ACCESSION . VERSION . KEYWORDS pSBbi-GB SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6439) 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-GB| FEATURES Location/Qualifiers source 1..6439 /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..3359 /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /translation="MLSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2688..2709) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2949..2968) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3296..3317 /label=EGFP-C /note="EGFP, forward primer" CDS 3369..3425 /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 3426..3824 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAMPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" primer_bind complement(3924..3943) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_site 3989..4044 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal" primer_bind complement(4043..4062) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(4436..4453) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4607..5195) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4687..4706) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5366..6226) /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 5989..6008 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6227..6331) /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 gagctgttca ccggggtggt gcccatcctg gtcgagctgg 2701 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct 2761 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca 2821 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga 2881 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct 2941 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc 3001 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc 3061 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga 3121 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg 3181 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc 3241 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg 3301 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg 3361 gcagtggagc tactaacttc agcctgctga agcaggctgg tgacgtcgag gagaatcctg 3421 gccccatggc catgcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta 3481 caatcaacag catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg 3541 acggccgcat cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac 3601 tcgtggtgct gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga 3661 tcggaaatga gaacaggggc atcttgagcc cctgcggacg gtgccgacag gtgcttctcg 3721 atctgcatcc tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg 3781 ggattcgtga attgctgccc tctggttatg tgtgggaggg ctaattcgaa ggcctgtcgt 3841 gaattcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc 3901 ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg 3961 aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg 4021 tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa 4081 catcagaatg agtatttggt ttagagtttg gcaacatatg ccatatgctg gctgccatga 4141 actagctact cgggacccct taccgaaaca tcgccgcatt ctgcagagga gtcgagtgta 4201 tgtaaacttc tgacccactg ggaatgtgat gaaagaaata aaagctgaaa tgaatcattc 4261 tctctactat tattctgata tttcacattc ttaaaataaa gtggtgatcc taactgacct 4321 aagacaggga atttttacta ggattaaatg tcaggaattg tgaaaaagtg agtttaaatg 4381 tatttggcta aggtgtatgt aaacttccga cttcaactgt atagggatcc gcttcctcgc 4441 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg 4501 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag 4561 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 4621 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 4681 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 4741 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 4801 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 4861 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 4921 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 4981 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 5041 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 5101 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca 5161 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 5221 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 5281 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 5341 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 5401 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 5461 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 5521 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 5581 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 5641 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 5701 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 5761 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 5821 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 5881 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 5941 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 6001 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 6061 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 6121 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 6181 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 6241 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 6301 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgatg 6361 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg 6421 ttaatatttt gttaaaatt //