质粒名称: | pSBbi-BP |
---|---|
出品公司: | Addgene |
目录编号: | 60512 |
存储实验室: | Eric Kowarz |
载体骨架基本信息 |
|
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 |
|
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 |
|
材料和方法部分: | pSBbi-BP was a gift from Eric Kowarz (Addgene plasmid # 60512 ; http://n2t.net/addgene:60512 ; RRID:Addgene_60512) |
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
pSBbiBP载体基本信息
pSBbiBP载体图谱质粒图谱和多克隆位点信息
pSBbiBP载体简介
pSBbiBP载体序列
LOCUS pSBbi-BP 6622 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 puromycin resistance gene. ACCESSION . VERSION . KEYWORDS pSBbi-BP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6622) 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 6622) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6622 /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..4007 /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(3408..3427) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 3904..3924 /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind complement(4107..4126) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 4172..4227 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(4226..4245) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(4619..4636) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4790..5378) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4870..4889) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5549..6409) /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 6172..6191 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6410..6514) /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 accgagtaca 3421 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg 3481 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc 3541 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt 3601 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg 3661 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg 3721 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc 3781 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc 3841 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc 3901 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc 3961 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgattc gaaggcctgt 4021 cgtgaattca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat 4081 gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc 4141 atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata 4201 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta 4261 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgccatatg ctggctgcca 4321 tgaactagct actcgggacc ccttaccgaa acatcgccgc attctgcaga ggagtcgagt 4381 gtatgtaaac ttctgaccca ctgggaatgt gatgaaagaa ataaaagctg aaatgaatca 4441 ttctctctac tattattctg atatttcaca ttcttaaaat aaagtggtga tcctaactga 4501 cctaagacag ggaattttta ctaggattaa atgtcaggaa ttgtgaaaaa gtgagtttaa 4561 atgtatttgg ctaaggtgta tgtaaacttc cgacttcaac tgtataggga tccgcttcct 4621 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 4681 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 4741 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 4801 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 4861 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 4921 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 4981 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 5041 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 5101 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 5161 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 5221 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 5281 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 5341 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 5401 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 5461 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 5521 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 5581 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 5641 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 5701 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 5761 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 5821 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 5881 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 5941 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 6001 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 6061 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 6121 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 6181 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 6241 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 6301 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 6361 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 6421 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 6481 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 6541 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa 6601 gcgttaatat tttgttaaaa tt //
pSBbiBP载体图谱质粒图谱pdf版和相关资料下载
pSBbiBP载体质粒应用举例
- 上一篇:pCXLE-hOCT3/4
- 下一篇:pSBbi-Pur