| 质粒名称: | pSBbi-Pur |
|---|---|
| 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 |
|
| 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-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 |
买家导航
pSBbi-Pur载体 pSBbiPur质粒基本信息
pSBbi-Pur质粒图谱 pSBbiPur载体图谱和pSBbi-Pur载体序列 pSBbiPur质粒序列多克隆位点信息
pSBbi-Pur质粒 pSBbiPur载体简介
pSBbi-Pur质粒序列 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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