| 质粒名称: | pBABE-puro-GFP-wt-lamin A |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 17662 |
| 存储实验室: | Tom Misteli |
|
载体骨架基本信息 |
|
| 载体名称: | pBABE-puro |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5200 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达,逆转录病毒 |
| 筛选标记: | 嘌呤霉素 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | GFP-lamin A，lamin A |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | EGFP（插入端N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamHI(克隆过程中销毁) |
| 3' 克隆位点: | BamHI(克隆过程中销毁) |
| 5' 测序引物 | EGFP-C |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pBABE-puro-GFP-wt-lamin A was a gift from Tom Misteli (Addgene plasmid # 17662 ; http://n2t.net/addgene:17662 ; RRID:Addgene_17662) |
| 参考文献部分: | Lamin A-dependent misregulation of adult stem cells associated with accelerated ageing. Scaffidi P, Misteli T. Nat Cell Biol. 2008 Mar 2. ():. 10.1038/ncb1708 PubMed 18311132 |
买家导航
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 pBABE-puro-GFP-wt-lamin A 8170 bp ds-DNA circular SYN 19-MAR-2019
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pBABE-puro-GFP-wt-lamin A
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8170)
AUTHORS Scaffidi P, Misteli T
TITLE Lamin A-dependent misregulation of adult stem cells associated with
accelerated ageing.
JOURNAL Nat Cell Biol. 2008 Mar 2. ():.
PUBMED 18311132
REFERENCE 2 (bases 1 to 8170)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Mar 19, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8170
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 130..546
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 476..498
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 514..530
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
regulatory 587..596
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 593..1309
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(638..659)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(899..918)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1246..1267
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(3688..3708)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3693..4022
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3873..4008
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3935..3954
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4032..4631
/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(4032..4051)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 4528..4548
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 4852..5321
/note="long terminal repeat from Moloney murine leukemia
virus"
primer_bind complement(5451..5468)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5622..6210)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5702..5721)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6370..7230)
/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 6993..7012
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7231..7335)
/gene="bla"
/label=AmpR promoter
LTR 7366..7835
/note="long terminal repeat from Moloney murine leukemia
virus"
misc_feature 7900..8099
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
ORIGIN
1 ttcccgcctc cgtctgaatt tttgctttcg gtttggaacc gaagccgcgc gtcttgtctg
61 ctgcagcgct gcagcatcgt tctgtgttgt ctctgtctga ctgtgtttct gtatttgtct
121 gaaaattagg gccagactgt taccactccc ttaagtttga ccttaggtca ctggaaagat
181 gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga agagacgttg ggttaccttc
241 tgctctgcag aatggccaac ctttaacgtc ggatggccgc gagacggcac ctttaaccga
301 gacctcatca cccaggttaa gatcaaggtc ttttcacctg gcccgcatgg acacccagac
361 caggtcccct acatcgtgac ctgggaagcc ttggcttttg acccccctcc ctgggtcaag
421 ccctttgtac accctaagcc tccgcctcct cttcctccat ccgccccgtc tctccccctt
481 gaacctcctc gttcgacccc gcctcgatcc tccctttatc cagccctcac tccttctcta
541 ggcgccggcc ggatcccagt gtggtggtac gtaggcgcta ccggtcgcca ccatggtgag
601 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
661 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
721 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
781 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
841 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
901 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
961 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
1021 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
1081 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
1141 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
1201 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
1261 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt ccggactcag
1321 atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcagcg ctgccaacct
1381 gccggccatg gagaccccgt cccagcggcg cgccacccgc agcggggcgc aggccagctc
1441 cactccgctg tcgcccaccc gcatcacccg gctgcaggag aaggaggacc tgcaggagct
1501 caatgatcgc ttggcggtct acatcgaccg tgtgcgctcg ctggaaacgg agaacgcagg
1561 gctgcgcctt cgcatcaccg agtctgaaga ggtggtcagc cgcgaggtgt ccggcatcaa
1621 ggccgcctac gaggccgagc tcggggatgc ccgcaagacc cttgactcag tagccaagga
1681 gcgcgcccgc ctgcagctgg agctgagcaa agtgcgtgag gagtttaagg agctgaaagc
1741 gcgcaatacc aagaaggagg gtgacctgat agctgctcag gctcggctga aggacctgga
1801 ggctctgctg aactccaagg aggccgcact gagcactgct ctcagtgaga agcgcacgct
1861 ggagggcgag ctgcatgatc tgcggggcca ggtggccaag cttgaggcag ccctaggtga
1921 ggccaagaag caacttcagg atgagatgct gcggcgggtg gatgctgaga acaggctgca
1981 gaccatgaag gaggaactgg acttccagaa gaacatctac agtgaggagc tgcgtgagac
2041 caagcgccgt catgagaccc gactggtgga gattgacaat gggaagcagc gtgagtttga
2101 gagccggctg gcggatgcgc tgcaggaact gcgggcccag catgaggacc aggtggagca
2161 gtataagaag gagctggaga agacttattc tgccaagctg gacaatgcca ggcagtctgc
2221 tgagaggaac agcaacctgg tgggggctgc ccacgaggag ctgcagcagt cgcgcatccg
2281 catcgacagc ctctctgccc agctcagcca gctccagaag cagctggcag ccaaggaggc
2341 gaagcttcga gacctggagg actcactggc ccgtgagcgg gacaccagcc ggcggctgct
2401 ggcggaaaag gagcgggaga tggccgagat gcgggcaagg atgcagcagc agctggacga
2461 gtaccaggag cttctggaca tcaagctggc cctggacatg gagatccacg cctaccgcaa
2521 gctcttggag ggcgaggagg agaggctacg cctgtccccc agccctacct cgcagcgcag
2581 ccgtggccgt gcttcctctc actcatccca gacacagggt gggggcagcg tcaccaaaaa
2641 gcgcaaactg gagtccactg agagccgcag cagcttctca cagcacgcac gcactagcgg
2701 gcgcgtggcc gtggaggagg tggatgagga gggcaagttt gtccggctgc gcaacaagtc
2761 caatgaggac cagtccatgg gcaattggca gatcaagcgc cagaatggag atgatccctt
2821 gctgacttac cggttcccac caaagttcac cctgaaggct gggcaggtgg tgacgatctg
2881 ggctgcagga gctggggcca cccacagccc ccctaccgac ctggtgtgga aggcacagaa
2941 cacctggggc tgcgggaaca gcctgcgtac ggctctcatc aactccactg gggaagaagt
3001 ggccatgcgc aagctggtgc gctcagtgac tgtggttgag gacgacgagg atgaggatgg
3061 agatgacctg ctccatcacc accacggctc ccactgcagc agctcggggg accccgctga
3121 gtacaacctg cgctcgcgca ccgtgctgtg cgggacctgc gggcagcctg ccgacaaggc
3181 atctgccagc ggctcaggag cccaggtggg cggacccatc tcctctggct cttctgcctc
3241 cagtgtcacg gtcactcgca gctaccgcag tgtggggggc agtgggggtg gcagcttcgg
3301 ggacaatctg gtcacccgct cctacctcct gggcaactcc agcccccgaa cccagagccc
3361 ccagaactgc agcatcatgt aatctgggac ctgccaggca ggggtggggg tggaggcttc
3421 ctgcgtcctc ctcacctcat gcccaccccc tgccctgcac gtcatgggag ggggcttgaa
3481 gccaaagaaa aataaccctt tggttttttt cttctgtatt tttttttcta agagaagtta
3541 ttttctacag tggttttata ctraaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
3601 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa agcgtctaga aaccctttag atctgctgca
3661 gcgcgccagc acagtggtcg accctgtgga atgtgtgtca gttagggtgt ggaaagtccc
3721 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
3781 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
3841 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
3901 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
3961 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
4021 aaaagcttac catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
4081 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
4141 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
4201 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
4261 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
4321 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
4381 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
4441 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
4501 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
4561 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
4621 ccggtgcctg acgcccgccc cacgacccgc agcgcccgac cgaaaggagc gcacgacccc
4681 atgcatcgat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
4741 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata
4801 cataactgag aatagagaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg
4861 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
4921 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
4981 ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc
5041 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca
5101 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
5161 cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt
5221 atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt
5281 ctcctctgag tgattgacta cccgtcagcg ggggtctttc acatgcagca tgtatcaaaa
5341 ttaatttggt tttttttctt aagtatttac attaaatggc catagttgca ttaatgaatc
5401 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
5461 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
5521 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
5581 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
5641 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
5701 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
5761 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
5821 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
5881 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
5941 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
6001 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
6061 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
6121 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
6181 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
6241 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
6301 atcttcacct agatcctttt gcggccgcaa atcaatctaa agtatatatg agtaaacttg
6361 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
6421 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
6481 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
6541 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
6601 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
6661 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
6721 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
6781 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
6841 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
6901 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
6961 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
7021 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
7081 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
7141 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
7201 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
7261 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
7321 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gcagcctgaa tatgggccaa
7381 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag
7441 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
7501 gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt
7561 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
7621 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
7681 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
7741 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
7801 tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga
7861 gacccctgcc cagggaccac cgacccacca ccgggaggca agctggccag caacttatct
7921 gtgtctgtcc gattgtctag tgtctatgac tgattttatg cgcctgcgtc ggtactagtt
7981 agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttct gaacacccgg
8041 ccgcaaccct gggagacgtc ccagggactt tgggggccgt ttttgtggcc cgacctgagg
8101 aagggagtcg atgtggaatc cgaccccgtc aggatatgtg gttctggtag gagacgagaa
8161 cctaaaacag
//
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