| 質粒名稱: | pBABE-puro mCherry-EGFP-LC3B |
|---|---|
| 出品公司: | 愛迪基因 |
| 目錄編號: | 22418 |
| 存儲實驗室: | Jayanta Debnath |
|
載體骨架基本信息 |
|
| 載體名稱: | pBABE-puro |
| 載體出品公司: | -- |
| 載體抗性: | 氨芐青霉素 |
| 空載體大小: | 5169 bp |
| 完整質粒大小: | -- |
| 載體修飾: | -- |
| 載體類型: | 哺乳動物表達，逆轉錄病毒 |
| 篩選標記: | 嘌呤霉素 |
| 高拷貝/低拷貝: | 高拷貝 |
| 生長條件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名稱: | microtubule-associated protein 1 light chain 3 beta,MAP1LC3B,LC3B,LC3 |
| 物種來源: | H. sapiens (human) |
| 插入大小: | 1900 bp |
| 基因突變: | -- |
| GenBank編號 | BC045759 |
| 啟動子 | -- |
| 標簽/融合蛋白: |
mCherry（插入件上的N端子）
eGFP（插入件上的N端子）
|
| 克隆方法: | 限制性內切酶 |
| 5' 克隆位點: | BamHI（克隆過程中銷毀） |
| 3' 克隆位點: | SalI (未破壞) |
| 5' 測序引物 | -- |
| 3' 測序引物 | -- |
|
文獻引用方法 |
|
| 材料和方法部分: | pBABE-puro mCherry-EGFP-LC3B was a gift from Jayanta Debnath (Addgene plasmid # 22418 ; //n2t.net/addgene:22418 ; RRID:Addgene_22418) |
| 參考文獻部分: | PLIC proteins or ubiquilins regulate autophagy-dependent cell survival during nutrient starvation. N'Diaye EN, Kajihara KK, Hsieh I, Morisaki H, Debnath J, Brown EJ. EMBO Rep. 2009 Feb;10(2):173-9. doi: 10.1038/embor.2008.238. Epub 2009 Jan 16. 10.1038/embor.2008.238 PubMed 19148225 |
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pBABE-puro mCherry-EGFP-LC3B載體質粒基本信息
pBABE-puro mCherry-EGFP-LC3B質粒圖譜載體圖譜和pBABE-puro mCherry-EGFP-LC3B載體序列質粒序列多克隆位點信息
pBABE-puro mCherry-EGFP-LC3B質粒載體簡介
pBABE-puro mCherry-EGFP-LC3B質粒序列載體序列
LOCUS pBABE-puro mCherry-EGFP-LC3B 6952 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION retroviral expression of mCherry, EGFP-tagged LC3.
ACCESSION .
VERSION .
KEYWORDS pBABE-puro mCherry-EGFP-LC3B
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6952)
AUTHORS N'Diaye EN, Kajihara KK, Hsieh I, Morisaki H, Debnath J, Brown EJ
TITLE PLIC proteins or ubiquilins regulate autophagy-dependent cell
survival during nutrient starvation.
JOURNAL EMBO Rep. 2009 Feb;10(2):173-9. doi: 10.1038/embor.2008.238. Epub
2009 Jan 16.
PUBMED 19148225
REFERENCE 2 (bases 1 to 6952)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
//www.snapgene.com
FEATURES Location/Qualifiers
source 1..6952
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 1..470
/note="long terminal repeat from Moloney murine leukemia
virus"
misc_feature 535..734
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 935..1351
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1281..1303
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1319..1335
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
regulatory 1362..1371
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1368..2075
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(1513..1531)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 1781..1800
/label=mCherry-F
/note="mCherry, forward primer"
protein_bind 2103..2127
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
regulatory 2136..2145
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2142..2858
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(2187..2208)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2448..2467)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2795..2816
/label=EGFP-C
/note="EGFP, forward primer"
CDS 2886..3260
/codon_start=1
/gene="MAP1LC3B"
/product="human LC3 protein"
/label=LC3B
/note="Atg8 family member found on autophagosomes"
/translation="MPSEKTFKQRRTFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLP
VLDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESEKD
EDGFLYMVYASQETFGMKLSV"
primer_bind complement(3275..3295)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3280..3609
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3460..3595
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3522..3541
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3619..4218
/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(3619..3638)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 4115..4135
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 4439..4908
/note="long terminal repeat from Moloney murine leukemia
virus"
primer_bind complement(5038..5055)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5209..5797)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5289..5308)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5957..6817)
/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 6580..6599
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6818..6922)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
61 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
121 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
181 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
241 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
301 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
361 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
421 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct
481 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggcaagctg
541 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct
601 gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
661 gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg gccgtttttg
721 tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat atgtggttct
781 ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg
841 gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt gttgtctctg
901 tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag
961 tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt
1021 caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg
1081 gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc
1141 acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc
1201 ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc
1261 tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct
1321 ttatccagcc ctcactcctt ctctaggcgc cggccggatc tgccaccatg gtgagcaagg
1381 gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg
1441 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
1501 gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
1561 tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc
1621 ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
1681 aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct
1741 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga
1801 ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg
1861 agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca
1921 cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg
1981 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc
2041 gccactccac cggcggcatg gacgagctgt acaagtccgg actcagatcc gctggcccag
2101 gaacaagttt gtacaaaaaa gcaggctttc cggtcgccac catggtgagc aagggcgagg
2161 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
2221 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
2281 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
2341 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
2401 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
2461 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
2521 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
2581 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
2641 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
2701 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
2761 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
2821 ccgccgggat cactctcggc atggacgagc tgtacaagtc cggactcaga tccggtaccg
2881 aattcatgcc gtcggagaag accttcaagc agcgccgcac cttcgaacaa agagtagaag
2941 atgtccgact tattcgagag cagcatccaa ccaaaatccc ggtgataata gaacgataca
3001 agggtgagaa gcagcttcct gttctggata aaacaaagtt ccttgtacct gaccatgtca
3061 acatgagtga gctcatcaag ataattagaa ggcgcttaca gctcaatgct aatcaggcct
3121 tcttcctgtt ggtgaacgga cacagcatgg tcagcgtctc cacaccaatc tcagaggtgt
3181 atgagagtga gaaagatgaa gatggattcc tgtacatggt ctatgcctcc caggagacgt
3241 tcgggatgaa attgtcagtg taagtcgacc ctgtggaatg tgtgtcagtt agggtgtgga
3301 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
3361 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
3421 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
3481 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
3541 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
3601 ttttgcaaaa agcttaccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
3661 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
3721 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
3781 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
3841 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
3901 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
3961 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
4021 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
4081 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
4141 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
4201 cgcaagcccg gtgcctgacg cccgccccac gacccgcagc gcccgaccga aaggagcgca
4261 cgaccccatg catcgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg
4321 aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg
4381 aaaaatacat aactgagaat agagaagttc agatcaaggt caggaacaga tggaacagct
4441 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
4501 acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
4561 ggctcagggc caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag
4621 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
4681 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
4741 aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta
4801 cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg
4861 ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcaca tgcagcatgt
4921 atcaaaatta atttggtttt ttttcttaag tatttacatt aaatggccat agttgcatta
4981 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
5041 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
5101 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
5161 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
5221 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
5281 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
5341 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
5401 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
5461 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
5521 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
5581 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
5641 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
5701 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
5761 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
5821 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
5881 aaaaaggatc ttcacctaga tccttttgcg gccgcaaatc aatctaaagt atatatgagt
5941 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
6001 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
6061 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
6121 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
6181 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
6241 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
6301 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
6361 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
6421 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
6481 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
6541 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
6601 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
6661 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
6721 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
6781 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
6841 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
6901 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgca gc
//