| 质粒名称: | LeGO-EBFP2 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 85213 |
| 存储实验室: | Boris Fehse |
|
载体骨架基本信息 |
|
| 载体名称: | LeGO vector |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6417 bp |
| 完整质粒大小: | 7137 bp |
| 载体类型: | 哺乳动物表达，慢病毒，Cre/Lox |
| 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: | EBFP2 |
| 物种来源: | Synthetic |
| 插入大小: | 720 bp |
| 启动子 | SFFV |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamHI, NcoI (未破坏) |
| 3' 克隆位点: | EcoRI (未破坏) |
| 5' 测序引物 | GAGCTCACAACCCCTCACTC |
|
文献引用方法 |
|
| 材料和方法部分: | LeGO-EBFP2 was a gift from Boris Fehse (Addgene plasmid # 85213 ; http://n2t.net/addgene:85213 ; RRID:Addgene_85213) |
| 参考文献部分: | Optical Barcoding for Single-Clone Tracking to Study Tumor Heterogeneity. Mohme M, Maire CL, Riecken K, Zapf S, Aranyossy T, Westphal M, Lamszus K, Fehse B. Mol Ther. 2017 Jan 18. pii: S1525-0016(16)45496-1. doi: 10.1016/j.ymthe.2016.12.014. 10.1016/j.ymthe.2016.12.014 PubMed 28109958 |
买家导航
LeGOEBFP2载体基本信息
LeGOEBFP2载体图谱质粒图谱和多克隆位点信息
LeGOEBFP2载体简介
LeGOEBFP2载体序列
LOCUS LeGO-EBFP2 7135 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Lentiviral vector expressing EBFP2 fluorescent protein driven by
SFFV promoter.
ACCESSION .
VERSION .
KEYWORDS LeGO-EBFP2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7135)
AUTHORS Mohme M, Maire CL, Riecken K, Zapf S, Aranyossy T, Westphal M,
Lamszus K, Fehse B
TITLE Optical Barcoding for Single-Clone Tracking to Study Tumor
Heterogeneity.
JOURNAL Mol Ther. 2017 Jan 18. pii: S1525-0016(16)45496-1. doi:
10.1016/j.ymthe.2016.12.014.
PUBMED 28109958
REFERENCE 2 (bases 1 to 7135)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7135
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(47..66)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 238..617
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 619..817
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 772..792
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 835..1015
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1062..1187
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1684..1917
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2102..2146
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 2444..2561
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
primer_bind 2620..2636
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2678..2711
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
promoter 2778..3184
/label=SFFV promoter
/note="spleen focus-forming virus long terminal repeat
(LTR) promoter"
regulatory 3190..3199
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3196..3915
/codon_start=1
/product="enhanced blue variant of GFP (Ai et al., 2007)"
/label=EBFP2
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
EFRTAAGITLGMDELYK"
primer_bind complement(3241..3262)
/label=EGFP-N
/note="EGFP, reverse primer"
protein_bind 3935..3968
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 4024..4612
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4077..4097)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4495..4506)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(4615..4631)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
LTR 5141..5321
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(5383..5971)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5463..5482)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6142..7002)
/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 6765..6784
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7003..7107)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
1501 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga
1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
2581 gcagagatcc agtttggtta gtaccgggcc cgctctagtc gaggtcgacg gtatcgataa
2641 gctcgcttca cgagattcca gcaggtcgag ggacctaata acttcgtata gcatacatta
2701 tacgaagtta tattaagggt tccaagctta agcggccgct gaaagacccc acctgtaggt
2761 ttggcaagct agctgcagta acgccatttt gcaaggcatg gaaaaatacc aaaccaagaa
2821 tagagaagtt cagatcaagg gcgggtacat gaaaatagct aacgttgggc caaacaggat
2881 atctgcggtg agcagtttcg gccccggccc ggggccaaga acagatggtc accgcagttt
2941 cggccccggc ccgaggccaa gaacagatgg tccccagata tggcccaacc ctcagcagtt
3001 tcttaagacc catcagatgt ttccaggctc ccccaaggac ctgaaatgac cctgcgcctt
3061 atttgaatta accaatcagc ctgcttctcg cttctgttcg cgcgcttctg cttcccgagc
3121 tctataaaag agctcacaac ccctcactcg gcgcgccagt cctccgattg actgagtcgc
3181 ccggatcccg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
3241 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgagggg cgagggcgag
3301 ggcgatgcca ccaacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
3361 gtgccctggc ccaccctcgt gaccaccctg agccacggcg tgcagtgctt cgcccgctac
3421 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
3481 gagcgcacca tcttcttcaa ggacgacggc acctacaaga cccgcgccga ggtgaagttc
3541 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggcg tcgacttcaa ggaggacggc
3601 aacatcctgg ggcacaagct ggagtacaac ttcaacagcc acaacatcta tatcatggcc
3661 gtcaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacgt ggaggacggc
3721 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
3781 ctgcccgaca gccactacct gagcacccag tccgtgctga gcaaagaccc caacgagaag
3841 cgcgatcaca tggtcctgct ggagttccgc accgccgccg ggatcactct cggcatggac
3901 gagctgtaca agtaagaatt cgtcgaggga cctaataact tcgtatagca tacattatac
3961 gaagttatac atgtttaagg gttccggttc cactaggtac aattcgatat caagcttatc
4021 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
4081 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
4141 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
4201 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
4261 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
4321 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
4381 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
4441 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
4501 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
4561 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac
4621 cgtcgacctc gatcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc
4681 taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt
4741 cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
4801 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatatcct
4861 tgatctgtgg atctaccaca cacaaggcta cttccctgat tggcagaact acacaccagg
4921 gccagggatc agatatccac tgacctttgg atggtgctac aagctagtac cagttgagca
4981 agagaaggta gaagaagcca atgaaggaga gaacacccgc ttgttacacc ctgtgagcct
5041 gcatgggatg gatgacccgg agagagaagt attagagtgg aggtttgaca gccgcctagc
5101 atttcatcac atggcccgag agctgcatcc ggactgtact gggtctctct ggttagacca
5161 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
5221 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
5281 atccctcaga cccttttagt cagtgtggaa aatctctagc agcatgtgag caaaaggcca
5341 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
5401 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
5461 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
5521 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
5581 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
5641 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
5701 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
5761 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
5821 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
5881 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
5941 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
6001 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
6061 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
6121 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
6181 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
6241 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
6301 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
6361 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
6421 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
6481 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
6541 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
6601 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
6661 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
6721 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
6781 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
6841 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
6901 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
6961 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
7021 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
7081 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgac
//
