| 质粒名称: | LeGO-iG2 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 27341 |
| 存储实验室: | Boris Fehse |
|
载体骨架基本信息 |
|
| 载体名称: | LeGO |
| 空载体大小: | 7829bp |
| 完整质粒大小: | 7829bp |
| 载体类型: | 哺乳细胞表达、Lentiviral |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | SFFV promoter, IRES-eGFP |
| 5’测序引物 | GAGCTCACAACCCCTCACTC |
|
文献引用方法 |
|
| 材料和方法部分: | LeGO-iG2 was a gift from Boris Fehse (Addgene plasmid # 27341 ; http://n2t.net/addgene:27341 ; RRID:Addgene_27341) |
| 参考文献部分: | A multicolor panel of novel lentiviral "gene ontology" (LeGO) vectors for functional gene analysis. Weber K, Bartsch U, Stocking C, Fehse B. Mol Ther. 2008 Apr . 16(4):698-706. 10.1038/mt.2008.6 PubMed 18362927 |
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 LeGO-iG2 7829 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS LeGO-iG2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7829)
AUTHORS Weber K, Bartsch U, Stocking C, Fehse B
TITLE A multicolor panel of novel lentiviral ""gene ontology"" (LeGO)
vectors for functional gene analysis.
JOURNAL Mol Ther. 2008 Apr . 16(4):698-706.
PUBMED 18362927
REFERENCE 2 (bases 1 to 7829)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7829
/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 1686..1919
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2104..2148
/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 2446..2563
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
primer_bind 2622..2638
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2680..2713
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
promoter 2784..3190
/label=SFFV promoter
/note="spleen focus-forming virus long terminal repeat
(LTR) promoter"
misc_feature 3276..3862
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(3443..3460)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 3670..3689
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 3863..4582
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 3863..4579
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(3908..3929)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(4169..4188)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4516..4537
/label=EGFP-C
/note="EGFP, forward primer"
protein_bind 4629..4662
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 4718..5306
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4771..4791)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5189..5200)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(5309..5325)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
LTR 5835..6015
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(6077..6665)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6157..6176)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6836..7696)
/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 7459..7478
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7697..7801)
/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 gccgcgctga tcttcagacc tggaggagga
1561 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
1621 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
1681 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
1741 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
1801 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
1861 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
1921 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
1981 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
2041 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
2101 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
2161 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
2221 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
2281 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
2341 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca
2401 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg
2461 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
2521 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
2581 cagcagagat ccagtttggt tagtaccggg cccgctctag tcgaggtcga cggtatcgat
2641 aagctcgctt cacgagattc cagcaggtcg agggacctaa taacttcgta tagcatacat
2701 tatacgaagt tatattaagg gttccaagct taagcggccg gccgctgaaa gaccccacct
2761 gtaggtttgg caagctagct gcagtaacgc cattttgcaa ggcatggaaa aataccaaac
2821 caagaataga gaagttcaga tcaagggcgg gtacatgaaa atagctaacg ttgggccaaa
2881 caggatatct gcggtgagca gtttcggccc cggcccgggg ccaagaacag atggtcaccg
2941 cagtttcggc cccggcccga ggccaagaac agatggtccc cagatatggc ccaaccctca
3001 gcagtttctt aagacccatc agatgtttcc aggctccccc aaggacctga aatgaccctg
3061 cgccttattt gaattaacca atcagcctgc ttctcgcttc tgttcgcgcg cttctgcttc
3121 ccgagctcta taaaagagct cacaacccct cactcggcgc gccagtcctc cgattgactg
3181 agtcgcccgg atcccagtgt ggtggtacgg gaattcctgc aggcctcgac gagggccggc
3241 gcgccgcggc cgctacgtaa attccgcccc ccccccccct ctccctcccc cccccctaac
3301 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
3361 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
3421 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
3481 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
3541 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
3601 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
3661 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
3721 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
3781 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
3841 cgatgataat atggccacaa ccatggtgag caagggcgag gagctgttca ccggggtggt
3901 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
3961 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
4021 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
4081 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
4141 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
4201 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
4261 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
4321 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
4381 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
4441 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
4501 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
4561 catggacgag ctgtacaagt aaagcggccg gccgccagca cagtggtcga aattcgtcga
4621 gggacctaat aacttcgtat agcatacatt atacgaagtt atacatgttt aagggttccg
4681 gttccactag gtacaattcg atatcaagct tatcgataat caacctctgg attacaaaat
4741 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
4801 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
4861 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
4921 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
4981 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
5041 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
5101 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
5161 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
5221 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
5281 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgatcga gacctagaaa
5341 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag
5401 aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa
5461 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
5521 ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag
5581 gctacttccc tgattggcag aactacacac cagggccagg gatcagatat ccactgacct
5641 ttggatggtg ctacaagcta gtaccagttg agcaagagaa ggtagaagaa gccaatgaag
5701 gagagaacac ccgcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag
5761 aagtattaga gtggaggttt gacagccgcc tagcatttca tcacatggcc cgagagctgc
5821 atccggactg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
5881 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
5941 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
6001 ggaaaatctc tagcagcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
6061 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
6121 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
6181 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
6241 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
6301 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
6361 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
6421 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
6481 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
6541 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
6601 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
6661 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
6721 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
6781 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
6841 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
6901 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
6961 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
7021 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
7081 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
7141 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
7201 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
7261 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
7321 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
7381 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
7441 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
7501 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
7561 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
7621 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
7681 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
7741 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
7801 gcacatttcc ccgaaaagtg ccacctgac
//
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