| 质粒名称: | pLOVE-GFP |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 15949 |
| 存储实验室: | Miguel Ramalho-Santos |
|
载体骨架基本信息 |
|
| 载体名称: | pLOVE |
| 空载体大小: | 8263 bp |
| 完整质粒大小: | 7372 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | Stbl3 |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Green Fluorescent Protein/EGFP |
| 物种来源: | Aequorea victoria |
| 插入大小: | 600 bp |
| 基因突变: | The recombination reaction required to clone "Gene X" into the pLOVE vector backbone removes 1456 bp (the CmR-ccdB region). Therefore, the size of pLOVE-GFP is ~7407 bp. |
| 标签/融合蛋白: | V5 (C terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’测序引物 | n/a |
| 3’测序引物 | V5 |
|
文献引用方法 |
|
| 材料和方法部分: | pLOVE-GFP was a gift from Miguel Ramalho-Santos (Addgene plasmid # 15949 ; http://n2t.net/addgene:15949 ; RRID:Addgene_15949) |
| 参考文献部分: | Generation of Induced Pluripotent Stem Cells in the Absence of Drug Selection. Blelloch R, Venere M, Yen J, Ramalho-Santos M. Cell Stem Cell. 2007 Sep 13. ():. 10.1016/j.stem.2007.08.008 PubMed 18371358 |
买家导航
pLOVE-GFP载体质粒基本信息
pLOVE-GFP质粒图谱载体图谱和pLOVE-GFP载体序列质粒序列多克隆位点信息
pLOVE-GFP质粒载体简介
pLOVE-GFP质粒序列载体序列
LOCUS pLOVE-GFP 7372 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pLOVE-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7372)
AUTHORS Blelloch R, Venere M, Yen J, Ramalho-Santos M
TITLE Generation of Induced Pluripotent Stem Cells in the Absence of Drug
Selection.
JOURNAL Cell Stem Cell. 2007 Sep 13. ():.
PUBMED 18371358
REFERENCE 2 (bases 1 to 7372)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7372
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..304
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 305..508
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
protein_bind 510..528
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 531..549
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 590..614
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
regulatory 636..645
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 642..1358
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 642..1358
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(687..708)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(948..967)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1295..1316
/label=EGFP-C
/note="EGFP, forward primer"
protein_bind complement(1390..1414)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 1467..1508
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
protein_bind 1546..1579
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 1635..2223
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(1688..1708)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2106..2117)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(2226..2242)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2227..2243)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 2752..2932
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(2994..3582)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3074..3093)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3753..4613)
/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 4376..4395
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4614..4718)
/gene="bla"
/label=AmpR promoter
primer_bind complement(4793..4812)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 4984..5363
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 5365..5563
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 5581..5761
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 5808..5933
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 6430..6663
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 6848..6892
/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 7190..7307
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
ORIGIN
1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
481 acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc tccctatcag
541 tgatagagat cgtcgactag tccagtgtgg tggaattctg cagatatcaa caagtttgta
601 caaaaaagca ggctttaaag gaaccgctac cggtcgccac catggtgagc aagggcgagg
661 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
721 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
781 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
841 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
901 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
961 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
1021 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
1081 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
1141 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
1201 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
1261 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
1321 ccgccgggat cactctcggc atggacgagc tgtacaagtc cggccggact cagatctcga
1381 gatatctaga cccagctttc ttgtacaaag tggttgatat ccagcacagt ggcggccgct
1441 cgagtctaga gggcccgcgg ttcgaaggta agcctatccc taaccctctc ctcggtctcg
1501 attctacgcg taccggttag taatgacaat tcgtcgaggg acctaataac ttcgtatagc
1561 atacattata cgaagttata catgtttaag ggttccggtt ccactaggta caattcgata
1621 tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
1681 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
1741 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
1801 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
1861 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
1921 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
1981 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
2041 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
2101 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
2161 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
2221 cgcatcgata ccgtcgacct cgatcgagac ctagaaaaac atggagcaat cacaagtagc
2281 aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg
2341 ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat
2401 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
2461 caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga ttggcagaac
2521 tacacaccag ggccagggat cagatatcca ctgacctttg gatggtgcta caagctagta
2581 ccagttgagc aagagaaggt agaagaagcc aatgaaggag agaacacccg cttgttacac
2641 cctgtgagcc tgcatgggat ggatgacccg gagagagaag tattagagtg gaggtttgac
2701 agccgcctag catttcatca catggcccga gagctgcatc cggactgtac tgggtctctc
2761 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
2821 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
2881 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagcatgtga
2941 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
3001 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
3061 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
3121 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
3181 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
3241 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
3301 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
3361 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
3421 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
3481 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
3541 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
3601 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
3661 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
3721 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
3781 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
3841 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
3901 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
3961 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
4021 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
4081 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
4141 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
4201 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
4261 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
4321 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
4381 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
4441 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
4501 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
4561 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
4621 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
4681 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
4741 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
4801 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
4861 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
4921 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
4981 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
5041 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
5101 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
5161 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
5221 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
5281 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
5341 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
5401 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
5461 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
5521 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagcgcgttt tgcctgtact
5581 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
5641 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
5701 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
5761 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
5821 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
5881 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
5941 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
6001 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
6061 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
6121 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
6181 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
6241 aaaacaaaag taagaccacc gcacagcaag cggccggccg ctgatcttca gacctggagg
6301 aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga
6361 accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc
6421 agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc
6481 agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca
6541 gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg
6601 catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct
6661 cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc
6721 tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga
6781 cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca
6841 gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg
6901 gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt
6961 ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata
7021 ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg
7081 aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc
7141 ggcactgcgt gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa
7201 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
7261 tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca
7321 gggacagcag agatccagtt tggttagtac cgggcccgct ctagcagttc cg
//
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pLOVE-GFP质粒载体应用举例
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