| 质粒名称: | pLOVE-Klf4 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 15950 |
| 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: | 8055 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: | Kruppel-like factor 4/Klf4, GKLF, EZF |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 2156 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-Klf4 is ~8963 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-Klf4 was a gift from Miguel Ramalho-Santos (Addgene plasmid # 15950 ; http://n2t.net/addgene:15950 ; RRID:Addgene_15950) |
| 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 |
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 pLOVE-Klf4 8055 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pLOVE-Klf4
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8055)
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 8055)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8055
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 382..580
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 598..778
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 825..950
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1447..1680
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1865..1909
/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 2207..2324
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
enhancer 2390..2693
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2694..2897
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
protein_bind 2899..2917
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2920..2938
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2979..3003
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(4462..4486)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 4539..4580
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
protein_bind 4618..4651
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 4707..5295
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4760..4780)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5178..5189)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(5298..5314)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5299..5315)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 5824..6004
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(6066..6654)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6146..6165)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6825..7685)
/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 7448..7467
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7686..7790)
/gene="bla"
/label=AmpR promoter
primer_bind complement(7865..7884)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
1261 acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg atcttcagac ctggaggagg
1321 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
1381 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
1441 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
1501 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
1561 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
1621 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
1681 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
1741 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
1801 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
1861 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
1921 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
1981 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
2041 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat
2101 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc
2161 actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag
2221 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
2281 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
2341 acagcagaga tccagtttgg ttagtaccgg gcccgctcta gcagttccgc gttacataac
2401 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
2461 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
2521 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
2581 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
2641 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
2701 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
2761 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
2821 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
2881 tctatataag cagagctctc cctatcagtg atagagatct ccctatcagt gatagagatc
2941 gtcgactagt ccagtgtggt ggaattctgc agatatcaac aagtttgtac aaaaaagcag
3001 gctccgcggc cgcccccttc accatggctg tcagcgacgc tctgctcccg tccttctcca
3061 cgttcgcgtc cggcccggcg ggaagggaga agacactgcg tccagcaggt gccccgacta
3121 accgttggcg tgaggaactc tctcacatga agcgacttcc cccacttccc ggccgcccct
3181 acgacctggc ggcgacggtg gccacagacc tggagagtgg cggagctggt gcagcttgca
3241 gcagtaacaa cccggccctc ctagcccgga gggagaccga ggagttcaac gacctcctgg
3301 acctagactt tatcctttcc aactcgctaa cccaccagga atcggtggcc gccaccgtga
3361 ccacctcggc gtcagcttca tcctcgtctt ccccagcgag cagcggccct gccagcgcgc
3421 cctccacctg cagcttcagc tatccgatcc gggccggggg tgacccgggc gtggctgcca
3481 gcaacacagg tggagggctc ctctacagcc gagaatctgc gccacctccc acggccccct
3541 tcaacctggc ggacatcaat gacgtgagcc cctcgggcgg cttcgtggct gagctcctgc
3601 ggccggagtt ggacccagta tacattccgc cacagcagcc tcagccgcca ggtggcgggc
3661 tgatgggcaa gtttgtgctg aaggcgtctc tgaccacccc tggcagcgag tacagcagcc
3721 cttcggtcat cagtgttagc aaaggaagcc cagacggcag ccaccccgtg gtagtggcgc
3781 cctacagcgg tggcccgccg cgcatgtgcc ccaagattaa gcaagaggcg gtcccgtcct
3841 gcacggtcag ccggtcccta gaggcccatt tgagcgctgg accccagctc agcaacggcc
3901 accggcccaa cacacacgac ttccccctgg ggcggcagct ccccaccagg actaccccta
3961 cactgagtcc cgaggaactg ctgaacagca gggactgtca ccctggcctg cctcttcccc
4021 caggattcca tccccatccg gggcccaact accctccttt cctgccagac cagatgcagt
4081 cacaagtccc ctctctccat tatcaagagc tcatgccacc gggttcctgc ctgccagagg
4141 agcccaagcc aaagagggga agaaggtcgt ggccccggaa aagaacagcc acccacactt
4201 gtgactatgc aggctgtggc aaaacctata ccaagagttc tcatctcaag gcacacctgc
4261 gaactcacac aggcgagaaa ccttaccact gtgactggga cggctgtggg tggaaattcg
4321 cccgctcgga tgaactgacc aggcactacc gcaaacacac agggcaccgg ccctttcagt
4381 gccagaagtg cgacagggcc ttttccaggt cggaccacct tgccttacac atgaagaggc
4441 actttaaggg tgggcgcgcc gacccagctt tcttgtacaa agtggttgat atccagcaca
4501 gtggcggccg ctcgagtcta gagggcccgc ggttcgaagg taagcctatc cctaaccctc
4561 tcctcggtct cgattctacg cgtaccggtt agtaatgaca attcgtcgag ggacctaata
4621 acttcgtata gcatacatta tacgaagtta tacatgttta agggttccgg ttccactagg
4681 tacaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
4741 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
4801 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
4861 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
4921 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
4981 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
5041 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
5101 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
5161 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
5221 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
5281 gggccgcctc cccgcatcga taccgtcgac ctcgatcgag acctagaaaa acatggagca
5341 atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag
5401 gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag
5461 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
5521 tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct
5581 gattggcaga actacacacc agggccaggg atcagatatc cactgacctt tggatggtgc
5641 tacaagctag taccagttga gcaagagaag gtagaagaag ccaatgaagg agagaacacc
5701 cgcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga agtattagag
5761 tggaggtttg acagccgcct agcatttcat cacatggccc gagagctgca tccggactgt
5821 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
5881 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
5941 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
6001 agcagcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
6061 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
6121 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
6181 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
6241 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
6301 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
6361 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
6421 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
6481 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
6541 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
6601 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
6661 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
6721 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
6781 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
6841 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
6901 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
6961 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
7021 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
7081 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
7141 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
7201 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
7261 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
7321 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
7381 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
7441 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
7501 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
7561 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
7621 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
7681 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
7741 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
7801 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
7861 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
7921 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
7981 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
8041 ccagatatac gcgtt
//
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