| 质粒名称: | pLLU2G-Oct3 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 21616 |
| 存储实验室: | David Garbers |
|
载体骨架基本信息 |
|
| 载体名称: | pLLU 2G |
| 空载体大小: | 7861 bp |
| 完整质粒大小: | 8318bp |
| 载体类型: | Lentiviral, RNAi |
| 筛选标记: | EGFP |
| 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: | Oct3 shRNA /Oct4, Pou5f1 |
| 插入大小: | 54 bp |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | HpaI(未破坏) |
| 3’克隆位点: | HpaI (未破坏) |
| 5’测序引物 | hUBCpro-F |
| 3’测序引物 | EGFP-N |
|
文献引用方法 |
|
| 材料和方法部分: | pLLU2G-Oct3 was a gift from David Garbers (Addgene plasmid # 21616 ; http://n2t.net/addgene:21616 ; RRID:Addgene_21616) |
| 参考文献部分: | Spermatogonial stem cell self-renewal requires OCT4, a factor downregulated during retinoic acid-induced differentiation. Dann CT, Alvarado AL, Molyneux LA, Denard BS, Garbers DL, Porteus MH. Stem Cells. 2008 Nov . 26(11):2928-37. 10.1634/stemcells.2008-0134 PubMed 18719224 |
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 pLLU2G-Oct3 8318 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pLLU2G-Oct3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8318)
AUTHORS Dann CT, Alvarado AL, Molyneux LA, Denard BS, Garbers DL, Porteus MH
TITLE Spermatogonial stem cell self-renewal requires OCT4, a factor
downregulated during retinoic acid-induced differentiation.
JOURNAL Stem Cells. 2008 Nov . 26(11):2928-37.
PUBMED 18719224
REFERENCE 2 (bases 1 to 8318)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8318
/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"
promoter 2622..2935
/label=U6 promoter
/note="RNA polymerase III promoter for mouse U6 snRNA (Das
et al., 1988)"
primer_bind 2854..2874
/label=mU6-F
/note="Mouse U6 promoter, forward primer"
primer_bind 3006..3022
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3064..3097
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
promoter 3140..4349
/label=UbC promoter
/note="human ubiquitin C promoter"
primer_bind 4186..4205
/label=hUBCpro-F
/note="Human Ubiquitin C (UbC) promoter, forward primer"
regulatory 4373..4382
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 4379..5095
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(4424..4445)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(4685..4704)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 5032..5053
/label=EGFP-C
/note="EGFP, forward primer"
protein_bind 5118..5151
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 5207..5795
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5260..5280)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5678..5689)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(5798..5814)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
LTR 6324..6504
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(6566..7154)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6646..6665)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7325..8185)
/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 7948..7967
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8186..8290)
/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 agatccgacg ccgccatctc
2641 taggcccgcg ccggccccct cgcacagact tgtgggagaa gctcggctac tcccctgccc
2701 cggttaattt gcatataata tttcctagta actatagagg cttaatgtgc gataaaagac
2761 agataatctg ttctttttaa tactagctac attttacatg ataggcttgg atttctataa
2821 gagatacaaa tactaaatta ttattttaaa aaacagcaca aaaggaaact caccctaact
2881 gtaaagtaat tgtgtgtttt gagactataa atatcccttg gagaaaagcc ttgtttgaac
2941 cgtgtgaggt ggaacttcaa gagagttcca cctcacacgg ttctttttta acgcgcggtg
3001 accctcgagg tcgacggtat cgataagctc gcttcacgag attccagcag gtcgagggac
3061 ctaataactt cgtatagcat acattatacg aagttatatt aagggttcca agcttaagcg
3121 gcccgcgggc ccgggatctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc
3181 ctcctcacgg cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct gatccttccg
3241 cccggacgct caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc
3301 agcagaagga cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag
3361 agagcggaac aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt
3421 ggggcggtga acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt
3481 cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga
3541 gtagcgggct gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga
3601 agcgtgtgga gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac
3661 tgggggttgg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc
3721 ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac
3781 ccaaggtctt gaggccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg
3841 gcaccatctg gggaccctga cgtgaagttt gtcactgact ggagaactcg gtttgtcgtc
3901 tgttgcgggg gcggcagtta tgcggtgccg ttgggcagtg cacccgtacc tttgggagcg
3961 cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc
4021 acctgccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag
4081 ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc
4141 gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt
4201 gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa
4261 tgtaatcatt tgggtcaata tgtaattttc agtgttagac tagtaaattg tccgctaaat
4321 tctggccgtt tttggctttt ttgttagacg aagcttgcta gcgctaccgg tcgccaccat
4381 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
4441 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
4501 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
4561 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
4621 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
4681 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
4741 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
4801 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
4861 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
4921 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
4981 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
5041 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtagga
5101 attcgtcgag ggacctaata acttcgtata gcatacatta tacgaagtta tacatgttta
5161 agggttccgg ttccactagg tacaattcga tatcaagctt atcgataatc aacctctgga
5221 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
5281 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
5341 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
5401 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
5461 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
5521 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
5581 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
5641 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
5701 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
5761 gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac ctcgatcgag
5821 acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg
5881 cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt
5941 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
6001 gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc
6061 acacacaagg ctacttccct gattggcaga actacacacc agggccaggg atcagatatc
6121 cactgacctt tggatggtgc tacaagctag taccagttga gcaagagaag gtagaagaag
6181 ccaatgaagg agagaacacc cgcttgttac accctgtgag cctgcatggg atggatgacc
6241 cggagagaga agtattagag tggaggtttg acagccgcct agcatttcat cacatggccc
6301 gagagctgca tccggactgt actgggtctc tctggttaga ccagatctga gcctgggagc
6361 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
6421 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
6481 agtcagtgtg gaaaatctct agcagcatgt gagcaaaagg ccagcaaaag gccaggaacc
6541 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
6601 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
6661 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
6721 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
6781 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
6841 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
6901 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
6961 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
7021 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
7081 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
7141 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
7201 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
7261 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
7321 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
7381 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
7441 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
7501 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
7561 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
7621 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
7681 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
7741 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
7801 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
7861 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
7921 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
7981 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
8041 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
8101 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
8161 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
8221 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
8281 gggttccgcg cacatttccc cgaaaagtgc cacctgac
//
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