| 质粒名称: | pSimple-miR302/367 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 98748 |
| 存储实验室: | Sara Howden, James Thomson |
|
载体骨架基本信息 |
|
| 载体名称: | pSimpleII |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | miR302/367 (miRNA cluster) |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1083 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | EF1alpha |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pSimple-miR302/367 was a gift from Sara Howden & James Thomson (Addgene plasmid # 98748 ; http://n2t.net/addgene:98748 ; RRID:Addgene_98748) |
| 参考文献部分: | Simultaneous Reprogramming and Gene Correction of Patient Fibroblasts. Howden SE, Maufort JP, Duffin BM, Elefanty AG, Stanley EG, Thomson JA. Stem Cell Reports. 2015 Dec 8;5(6):1109-18. doi: 10.1016/j.stemcr.2015.10.009. Epub 2015 Nov 12. 10.1016/j.stemcr.2015.10.009 PubMed 26584543 |
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 pSimple-miR302/367 6263 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Episomal reprogramming .
ACCESSION .
VERSION .
KEYWORDS pSimple-miR302/367
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6263)
AUTHORS Howden SE, Maufort JP, Duffin BM, Elefanty AG, Stanley EG, Thomson
JA
TITLE Simultaneous Reprogramming and Gene Correction of Patient
Fibroblasts.
JOURNAL Stem Cell Reports. 2015 Dec 8;5(6):1109-18. doi:
10.1016/j.stemcr.2015.10.009. Epub 2015 Nov 12.
PUBMED 26584543
REFERENCE 2 (bases 1 to 6263)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6263
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 213..1375
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 444..1366
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1323..1343
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
primer_bind complement(2553..2570)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2559..2783
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2948..4061
/label=mini-oriP
/note="internally truncated variant of the Epstein-Barr
virus oriP replication origin (Tanaka et al., 1999)"
primer_bind complement(4106..4122)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4106..4122)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4119..4141)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4130..4146
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(4154..4184)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4199..4220
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4337..4354)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4508..5096)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4588..4607)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5267..6127)
/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 5890..5909
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6128..6232)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg gaggctccgg tgcccgtcag tgggcagagc
241 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct
301 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc
361 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca
421 acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct
481 ttacgggtta tggcccttgc gtgccttgaa ttacttccac ctggctccag tacgtgattc
541 ttgatcccga gctggagcca ggggcgggcc ttgcgcttta ggagcccctt cgcctcgtgc
601 ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg
661 cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc
721 gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caggatctgc acactggtat
781 ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc
841 gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg
901 gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct
961 ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctccagg
1021 gggctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag
1081 gaaaggggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc
1141 gtccaggcac ctcgattagt tctggagctt ttggagtacg tcgtctttag gttgggggga
1201 ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc
1261 ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat
1321 tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgaacacg
1381 tggtcgcggc caattctgca gtcgacggta ccgcgggccc gggatccgga attcccatcg
1441 atggcctcga gaaagcggcc gcagctgcta gcgtccctgc cttttaccct tctggaggag
1501 aacacgaatc tttgggaact agttcaggaa ggtaaaaaaa tttttttctt ctaaagttat
1561 gccattttgt tttctttctc ctcagctcta aatactctga agtccaaaga agttgtatgt
1621 tgggtgggct cccttcaact ttaacatgga agtgctttct gtgactttaa aagtaagtgc
1681 ttccatgttt tagtaggagt gaatccaatt tacttctcca aaatagaaca cgctaacctc
1741 atttgaaggg atcccctttg ctttaacatg ggggtacctg ctgtgtgaaa caaaagtaag
1801 tgcttccatg tttcagtgga ggtgtctcca agccagcaca ccttttgtta caaaattttt
1861 ttgttattgt gttttaaggt tactaagctt gttacaggtt aaaggattct aactttttcc
1921 aagactgggc tccccaccac ttaaacgtgg atgtacttgc tttgaaacta aagaagtaag
1981 tgcttccatg ttttggtgat ggtaagtctt ccttttacat ttttattatt tttttagaaa
2041 ataactttat tgtattgacc gcagctcata tatttaagct ttattttgta tttttacatc
2101 tgttaagggg ccccctctac tttaacatgg aggcacttgc tgtgacatga caaaaataag
2161 tgcttccatg tttgagtgtg gtggttccta cctaatcagc aattgagtta acgcccacac
2221 tgtgtgcagt tcttggctac aggccattac tgttgctaat atgcaactct gttgaatata
2281 aattggaatt gcactttagc aatggtgatg gattgttaag ccaatgacag aatttaaacc
2341 acagacttac tttgatagca ctcttaatgg tataacttct tctcccattt tatgtctctc
2401 tttatgtttt ttcttatgtt tcctttttgt tttcaagaga gagctatctt ttagatctcc
2461 agtatccttt tcctcttcag tagagatttc tatatcttga aataaataaa aaagctctcc
2521 gttaagacat gggtctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
2581 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
2641 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
2701 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
2761 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctcgaccga
2821 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg
2881 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccctggccgg
2941 ggtcccgcag gaaaaggaca agcagcgaaa attcacgccc ccttgggagg tggcggcata
3001 tgcaaaggat agcactccca ctctactact gggtatcata tgctgactgt atatgcatga
3061 ggatagcata tgctacccgg atacagatta ggatagcata tactacccag atatagatta
3121 ggatagcata tgctacccag atatagatta ggatagccta tgctacccag atataaatta
3181 ggatagcata tactacccag atatagatta ggatagcata tgctacccag atatagatta
3241 ggatagccta tgctacccag atatagatta ggatagcata tgctacccag atatagatta
3301 ggatagcata tgctatccag atatttgggt agtatatgct acccagatat aaattaggat
3361 agcatatact accctaatct ctattaggat agcatatgct acccggatac agattaggat
3421 agcatatact acccagatat agattaggat agcatatgct acccagatat agattaggat
3481 agcctatgct acccagatat aaattaggat agcatatact acccagatat agattaggat
3541 agcatatgct acccagatat agattaggat agcctatgct acccagatat agattaggat
3601 agcatatgct atccagatat ttgggtagta tatgctatcc atggcaacat tagcccaccg
3661 tgctctcagc gacctcgtga atatgaggac caacaaccct gtgcttggcg ctcaggcgca
3721 agtgtgtgta atttgtcctc cagatcgcag caatcgcgcc cctatcttgg cccgcccacc
3781 tacttatgca ggtattcccc ggggtgccat tagtggtttt gtgggcaagt ggtttgaccg
3841 cagtggttag cggggttaca atcagccaag ttattacacc cttattttac agtccaaaac
3901 cgcagggcgg cgtgtggggg ctgacgcgat cgctgttcct taggaccctt ttactaaccc
3961 taattcgata gcatatgctt cccgttgggt aacatatgct attgaattag ggttagtctg
4021 gatagtatat actactaccc gggaagcata tgctacccgt ttagggttta ccgtcgacct
4081 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
4141 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
4201 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
4261 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
4321 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
4381 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
4441 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
4501 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
4561 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
4621 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
4681 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
4741 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
4801 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
4861 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
4921 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
4981 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
5041 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
5101 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
5161 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
5221 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
5281 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
5341 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
5401 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
5461 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
5521 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
5581 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
5641 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
5701 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
5761 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
5821 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
5881 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
5941 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
6001 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
6061 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
6121 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
6181 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
6241 cccgaaaagt gccacctgac gtc
//
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