| 质粒名称: | pCALSL-mir30 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 13786 |
| 存储实验室: | Connie Cepko |
|
载体骨架基本信息 |
|
| 载体名称: | pCAGGS |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5555 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，Cre/Lox |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | mir30 cassette |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 205 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | XhoI（在克隆过程中被破坏） |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | pCAG-F |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pCALSL-mir30 was a gift from Connie Cepko (Addgene plasmid # 13786 ; http://n2t.net/addgene:13786 ; RRID:Addgene_13786) |
| 参考文献部分: | Controlled expression of transgenes introduced by in vivo electroporation. Matsuda T, Cepko CL. Proc Natl Acad Sci U S A. 2007 Jan 5. ():. 10.1073/pnas.0610155104 PubMed 17209010 |
买家导航
pCALSLmir30载体基本信息
pCALSLmir30载体图谱质粒图谱和多克隆位点信息
pCALSLmir30载体简介
pCALSLmir30载体序列
LOCUS pCALSL-mir30 5759 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCALSL-mir30
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5759)
AUTHORS Matsuda T, Cepko CL
TITLE Controlled expression of transgenes introduced by in vivo
electroporation.
JOURNAL Proc Natl Acad Sci U S A. 2007 Jan 5. ():.
PUBMED 17209010
REFERENCE 2 (bases 1 to 5759)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5759
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 4..383
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 386..661
/label=chicken beta-actin promoter
intron 662..1670
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1594..1616
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
polyA_signal 1898..1979
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
polyA_signal 2125..2206
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
polyA_signal 2249..2383
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
protein_bind 2392..2425
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
ncRNA 2463..2556
/label=5' miR-30a
/note="sequence upstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)"
/ncRNA_class="miRNA"
ncRNA 2570..2682
/label=3' miR-30a
/note="sequence downstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)"
/ncRNA_class="miRNA"
primer_bind complement(2783..2802)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_site 2848..2903
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(2902..2921)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(3262..3278)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3262..3278)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3275..3297)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3286..3302
/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(3310..3340)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3355..3376
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3435..3630
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 3481..3616
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3543..3562
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 3636..3770
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3838..3855)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4009..4597)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4089..4108)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4768..5628)
/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 5391..5410
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5629..5733)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
1681 acgtgctggt tgttgtgctg tctcatcatt ttggcaaaga attgattaat tcgagcgaac
1741 gcgtactagt ataacttcgt atagcataca ttatacgagt tattttttga gctccataat
1801 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
1861 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
1921 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
1981 ttctagttgt ggtttgtcca aactcatcaa tgtatccgcg gcataatcag ccataccaca
2041 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
2101 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
2161 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
2221 ttgtccaaac tcatcaatgt atatcgataa cttgtttatt gcagcttata atggttacaa
2281 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
2341 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaaatccg aataacttcg
2401 tatagcatac attatacgaa gttatactag tctcgatcga ctagggataa cagggtaatt
2461 gtttgaatga ggcttcagta ctttacagaa tcgttgcctg cacatcttgg aaacacttgc
2521 tgggattact tcttcaggtt aacccaacag aaggctcgag caaccagaat tcaaggggct
2581 actttaggag caattatctt gtttactaaa actgaatacc ttgctatctc tttgatacat
2641 ttttacaaag ctgaattaaa atggtataaa ttaaatcact tttttgatat catcgatgcg
2701 gccgcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc
2761 tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga
2821 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt
2881 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac
2941 atcagaatga gtatttggtt tagagtttgg caacatatgc catatgctgg ctgccatgaa
3001 caaaggtggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
3061 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
3121 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc
3181 tcctgactac tcccagtcat agctgtccct cttctcttat gaagatccct cgacctgcag
3241 cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
3301 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
3361 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
3421 gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
3481 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
3541 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
3601 ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg
3661 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
3721 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta
3781 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
3841 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
3901 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
3961 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
4021 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
4081 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
4141 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
4201 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
4261 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
4321 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
4381 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
4441 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
4501 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
4561 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
4621 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
4681 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
4741 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
4801 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
4861 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
4921 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
4981 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
5041 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
5101 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
5161 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
5221 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
5281 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
5341 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
5401 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
5461 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
5521 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
5581 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
5641 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
5701 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctg
//
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