| 质粒名称: | pcdna6.2 emgfp mir9 |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 22741 |
| 存储实验室: | Lynn Hudson |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA-6.2 GW/EmGFP miR-neg control |
| 载体出品公司: | Invitrogen |
| 空载体大小: | 5699 bp |
| 完整质粒大小: | 5763 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 壮观霉素，50ug/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: | microRNA 9/mir9 |
| 插入大小: | 951 bp |
| 克隆方法: | 限制性内切酶 |
| 5’测序引物 | 未知 |
| 3’测序引物 | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA6.2-EmGFP-mir9 was a gift from Lynn Hudson (Addgene plasmid # 22741 ; http://n2t.net/addgene:22741 ; RRID:Addgene_22741) |
| 参考文献部分: | Identification of dynamically regulated microRNA and mRNA networks in developing oligodendrocytes. Lau P, Verrier JD, Nielsen JA, Johnson KR, Notterpek L, Hudson LD. J Neurosci. 2008 Nov 5. 28(45):11720-30. 10.1523/JNEUROSCI.1932-08.2008 PubMed 18987208 |
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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 pcDNA6.2-emGFP-mir9 5763 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pcDNA6.2-EmGFP-mir9
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5763)
AUTHORS Lau P, Verrier JD, Nielsen JA, Johnson KR, Notterpek L, Hudson LD
TITLE Identification of dynamically regulated microRNA and mRNA networks
in developing oligodendrocytes.
JOURNAL J Neurosci. 2008 Nov 5. 28(45):11720-30.
PUBMED 18987208
REFERENCE 2 (bases 1 to 5763)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5763
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 4..383
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 384..587
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 538..558
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
promoter 632..650
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 680..704
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 713..1432
/codon_start=1
/product="Emerald GFP"
/label=EmGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(758..779)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1019..1038)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1366..1387
/label=EGFP-C
/note="EGFP, forward primer"
ncRNA 1492..1518
/label=5' miR-155
/note="sequence upstream of the precursor of mouse miR-155
microRNA (Uva et al., 2013)"
/ncRNA_class="miRNA"
ncRNA 1586..1627
/label=3' miR-155
/note="sequence downstream of the precursor of mouse
miR-155 microRNA (Uva et al., 2013)"
/ncRNA_class="miRNA"
protein_bind complement(1656..1680)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
primer_bind complement(1722..1741)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 1766..1814
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
promoter complement(1985..2003)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2008..2024)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2008..2024)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
rep_origin 2092..2520
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2179..2198)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2389..2410
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2529..2549)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2534..2863
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2714..2849
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2776..2795
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 2911..2958
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 2977..3375
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=BSD
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
polyA_signal 3533..3654
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3570..3589)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3624..3643
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3689..3706)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3860..4448)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3940..3959)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4544..5335)
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
1 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
361 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
421 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
481 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
541 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta
601 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagtcccaag
661 ctggctagtt aagctatcaa caagtttgta caaaaaagca ggctttaaaa ccatggtgag
721 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
781 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
841 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
901 caccttcacc tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
961 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
1021 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
1081 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
1141 gtacaactac aacagccaca aggtctatat caccgccgac aagcagaaga acggcatcaa
1201 ggtgaacttc aagacccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
1261 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
1321 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
1381 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aagctaagca
1441 cttcgtggcc gtcgatcgtt taaagggagg tagtgagtcg accagtggat cctggaggct
1501 tgctgaaggc tgtatgctgt ctttggttat ctagctgtat gagttttggc cactgactga
1561 ctcatacaga gataaccaaa gacaggacac aaggcctgtt actagcactc acatggaaca
1621 aatggcccag atctggccgc actcgagata tctagaccca gctttcttgt acaaagtggt
1681 tgatctagag ggcccgcggt tcgctgatgg gggaggctaa ctgaaacacg gaaggagaca
1741 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt
1801 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
1861 accgtgaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
1921 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
1981 atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg
2041 tctcaaaatc tctgatggat ctgcgcagct ggggctctag ggggtatccc cacgcgccct
2101 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
2161 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
2221 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
2281 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
2341 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
2401 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
2461 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
2521 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
2581 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
2641 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
2701 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
2761 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
2821 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
2881 tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc
2941 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagccttt gtctcaagaa
3001 gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac
3061 tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta
3121 tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg
3181 gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc
3241 ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg
3301 aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat
3361 gtgtgggagg gctaagcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt
3421 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
3481 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
3541 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
3601 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
3661 ctgtataccg tcgctcttcc gctgcttcct cgctcactga ctcgctgcgc tcggtcgttc
3721 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
3781 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
3841 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
3901 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
3961 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
4021 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
4081 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
4141 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
4201 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
4261 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
4321 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
4381 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
4441 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgacgcgt
4501 aactcacgtt aagggatttt ggtcatgggt ggctcgacga gggttatttg ccgactacct
4561 tggtgatctc gcctttcacg tagtggacaa attcttccaa ctgatctgcg cgcgaggcca
4621 agcgatcttc ttcttgtcca agataagcct gtctagcttc aagtatgacg ggctgatact
4681 gggccggcag gcgctccatt gcccagtcgg cagcgacatc cttcggcgcg attttgccgg
4741 ttactgcgct gtaccaaatg cgggacaacg taagcactac atttcgctca tcgccagccc
4801 agtcgggcgg cgagttccat agcgttaagg tttcatttag cgcctcaaat agatcctgtt
4861 caggaaccgg atcaaagagt tcctccgccg ctggacctac caaggcaacg ctatgttctc
4921 ttgcttttgt cagcaagata gccagatcaa tgtcgatcgt ggctggctcg aagatacctg
4981 caagaatgtc attgcgctgc cattctccaa attgcagttc gcgcttagct ggataacgcc
5041 acggaatgat gtcgtcgtgc acaacaatgg tgacttctac agcgcggaga atctcgctct
5101 ctccagggga agccgaagtt tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat
5161 caagccttac ggtcaccgta accagcaaat caatatcact gtgtggcttc aggccgccat
5221 ccactgcgga gccgtacaaa tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga
5281 cgccaactac ctctgatagt tgagtcgata cttcggcgat caccgcttcc ctcataatgt
5341 ttaactttgt tttagggcga ctgccctgct gcgtaacatc gttgctgctc cataacatca
5401 aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc ccgaggcata gactgtaccc
5461 caaaaaaaca gtcataacaa gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt
5521 cggtcaaggt tctggaccag ttgcgtgagc gcatacgcta cttgcattac agcttacgaa
5581 ccgaacaggc ttatgtccac tgggttcgtg ccttcatccg tttccacggt gtgcgtcacc
5641 cggcaacctt gggtagcagc gaagtcgagg catttctgtc ctggctggtc tagaattgca
5701 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
5761 cgc
//
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