| 质粒名称: | pcDNA3-tRNA scaffolded SA (tRSA) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 32200 |
| 存储实验室: | Ian Macara |
|
载体骨架基本信息 |
|
| 载体名称: | pDNA3 |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | XL1 Blue |
| 拷贝数: | 高拷贝 |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3-tRNA scaffolded SA (tRSA) was a gift from Ian Macara (Addgene plasmid # 32200 ; http://n2t.net/addgene:32200 ; RRID:Addgene_32200) |
| 参考文献部分: | Efficient detection of RNA-protein interactions using tethered RNAs. Iioka H, Loiselle D, Haystead TA, Macara IG. Nucleic Acids Res. 2011 Apr;39(8):e53. Epub 2011 Feb 7. 10.1093/nar/gkq1316 PubMed 21300640 |
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
载体图谱.png)
载体简介
载体序列
LOCUS pcDNA3-tRNA scaffolded SA (tRSA) 5593 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pcDNA3-tRNA scaffolded SA (tRSA)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5593)
AUTHORS Iioka H, Loiselle D, Haystead TA, Macara IG
TITLE Efficient detection of RNA-protein interactions using tethered RNAs.
JOURNAL Nucleic Acids Res. 2011 Apr;39(8):e53. Epub 2011 Feb 7.
PUBMED 21300640
REFERENCE 2 (bases 1 to 5593)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5593
/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"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(1145..1163)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(1146..1163)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(1183..1200)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1189..1413
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1459..1887
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1546..1565)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1756..1777
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(1896..1916)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 1901..2231
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2082..2217
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2144..2163
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2298..3092
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(2352..2371)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 2962..2981
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3266..3387
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3303..3322)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3357..3376
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3436..3452)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3436..3452)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3449..3471)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3460..3476
/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(3484..3514)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3529..3550
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3667..3684)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3838..4426)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3918..3937)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4597..5457)
/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 5220..5239
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5458..5562)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttgaaaaaaa aaaaagcccg
961 gatagctcag tcggtagagc agcggcctcg accagaatca tgcaagtgcg taagatagtc
1021 gcgggtcgag gccgcgtcca gggttcaagt ccctgttcgg gcgccactgc agaaaaaaaa
1081 aaaagaattc tgcagatatc catcacactg gcggccgctc gagcatgcat ctagagggcc
1141 ctattctata gtgtcaccta aatgctagag ctcgctgatc agcctcgact gtgccttcta
1201 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
1261 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
1321 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
1381 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
1441 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
1501 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
1561 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc
1621 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
1681 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
1741 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
1801 tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc
1861 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
1921 aaagtcccca ggctccccag gcaggcagaa gtatgcaaag catgcatctc aattagtcag
1981 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
2041 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
2101 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
2161 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
2221 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
2281 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
2341 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
2401 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
2461 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
2521 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
2581 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
2641 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
2701 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
2761 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
2821 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
2881 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
2941 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
3001 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
3061 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
3121 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
3181 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
3241 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
3301 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
3361 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
3421 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
3481 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
3541 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
3601 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
3661 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
3721 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
3781 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
3841 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
3901 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
3961 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
4021 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
4081 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
4141 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
4201 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
4261 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
4321 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
4381 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
4441 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
4501 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
4561 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
4621 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
4681 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
4741 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
4801 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
4861 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
4921 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
4981 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
5041 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
5101 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
5161 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
5221 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
5281 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
5341 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
5401 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
5461 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
5521 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
5581 gccacctgac gtc
//
