| 质粒名称: | pMPRA1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 49349 |
| 存储实验室: | Tarjei Mikkelsen |
|
载体骨架基本信息 |
|
| 载体名称: | pGL4 |
| 载体出品公司: | Promega |
| 载体抗性: | 氯霉素和氨苄青霉素 |
| 空载体大小: | 4238 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 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: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 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: | pMPRA1 was a gift from Tarjei Mikkelsen (Addgene plasmid # 49349 ; http://n2t.net/addgene:49349 ; RRID:Addgene_49349) |
| 参考文献部分: | Massively parallel reporter assays in cultured Mammalian cells. Melnikov A, Zhang X, Rogov P, Wang L, Mikkelsen TS. J Vis Exp. 2014 Aug 17;(90). doi: 10.3791/51719. 10.3791/51719 PubMed 25177895 |
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 pMPRA1 4238 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Basic backbone for transient transfection of Massively Parallel
Reporter Assay (MPRA) libraries.
ACCESSION .
VERSION .
KEYWORDS pMPRA1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4238)
AUTHORS Melnikov A, Zhang X, Rogov P, Wang L, Mikkelsen TS
TITLE Massively parallel reporter assays in cultured Mammalian cells.
JOURNAL J Vis Exp. 2014 Aug 17;(90). doi: 10.3791/51719.
PUBMED 25177895
REFERENCE 2 (bases 1 to 4238)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4238
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 40..164
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 189..219
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 273..932
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(339..358)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
CDS 1274..1579
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
primer_bind 1458..1477
/label=ccdB-fwd
/note="ccdB gene, forward primer"
protein_bind complement(1620..1744)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
polyA_signal 1798..1919
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1809..1828)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 1863..1882
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(2167..2184)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2338..2925)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2418..2437)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3125..3985)
/codon_start=1
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
polyA_signal 4085..4133
/note="synthetic polyadenylation signal"
misc_feature 4147..4238
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
primer_bind 4187..4206
/label=RVprimer3
/note="pGL3 vector, forward primer"
ORIGIN
1 ggcctaactg gccagtgtgg tggaattctg cagatatcaa caagtttgta caaaaaagct
61 gaacgagaaa cgtaaaatga tataaatatc aatatattaa attagatttt gcataaaaaa
121 cagactacat aatactgtaa aacacaacat atccagtcac tatggcggcc gcattaggca
181 ccccaggctt tacactttat gcttccggct cgtataatgt gtggattttg agttaggatc
241 cgtcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca
301 ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc
361 aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga
421 aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc
481 atccggaatt ccgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc
541 cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc
601 acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa
661 acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct
721 gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg
781 ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc
841 aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac
901 agtactgcga tgagtggcag ggcggggcgt aaagatctgg atccggctta ctaaaagcca
961 gataacagta tgcgtatttg cgcgctgatt tttgcggtat aagaatatat actgatatgt
1021 atacccgaag tatgtcaaaa agaggtatgc tatgaagcag cgtattacag tgacagttga
1081 cagcgacagc tatcagttgc tcaaggcata tatgatgtca atatctccgg tctggtaagc
1141 acaaccatgc agaatgaagc ccgtcgtctg cgtgccgaac gctggaaagc ggaaaatcag
1201 gaagggatgg ctgaggtcgc ccggtttatt gaaatgaacg gctcttttgc tgacgagaac
1261 aggggctggt gaaatgcagt ttaaggttta cacctataaa agagagagcc gttatcgtct
1321 gtttgtggat gtacagagtg atattattga cacgcccggg cgacggatgg tgatccccct
1381 ggccagtgca cgtctgctgt cagataaagt ctcccgtgaa ctttacccgg tggtgcatat
1441 cggggatgaa agctggcgca tgatgaccac cgatatggcc agtgtgccgg tctccgttat
1501 cggggaagaa gtggctgatc tcagccaccg cgaaaatgac atcaaaaacg ccattaacct
1561 gatgttctgg ggaatataaa tgtcaggctc ccttatacac agccagtctg caggtcgacc
1621 atagtgactg gatatgttgt gttttacagt attatgtagt ctgtttttta tgcaaaatct
1681 aatttaatat attgatattt atatcatttt acgtttctcg ttcagctttc ttgtacaaag
1741 tggttggcct cggcggccaa gctagtcggg gcggccggcc gcttcgagca gacatgataa
1801 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
1861 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
1921 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
1981 aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatccgtcg accgatgccc
2041 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc
2101 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc
2161 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
2221 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
2281 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
2341 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
2401 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
2461 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
2521 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
2581 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
2641 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
2701 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
2761 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
2821 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
2881 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
2941 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
3001 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
3061 atctaaagta tatatgagta aacttggtct gacagcggcc gcaaatgcta aaccactgca
3121 gtggttacca gtgcttgatc agtgaggcac cgatctcagc gatctgccta tttcgttcgt
3181 ccatagtggc ctgactcccc gtcgtgtaga tcactacgat tcgtgagggc ttaccatcag
3241 gccccagcgc agcaatgatg ccgcgagagc cgcgttcacc ggcccccgat ttgtcagcaa
3301 tgaaccagcc agcagggagg gccgagcgaa gaagtggtcc tgctactttg tccgcctcca
3361 tccagtctat gagctgctgt cgtgatgcta gagtaagaag ttcgccagtg agtagtttcc
3421 gaagagttgt ggccattgct actggcatcg tggtatcacg ctcgtcgttc ggtatggctt
3481 cgttcaactc tggttcccag cggtcaagcc gggtcacatg atcacccata ttatgaagaa
3541 atgcagtcag ctccttaggg cctccgatcg ttgtcagaag taagttggcc gcggtgttgt
3601 cgctcatggt aatggcagca ctacacaatt ctcttaccgt catgccatcc gtaagatgct
3661 tttccgtgac cggcgagtac tcaaccaagt cgttttgtga gtagtgtata cggcgaccaa
3721 gctgctcttg cccggcgtct atacgggaca acaccgcgcc acatagcagt actttgaaag
3781 tgctcatcat cgggaatcgt tcttcggggc ggaaagactc aaggatcttg ccgctattga
3841 gatccagttc gatatagccc actcttgcac ccagttgatc ttcagcatct tttactttca
3901 ccagcgtttc ggggtgtgca aaaacaggca agcaaaatgc cgcaaagaag ggaatgagtg
3961 cgacacgaaa atgttggatg ctcattcctt tttcaatatt attgaagcat ttatcagggt
4021 tactagtacg tctctcaagg ataagtaagt aatattaagg tacgggaggt attggacagg
4081 ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat
4141 agtactaaca tacgctctcc atcaaaacaa aacgaaacaa aacaaactag caaaataggc
4201 tgtccccagt gcaagtgcag gtgccagaac atttctct
//
载体图谱质粒图谱pdf版和相关资料下载
载体质粒应用举例
- 上一篇：pCalN-ddGFP-A
- 下一篇：pCiGSΔΨ
