| 质粒名称: | pSynSRE-T-Luc |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 60444 |
| 存储实验室: | Timothy Osborne |
|
载体骨架基本信息 |
|
| 载体名称: | pGL2 basic |
| 载体出品公司: | Promega |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5357 bp |
| 完整质粒大小: | 5504 bp |
| 载体修饰: | 包含仓鼠HMG-CoA合成酶启动子序列，该启动子序列含有SREBP反应区（-324/-225），直接与连接到pGL2碱性多聚体中荧光素酶的最小HMG-CoA合成酶TATA盒（-28/+39）融合 |
| 载体类型: | 哺乳动物表达，荧光素酶 |
| 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: | HMG-CoA synthase promoter (-324/-225),3-Hydroxy-3-methylglutaryl-coenzyme A synthase promoter |
| 物种来源: | Hamster |
| 插入大小: | 186 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | HMG-CoA synthase (-324/-225) & TATAA (-28/+39) |
| 标签/融合蛋白: | Luciferase（主干上的C末端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | SstI or SacI(未破坏) |
| 3' 克隆位点: | HindIII用于启动子和TATAA；NheI仅用于启动子区域（未破坏） |
| 5' 测序引物 | GLprimer1 |
| 3' 测序引物 | GLprimer2 |
|
文献引用方法 |
|
| 材料和方法部分: | pSynSRE-T-Luc was a gift from Timothy Osborne (Addgene plasmid # 60444 ; http://n2t.net/addgene:60444 ; RRID:Addgene_60444) |
| 参考文献部分: | Sterol regulation of 3-hydroxy-3-methylglutaryl-coenzyme A synthase gene through a direct interaction between sterol regulatory element binding protein and the trimeric CCAAT-binding factor/nuclear factor Y. Dooley KA, Millinder S, Osborne TF. J Biol Chem. 1998 Jan 16;273(3):1349-56. PubMed 9430668 |
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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 pSynSRE-T-Luc 5757 bp ds-DNA circular SYN 10-DEC-2019
DEFINITION Uses the hamster HMG-CoA synthase promoter sequence (-324/-225) and
minimal HMG-CoA sythnase TATA box (-28/+39) to study SREBP-dependent
transcriptional activation..
ACCESSION .
VERSION .
KEYWORDS pSynSRE-T-Luc
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5757)
AUTHORS Dooley KA, Millinder S, Osborne TF
TITLE Sterol regulation of 3-hydroxy-3-methylglutaryl-coenzyme A synthase
gene through a direct interaction between sterol regulatory element
binding protein and the trimeric CCAAT-binding factor/nuclear factor
Y.
JOURNAL J Biol Chem. 1998 Jan 16;273(3):1349-56.
PUBMED 9430668
REFERENCE 2 (bases 1 to 5757)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Dec 10, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5757
/organism="synthetic DNA construct"
/mol_type="other DNA"
polyA_signal 438..572
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(714..731)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(885..1473)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(965..984)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1644..2504)
/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 2267..2286
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2505..2609)
/gene="bla"
/label=AmpR promoter
rep_origin 2636..3091
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2723..2742)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2933..2954
/label=F1ori-F
/note="F1 origin, forward primer"
polyA_signal 3281..3402
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
CDS 3662..5314
/codon_start=1
/gene="luc"
/product="firefly luciferase"
/label=luciferase
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
primer_bind complement(3728..3746)
/label=LucNrev
/note="Firefly luciferase, reverse primer"
intron 5555..5620
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 5750..13
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
ORIGIN
1 gaagagaaag gtagaagacc ccaaggactt tccttcagaa ttgctaagtt ttttgagtca
61 tgctgtgttt agtaatagaa ctcttgcttg ctttgctatt tacaccacaa aggaaaaagc
121 tgcactgcta tacaagaaaa ttatggaaaa atattctgta acctttataa gtaggcataa
181 cagttataat cataacatac tgttttttct tactccacac aggcatagag tgtctgctat
241 taataactat gctcaaaaat tgtgtacctt tagcttttta atttgtaaag gggttaataa
301 ggaatatttg atgtatagtg ccttgactag agatcataat cagccatacc acatttgtag
361 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
421 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
481 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
541 aactcatcaa tgtatcttat catgtctgga tccgtcgacc gatgcccttg agagccttca
601 acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg
661 tcttctttat catgcaactc gtaggacagg tgccggcagc gctcttccgc ttcctcgctc
721 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
781 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
841 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
901 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
961 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
1021 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
1081 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
1141 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
1201 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
1261 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
1321 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
1381 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
1441 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
1501 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
1561 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
1621 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
1681 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
1741 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
1801 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
1861 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
1921 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
1981 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
2041 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
2101 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
2161 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
2221 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
2281 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
2341 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
2401 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
2461 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
2521 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
2581 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg
2641 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
2701 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
2761 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
2821 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
2881 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
2941 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
3001 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
3061 aattttaaca aaatattaac gcttacaatt tgccattcgc cattcaggct gcgcaactgt
3121 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agcccaagct accatgataa
3181 gtaagtaata ttaaggtacg tggaggtttt acttgcttta aaaaacctcc cacacctccc
3241 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
3301 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
3361 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatggtactg taactgagct
3421 aacataaccc gggaggtacc gagctctggc tctcgccgtc tctcgccgtc ccgccattgg
3481 cctagatgtt ctcgggccac cctacgtcac cgtctcaccc cacttccccg gtcctccgac
3541 tttgcgctag ctcgacaagg tcaactataa agctgctgag ggcgggaggc acagtgcact
3601 gtgctccgtg gattgctcct ttctttgccc ccaagcttgg cattccggta ctgttggtaa
3661 aatggaagac gccaaaaaca taaagaaagg cccggcgcca ttctatcctc tagaggatgg
3721 aaccgctgga gagcaactgc ataaggctat gaagagatac gccctggttc ctggaacaat
3781 tgcttttaca gatgcacata tcgaggtgaa catcacgtac gcggaatact tcgaaatgtc
3841 cgttcggttg gcagaagcta tgaaacgata tgggctgaat acaaatcaca gaatcgtcgt
3901 atgcagtgaa aactctcttc aattctttat gccggtgttg ggcgcgttat ttatcggagt
3961 tgcagttgcg cccgcgaacg acatttataa tgaacgtgaa ttgctcaaca gtatgaacat
4021 ttcgcagcct accgtagtgt ttgtttccaa aaaggggttg caaaaaattt tgaacgtgca
4081 aaaaaaatta ccaataatcc agaaaattat tatcatggat tctaaaacgg attaccaggg
4141 atttcagtcg atgtacacgt tcgtcacatc tcatctacct cccggtttta atgaatacga
4201 ttttgtacca gagtcctttg atcgtgacaa aacaattgca ctgataatga attcctctgg
4261 atctactggg ttacctaagg gtgtggccct tccgcataga actgcctgcg tcagattctc
4321 gcatgccaga gatcctattt ttggcaatca aatcattccg gatactgcga ttttaagtgt
4381 tgttccattc catcacggtt ttggaatgtt tactacactc ggatatttga tatgtggatt
4441 tcgagtcgtc ttaatgtata gatttgaaga agagctgttt ttacgatccc ttcaggatta
4501 caaaattcaa agtgcgttgc tagtaccaac cctattttca ttcttcgcca aaagcactct
4561 gattgacaaa tacgatttat ctaatttaca cgaaattgct tctgggggcg cacctctttc
4621 gaaagaagtc ggggaagcgg ttgcaaaacg cttccatctt ccagggatac gacaaggata
4681 tgggctcact gagactacat cagctattct gattacaccc gagggggatg ataaaccggg
4741 cgcggtcggt aaagttgttc cattttttga agcgaaggtt gtggatctgg ataccgggaa
4801 aacgctgggc gttaatcaga gaggcgaatt atgtgtcaga ggacctatga ttatgtccgg
4861 ttatgtaaac aatccggaag cgaccaacgc cttgattgac aaggatggat ggctacattc
4921 tggagacata gcttactggg acgaagacga acacttcttc atagttgacc gcttgaagtc
4981 tttaattaaa tacaaaggat atcaggtggc ccccgctgaa ttggaatcga tattgttaca
5041 acaccccaac atcttcgacg cgggcgtggc aggtcttccc gacgatgacg ccggtgaact
5101 tcccgccgcc gttgttgttt tggagcacgg aaagacgatg acggaaaaag agatcgtgga
5161 ttacgtcgcc agtcaagtaa caaccgcgaa aaagttgcgc ggaggagttg tgtttgtgga
5221 cgaagtaccg aaaggtctta ccggaaaact cgacgcaaga aaaatcagag agatcctcat
5281 aaaggccaag aagggcggaa agtccaaatt gtaaaatgta actgtattca gcgatgacga
5341 aattcttagc tattgtaata ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc
5401 agttattggt gcccttaaac gcctggttgc tacgcctgaa taagtgataa taagcggatg
5461 aatggcagaa attcgccgga tctttgtgaa ggaaccttac ttctgtggtg tgacataatt
5521 ggacaaacta cctacagaga tttaaagctc taaggtaaat ataaaatttt taagtgtata
5581 atgtgttaaa ctactgattc taattgtttg tgtattttag attccaacct atggaactga
5641 tgaatgggag cagtggtgga atgcctttaa tgaggaaaac ctgttttgct cagaagaaat
5701 gccatctagt gatgatgagg ctactgctga ctctcaacat tctactcctc caaaaaa
//
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