| 质粒名称: | T7-CMVtrans-FFLuc-polyA |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 101156 |
| 存储实验室: | Marcel Bruchez |
|
载体骨架基本信息 |
|
| 载体名称: | Promega L482A |
| 载体出品公司: | Promega |
| 空载体大小: | 2678 bp |
| 完整质粒大小: | 4331 bp |
| 载体修饰: | 荧光素酶AGACCCAAGCTTTCAGATCCGCTAGCGCTACCGGGGATCCagccacc的T7启动子到ATG的序列修饰 |
| 载体类型: | 萤光素酶 |
| 载体抗性: | 氨苄霉素100 μg/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: | Firefly luciferase,FFluc |
| 物种来源: | Photinus pyralis |
| 插入大小: | 1653 bp |
| 启动子: | T7 promoter |
| 5’测序引物 | AATACGCAAACCGCCTCTC |
| 3’测序引物 | GGTGATGTCGGCGATATAGG |
|
文献引用方法 |
|
| 材料和方法部分: | T7-CMVtrans-FFLuc-polyA was a gift from Marcel Bruchez (Addgene plasmid # 101156 ; http://n2t.net/addgene:101156 ; RRID:Addgene_101156) |
| 参考文献部分: | In Vitro Reversible Translation Control Using gammaPNA Probes. Canady TD, Telmer CA, Oyaghire SN, Armitage BA, Bruchez MP. J Am Chem Soc. 2015 Aug 19;137(32):10268-75. doi: 10.1021/jacs.5b05351. Epub 2015 Aug 4. 10.1021/jacs.5b05351 PubMed 26241615 |
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 T7-CMVtrans-FFLuc-polyA 4331 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Template for T7 RNAP production of RNA transcript for expression of
firefly luciferase in a eukaryotic mammalian cell free lysate..
ACCESSION .
VERSION .
KEYWORDS T7-CMVtrans-FFLuc-polyA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4331)
AUTHORS Canady TD, Telmer CA, Oyaghire SN, Armitage BA, Bruchez MP
TITLE In Vitro Reversible Translation Control Using gammaPNA Probes.
JOURNAL J Am Chem Soc. 2015 Aug 19;137(32):10268-75. doi:
10.1021/jacs.5b05351. Epub 2015 Aug 4.
PUBMED 26241615
REFERENCE 2 (bases 1 to 4331)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4331
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 15..32
/label=L4440
/note="L4440 vector, forward primer"
primer_bind 113..132
/label=T7
/note="T7 promoter, forward primer"
promoter 113..131
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 174..183
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 180..1832
/codon_start=1
/gene="luc"
/product="firefly luciferase"
/label=luciferase
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
primer_bind complement(246..264)
/label=LucNrev
/note="Firefly luciferase, reverse primer"
primer_bind complement(1970..1989)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind complement(2203..2222)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2322..2344
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(2382..2400)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 2468..2572
/gene="bla"
/label=AmpR promoter
CDS 2573..3433
/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 complement(2791..2810)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3604..4192
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4093..4112
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
61 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcttatcgaa attaatacga
121 ctcactatag ggagacccaa gctttcagat ccgctagcgc taccggggat ccagccacca
181 tggaagacgc caaaaacata aagaaaggcc cggcgccatt ctatcctcta gaggatggaa
241 ccgctggaga gcaactgcat aaggctatga agagatacgc cctggttcct ggaacaattg
301 cttttacaga tgcacatatc gaggtgaaca tcacgtacgc ggaatacttc gaaatgtccg
361 ttcggttggc agaagctatg aaacgatatg ggctgaatac aaatcacaga atcgtcgtat
421 gcagtgaaaa ctctcttcaa ttctttatgc cggtgttggg cgcgttattt atcggagttg
481 cagttgcgcc cgcgaacgac atttataatg aacgtgaatt gctcaacagt atgaacattt
541 cgcagcctac cgtagtgttt gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa
601 aaaaattacc aataatccag aaaattatta tcatggattc taaaacggat taccagggat
661 ttcagtcgat gtacacgttc gtcacatctc atctacctcc cggttttaat gaatacgatt
721 ttgtaccaga gtcctttgat cgtgacaaaa caattgcact gataatgaat tcctctggat
781 ctactgggtt acctaagggt gtggcccttc cgcatagaac tgcctgcgtc agattctcgc
841 atgccagaga tcctattttt ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg
901 ttccattcca tcacggtttt ggaatgttta ctacactcgg atatttgata tgtggatttc
961 gagtcgtctt aatgtataga tttgaagaag agctgttttt acgatccctt caggattaca
1021 aaattcaaag tgcgttgcta gtaccaaccc tattttcatt cttcgccaaa agcactctga
1081 ttgacaaata cgatttatct aatttacacg aaattgcttc tgggggcgca cctctttcga
1141 aagaagtcgg ggaagcggtt gcaaaacgct tccatcttcc agggatacga caaggatatg
1201 ggctcactga gactacatca gctattctga ttacacccga gggggatgat aaaccgggcg
1261 cggtcggtaa agttgttcca ttttttgaag cgaaggttgt ggatctggat accgggaaaa
1321 cgctgggcgt taatcagaga ggcgaattat gtgtcagagg acctatgatt atgtccggtt
1381 atgtaaacaa tccggaagcg accaacgcct tgattgacaa ggatggatgg ctacattctg
1441 gagacatagc ttactgggac gaagacgaac acttcttcat agttgaccgc ttgaagtctt
1501 taattaaata caaaggatat caggtggccc ccgctgaatt ggaatcgata ttgttacaac
1561 accccaacat cttcgacgcg ggcgtggcag gtcttcccga cgatgacgcc ggtgaacttc
1621 ccgccgccgt tgttgttttg gagcacggaa agacgatgac ggaaaaagag atcgtggatt
1681 acgtcgccag tcaagtaaca accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg
1741 aagtaccgaa aggtcttacc ggaaaactcg acgcaagaaa aatcagagag atcctcataa
1801 aggccaagaa gggcggaaag tccaaattgt aaaatgtaac tgtattcagc gatgacgaaa
1861 ttcttagcta ttgtaatcct ccgagggggc gagctcccaa aaaaaaaaaa aaaaaaaaaa
1921 aaaaaaaacc gaattggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
1981 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
2041 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca
2101 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg
2161 caggagtcgc ataagggaga gcgtcgaatg gtgcactctc agtacaatct gctctgatgc
2221 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
2281 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
2341 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
2401 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
2461 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
2521 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
2581 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
2641 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
2701 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
2761 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
2821 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
2881 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
2941 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
3001 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
3061 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
3121 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
3181 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
3241 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
3301 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
3361 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
3421 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
3481 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
3541 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
3601 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
3661 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
3721 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
3781 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
3841 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
3901 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
3961 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
4021 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
4081 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
4141 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
4201 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
4261 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
4321 tcgccgcagc c
//
