| 质粒名称: | pSBL_DK193 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 156424 |
| 存储实验室: | Stephen Brohawn |
|
载体骨架基本信息 |
|
| 载体名称: | Derived from pACEBAC1 |
| 载体出品公司: | Geneva Biotech |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 昆虫表达 |
| 筛选标记: | -- |
| 载体抗性: | 庆大霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | SARS-CoV-2 3a protein (Q57H) |
| 物种来源: | SARS-CoV-2 |
| 插入大小: | -- |
| 基因突变: | Q57H |
| GenBank编号 | -- |
| 启动子: | Polyhedrin |
| 标签/融合蛋白: | sfGFP His（主干上的C终端） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | XhoI(未破坏) |
| 3’克隆位点: | EcoRI(未破坏) |
| 5’测序引物 | pFASTBAC Forward |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pSBL_DK193 was a gift from Stephen Brohawn (Addgene plasmid # 156424 ; http://n2t.net/addgene:156424 ; RRID:Addgene_156424) |
| 参考文献部分: | Cryo-EM structure of the SARS-CoV-2 3a ion channel in lipid nanodiscs. Kern DM, Sorum B, Hoel CM, Sridharan S, Remis JP, Toso DB, Brohawn SG. BioRxiv 2020 10.1101/2020.06.17.156554 |
买家导航
pSBLDK193载体基本信息
pSBLDK193载体图谱质粒图谱和多克隆位点信息
pSBLDK193载体简介
pSBLDK193载体序列
LOCUS pSBL_DK193 4478 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Insect cell expression of 3a(Q57H)-sfGFP, needs to be introduced
into baculoviral bacmid.
ACCESSION .
VERSION .
KEYWORDS pSBL_DK193
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4478)
AUTHORS Kern DM, Sorum B, Hoel CM, Sridharan S, Remis JP, Toso DB, Brohawn
SG
TITLE Cryo-EM structure of the SARS-CoV-2 3a ion channel in lipid
nanodiscs
JOURNAL BioRxiv 2020
REFERENCE 2 (bases 1 to 4478)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4478
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 395..414
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
mobile_element 629..852
/mobile_element_type="transposon:Tn7"
/note="mini-Tn7 element (right end of the Tn7 transposon)"
CDS complement(919..1452)
/codon_start=1
/gene="aacC1"
/product="gentamycin acetyltransferase"
/label=GmR
/note="confers resistance to gentamycin"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(1641..1669)
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
protein_bind 1740..1773
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
promoter 1850..1941
/gene="polh from Autographa californica"
/label=polyhedrin promoter
/note="promoter for the baculovirus polyhedrin gene"
primer_bind 1866..1883
/label=Polyhedrin-F
/note="Polyhedrin promoter, forward primer"
regulatory 1989..1998
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2829..2852
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
CDS 2898..3611
/codon_start=1
/product="GFP variant that folds robustly even when fused
to poorly folded proteins (Pedelacq et al., 2006)"
/label=superfolder GFP
/translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
CDS 3621..3641
/codon_start=1
/product="6xHis affinity tag"
/label=7xHis
/translation="HHHHHHH"
polyA_signal 3774..3908
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3811..3830)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3865..3884
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
mobile_element 4029..4194
/mobile_element_type="transposon:Tn7"
/note="mini-Tn7 element (left end of the Tn7 transposon)"
rep_origin 4384..494
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
61 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
121 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
181 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
241 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
301 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
361 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
421 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
481 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
541 cttttgctca catgttcttt cctgcgttat cccctgattg acttgggtcg ctcttcctgt
601 ggatgcgcag atgccctgcg taagcgggtg tgggcggaca ataaagtctt aaactgaaca
661 aaatagatct aaactatgac aataaagtct taaactagac agaatagttg taaactgaaa
721 tcagtccagt tatgctgtga aaaagcatac tggacttttg ttatggctaa agcaaactct
781 tcattttctg aagtgcaaat tgcccgtcgt attaaagagg ggcgtggcca agggcatgta
841 aagactatat tcgcggcgtt gtgacaattt accgaacaac tccgcggccg ggaagccgat
901 ctcggcttga acgaattgtt aggtggcggt acttgggtcg atatcaaagt gcatcacttc
961 ttcccgtatg cccaactttg tatagagagc cactgcggga tcgtcaccgt aatctgcttg
1021 cacgtagatc acataagcac caagcgcgtt ggcctcatgc ttgaggagat tgatgagcgc
1081 ggtggcaatg ccctgcctcc ggtgctcgcc ggagactgcg agatcataga tatagatctc
1141 actacgcggc tgctcaaact tgggcagaac gtaagccgcg agagcgccaa caaccgcttc
1201 ttggtcgaag gcagcaagcg cgatgaatgt cttactacgg agcaagttcc cgaggtaatc
1261 ggagtccggc tgatgttggg agtaggtggc tacgtctccg aactcacgac cgaaaagatc
1321 aagagcagcc cgcatggatt tgacttggtc agggccgagc ctacatgtgc gaatgatgcc
1381 catacttgag ccacctaact ttgttttagg gcgactgccc tgctgcgtaa catcgttgct
1441 gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccacggcg taacgcgctt
1501 gctgcttgga tgcccgaggc atagactgta caaaaaaaca gtcataacaa gccatgaaaa
1561 ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt tctggaccag ttgcgtgagc
1621 gcatacgcta cttgcattac agtttacgaa ccgaacaggc ttatgtcaac tgggttcgtg
1681 ccttcatccg tttccacggt gtgcgtcacc cggcaacctt gggcagcagc gaagtcgcca
1741 taacttcgta tagcatacat tatacgaagt tatctgtaac tataacggtc ctaaggtagc
1801 gagtttaaac actagtatcg attcgcgacc tactccggaa tattaataga tcatggagat
1861 aattaaaatg ataaccatct cgcaaataaa taagtatttt actgttttcg taacagtttt
1921 gtaataaaaa aacctataaa tattccggat tattcatacc gtcccaccat cgggcgcgga
1981 tccctcgagc caccatggat ctctttatgc gcatttttac gatcggtaca gttacgttga
2041 agcaaggtga aatcaaagac gccacgccgt ccgattttgt tcgcgctacg gctacaattc
2101 caatccaagc gagcttgcca ttcggttggt tgatagttgg tgttgcgttg cttgcagtct
2161 ttcactcagc ttctaagata atcactctga agaagcgttg gcaactggcg ctctcaaaag
2221 gcgttcactt cgtgtgcaac ctcctgttgc tctttgtcac cgtttactca catctcttgc
2281 ttgttgctgc tggcctggaa gctcccttcc tttacctgta cgctctggtc tattttctgc
2341 agagcataaa ttttgtcagg ataataatga gactgtggct ttgttggaaa tgccgctcca
2401 agaacccgct tttgtacgat gccaattact tcctgtgttg gcatacgaat tgctacgact
2461 actgtattcc gtataatagc gtcacttctt caatcgtgat aacgtccggt gacggtacaa
2521 catcacctat ttctgaacac gattaccaaa taggtggcta caccgagaaa tgggagagtg
2581 gagtcaaaga ttgtgtagtg ctgcactcat attttacctc ggattattat caactctact
2641 caacacagtt gtctactgat acaggagttg agcacgtaac atttttcatt tacaataaaa
2701 ttgtggacga acccgaggag catgtacaga ttcatactat cgacggtagc agcggtgtcg
2761 tgaaccccgt tatggagcca atttacgacg aacccaccac gacaacatcc gtccccctgt
2821 cgaattcttt ggaagttttg tttcaaggtc cgtctagagg tggatctgga gctgcagctg
2881 gatctggttc tggttctatg cgtaaaggcg aagagctgtt cactggtgtc gtccctattc
2941 tggtggaact ggatggtgat gtcaacggtc ataagttttc cgtgcgtggc gagggtgaag
3001 gtgacgcaac taatggtaaa ctgacgctga agttcatctg tactactggt aaactgccgg
3061 taccttggcc gactctggta acgacgctga cttatggtgt tcagtgcttt gctcgttatc
3121 cggaccatat gaagcagcat gacttcttca agtccgccat gccggaaggc tatgtgcagg
3181 aacgcacgat ttcctttaag gatgacggca cgtacaaaac gcgtgcggaa gtgaaatttg
3241 aaggcgatac cctggtaaac cgcattgagc tgaaaggcat tgactttaaa gaagacggca
3301 atatcctggg ccataagctg gaatacaatt ttaacagcca caatgtttac atcaccgccg
3361 ataaacaaaa aaatggcatt aaagcgaatt ttaaaattcg ccacaacgtg gaggatggca
3421 gcgtgcagct ggctgatcac taccagcaaa acactccaat cggtgatggt cctgttctgc
3481 tgccagacaa tcactatctg agcacgcaaa gcgttctgtc taaagatccg aacgagaaac
3541 gcgatcatat ggttctgctg gagttcgtaa ccgcagcggg catcacgcat ggtatggatg
3601 aactgtacaa aggcagcagc catcatcatc atcatcacca ctgaaagctt gtcgagaagt
3661 actagaggat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
3721 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
3781 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
3841 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
3901 tctggatctg atcactgctt gagcctagaa gatccggctg ctaacaaagc ccgaaaggaa
3961 gctgagttgg ctgctgccac cgctgagcaa taactatcat aacccctagg gtatacccat
4021 ctaattggaa ccagataagt gaaatctagt tccaaactat tttgtcattt ttaattttcg
4081 tattagctta cgacgctaca cccagttccc atctattttg tcactcttcc ctaaataatc
4141 cttaaaaact ccatttccac ccctcccagt tcccaactat tttgtccgcc cacaaccggt
4201 tgacttgggt caactgtcag accaagttta ctcatatata ctttagattg atttaaaact
4261 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
4321 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
4381 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
4441 accagcggtg gtttgtttgc cggatcaaga gctaccaa
//
