| 质粒名称: | pDM_noro_A2_lacZA |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 118819 |
| 存储实验室: | Alexander Green |
|
载体骨架基本信息 |
|
| 载体名称: | pCOLAduet |
| 空载体大小: | 4038 bp |
| 完整质粒大小: | 3562bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | Kanamycin, 50 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: | Norovirus toehold switch A2 |
| 物种来源: | Synthetic |
| 插入大小: | 346 bp |
| 克隆方法: | Gibson Cloning |
| 5’克隆位点: | AgeI(未破坏) |
| 3’克隆位点: | NotI (未破坏) |
| 5’测序引物 | CGTTACTGGTTTCACATTCACCACCC |
|
文献引用方法 |
|
| 材料和方法部分: | pDM_noro_A2_lacZA was a gift from Alexander Green (Addgene plasmid # 118819 ; http://n2t.net/addgene:118819 ; RRID:Addgene_118819) |
| 参考文献部分: | Low-cost detection of norovirus using paper-based cell-free systems and synbody-based viral enrichment. Ma D, Shen L, Wu K, Diehnelt CW, Green AA. Synth Biol (Oxf). 2018;3(1):ysy018. doi: 10.1093/synbio/ysy018. Epub 2018 Sep 19. 10.1093/synbio/ysy018 PubMed 30370338 |
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 pDM_noro_A2_lacZA 3562 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION T7 RNAP-driven expression of norovirus antisense orientation toehold
switches (A2) with a lacZ alpha subunit reporter..
ACCESSION .
VERSION .
KEYWORDS pDM_noro_A2_lacZA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3562)
AUTHORS Ma D, Shen L, Wu K, Diehnelt CW, Green AA
TITLE Low-cost detection of norovirus using paper-based cell-free systems
and synbody-based viral enrichment.
JOURNAL Synth Biol (Oxf). 2018;3(1):ysy018. doi: 10.1093/synbio/ysy018. Epub
2018 Sep 19.
PUBMED 30370338
REFERENCE 2 (bases 1 to 3562)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..3562
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 1..20
/label=T7
/note="T7 promoter, forward primer"
promoter 1..19
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 126..299
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
DRPSQQLRSLNGE"
primer_bind complement(146..163)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(146..162)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(155..177)
/label=M13/pUC Forward
/note="In lacZ gene"
terminator 309..355
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(585..1400)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 1308..1327
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
promoter complement(1401..1492)
/gene="bla"
/label=AmpR promoter
rep_origin complement(1510..2145)
/direction=LEFT
/label=ColA ori
/note="Plasmids containing the ColA origin of replication
can be propagated in E. coli cells that contain additional
plasmids with compatible origins."
primer_bind complement(1655..1674)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2343..3425)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind 3387..3406
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(3426..3503)
/gene="lacI"
/label=lacI promoter
ORIGIN
1 taatacgact cactataggg tgggagggcg atcgcaatct ggctcccagt tttgtggact
61 ttagaacaga ggagataaag atgacaaaac tgggaacctg gcggcagcgc aagaagatgc
121 gtaaaatgac catgattacg gattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
181 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
241 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
301 ggtaataata gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
361 acctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa
421 ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga caagctgacg
481 accgggtctc cgcaagtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
541 ttttctaaat acattcaaat atgtatccgc tcatgaatta attcttagaa aaactcatcg
601 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
661 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
721 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
781 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
841 ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
901 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
961 aatacgcggt cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
1021 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
1081 aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata
1141 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
1201 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
1261 ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
1321 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga gcaagacgtt
1381 tcccgttgaa tatggctcat actcttcctt tttcaatatt attgaagcat ttatcagggt
1441 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggcatg
1501 ctagcgcaga aacgtcctag aagatgccag gaggatactt agcagagaga caataaggcc
1561 ggagcgaagc cgtttttcca taggctccgc ccccctgacg aacatcacga aatctgacgc
1621 tcaaatcagt ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgat
1681 ggctccctct tgcgctctcc tgttcccgtc ctgcggcgtc cgtgttgtgg tggaggcttt
1741 acccaaatca ccacgtcccg ttccgtgtag acagttcgct ccaagctggg ctgtgtgcaa
1801 gaaccccccg ttcagcccga ctgctgcgcc ttatccggta actatcatct tgagtccaac
1861 ccggaaagac acgacaaaac gccactggca gcagccattg gtaactgaga attagtggat
1921 ttagatatcg agagtcttga agtggtggcc taacagaggc tacactgaaa ggacagtatt
1981 tggtatctgc gctccactaa agccagttac caggttaagc agttccccaa ctgacttaac
2041 cttcgatcaa accgcctccc caggcggttt tttcgtttac agagcaggag attacgacga
2101 tcgtaaaagg atctcaagaa gatcctttac ggattcccga caccatcact ctagatttca
2161 gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc ccatacgata taagttgtaa
2221 ttctcatgtt agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
2281 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc
2341 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
2401 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt
2461 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg
2521 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata
2581 taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatgtccgc accaacgcgc
2641 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc
2701 atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg
2761 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta
2821 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg
2881 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg
2941 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca
3001 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc
3061 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg
3121 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc
3181 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc
3241 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc
3301 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg
3361 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
3421 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag
3481 gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgtt gaaagtgacc
3541 accgctcacg tgttgtgtgc gc
//
