| 质粒名称: | pDawn |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 43796 |
| 存储实验室: | Andreas Moeglich |
|
载体骨架基本信息 |
|
| 载体名称: | pUC |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 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: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | T7term |
|
文献引用方法 |
|
| 材料和方法部分: | pDawn was a gift from Andreas Moeglich (Addgene plasmid # 43796 ; http://n2t.net/addgene:43796 ; RRID:Addgene_43796) |
| 参考文献部分: | From dusk till dawn: one-plasmid systems for light-regulated gene expression. Ohlendorf R, Vidavski RR, Eldar A, Moffat K, Moglich A. J Mol Biol. 2012 Mar 2;416(4):534-42. doi: 10.1016/j.jmb.2012.01.001. Epub 2012 Jan 8. 10.1016/j.jmb.2012.01.001 PubMed 22245580 |
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 pDawn 7213 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pDawn
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7213)
AUTHORS Ohlendorf R, Vidavski RR, Eldar A, Moffat K, Moglich A
TITLE From dusk till dawn: one-plasmid systems for light-regulated gene
expression.
JOURNAL J Mol Biol. 2012 Mar 2;416(4):534-42. doi:
10.1016/j.jmb.2012.01.001. Epub 2012 Jan 8.
PUBMED 22245580
REFERENCE 2 (bases 1 to 7213)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7213
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 26..73
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 69..87
/label=T7 Term
/note="T7 terminator, reverse primer"
CDS complement(140..157)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS complement(205..237)
/codon_start=1
/product="leader peptide from bacteriophage T7 gene 10"
/label=T7 tag (gene 10 leader)
/note="promotes efficient translation in E. coli"
/translation="MASMTGGQQMG"
CDS complement(241..258)
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS complement(268..285)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
RBS 304..326
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
terminator 402..429
/label=T7Te terminator
/note="phage T7 early transcription terminator"
terminator 445..516
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(597..1307)
/codon_start=1
/gene="cI"
/product="phage lambda repressor"
/label=-lambda repressor
/translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
VGKVIASQWPEETFG"
RBS 1314..1325
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
primer_bind complement(1648..1667)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter 1873..1950
/gene="lacI (mutant)"
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
CDS 4508..4699
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
primer_bind complement(4693..4715)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 4801..4943
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 4856..4875
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(4958..4975)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5129..5717)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5209..5228)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind complement(5828..5847)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 5839..6654
/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 complement(5912..5931)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
rep_origin complement(6747..7202)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6884..6905)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 7096..7115
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
181 cgacggagct cgaattcgga tccgacccat ttgctgtcca ccagtcatgc tagccatatg
241 gctgccgcgc ggcaccaggc cgctgctgtg atgatgatga tgatggctgc tgcccatggt
301 atatctcctt cttaaagtta aacaaaatta tttctagagc aaccattatc accgccagag
361 gtaaaatagt caacacgcac ggtgttactc tagtatataa acgcagaaag gcccacccga
421 aggtgagcca gtgtgactct agtagagagc gttcaccgac aaacaacaga taaaacgaaa
481 ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggctctag tagcgatcta
541 cactagcact atcagcgtta ttaagctact aaagcgtagt tttcgtcgtt tgcagcgcca
601 aacgtctctt caggccactg actagcgata actttcccca caacggaaca actctcattg
661 catgggatca ttgggtactg tgggtttagt ggttgtaaaa acacctgacc gctatccctg
721 atcagtttct tgaaggtaaa ctcatcaccc ccaagtctgg ctatgcagaa atcacctggc
781 tcaacagcct gctcagggtc aacgagaatt aacattccgt caggaaagct cggcttggag
841 cctgttggtg cggtcatgga attaccttca acctcaagcc agaatgcaga atcactggct
901 tttttggttg tgcttaccca tctctccgca tcacctttgg taaaggttct aagctcaggt
961 gagaacatcc ctgcctgaac atgagaaaaa acagggtact catactcact tctaagtgac
1021 ggctgcatac taaccgcttc atacatctcg tagatttctc tggcgattga agggctaaat
1081 tcttcaacgc taactttgag aatttttgca agcaatgcgg cgttataagc atttaatgca
1141 ttgatgccat taaataaagc accaacgcct gactgcccca tccccatctt gtctgcgaca
1201 gattcctggg ataagccaag ttcatttttc tttttttcat aaattgcttt aaggcgacgt
1261 gcgtcctcaa gctgctcttg tgttaatggt ttcttttttg tgctcatcta gtatttctcc
1321 tcttttctag actccgttgt gatgacgcat tggtacgcgg tatcgggagg ttcgaaaatt
1381 tcgagcgata tcttaagggg ggtgccttac gtagaacccc gtaggtcatg cccgaggccg
1441 gtcctggatg gcgcggcgga tacgcttgag caggttttcg tcgagaagcg gcttcaaaac
1501 cacgtctttt acgccggcct cggcggcccg ggtcgagatg ttttcgtccg gatagccggt
1561 gatcaggatc acgggcgtag atctcgatcc tctacgccgg acgcatcgtg gccggcatca
1621 ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc
1681 gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg
1741 tggccggggg actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc
1801 tcaacggcct caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc
1861 gtcgagatcc cggacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc
1921 ccggaagaga gtcaattgag ggtggtgaat gtggctagtt ttcaatcatt tgggatacca
1981 ggacagctgg aagtcatcaa aaaagcactt gatcacgtgc gagtcggtgt ggtaattaca
2041 gatcccgcac ttgaagataa tcctattgtc tacgtaaatc aaggctttgt tcaaatgacc
2101 ggctacgaga ccgaggaaat tttaggaaag aactgtcgct tcttacaggg gaaacacaca
2161 gatcctgcag aagtggacaa catcagaacc gctttacaaa ataaagaacc ggtcaccgtt
2221 cagatccaaa actacaaaaa agacggaacg atgttctgga atgaattaaa tattgatcca
2281 atggaaatag aggataaaac gtattttgtc ggtattcaga atgatatcac cgagcaccag
2341 cagacccagg cgcgcctcca ggaactgcaa tccgagctcg tccacgtctc caggctgagc
2401 gccatgggcg aaatggcgtc cgcgctcgcg cacgagctca accagccgct ggcggcgatc
2461 agcaactaca tgaagggctc gcggcggctg cttgccggca gcagtgatcc gaacacaccg
2521 aaggtcgaaa gcgccctgga ccgcgccgcc gagcaggcgc tgcgcgccgg ccagatcatc
2581 cggcgcctgc gcgacttcgt tgcccgcggc gaatcggaga agcgggtcga gagtctctcc
2641 aagctgatcg aggaggccgg cgcgctcggg cttgccggcg cgcgcgagca gaacgtgcag
2701 ctccgcttca gtctcgatcc gggcgccgat ctcgttctcg ccgaccgggt gcagatccag
2761 caggtcctgg tcaacctgtt ccgcaacgcg ctggaagcga tggctcagtc gcagcgacgc
2821 gagctcgtcg tcaccaacac ccccgccgcc gacgacatga tcgaggtcga agtgtccgac
2881 accggcagcg gtttccagga cgacgtcatt ccgaacctgt ttcagacttt cttcaccacc
2941 aaggacaccg gcatgggcgt gggactgtcc atcagccgct cgatcatcga agctcacggc
3001 gggcgcatgt gggccgagag caacgcatcg ggcggggcga ccttccgctt caccctcccg
3061 gcagccgacg agatgatagg aggtctagca tgacgaccaa gggacatatc tacgtcatcg
3121 acgacgacgc ggcgatgcgg gattcgctga atttcctgct ggattctgcc ggcttcggcg
3181 tcacgctgtt tgacgacgcg caagcctttc tcgacgccct gccgggtctc tccttcggct
3241 gcgtcgtctc cgacgtgcgc atgccgggcc ttgacggcat cgagctgttg aagcggatga
3301 aggcgcagca aagccccttt ccgatcctca tcatgaccgg tcacggcgac gtgccgctcg
3361 cggtcgaggc gatgaagtta ggggcggtgg actttctgga aaagcctttc gaggacgacc
3421 gcctcaccgc catgatcgaa tcggcgatcc gccaggccga gccggccgcc aagagcgagg
3481 ccgtcgcgca ggatatcgcc gcccgcgtcg cctcgttgag ccccagggag cgccaggtca
3541 tggaagggct gatcgccggc ctttccaaca agctgatcgc ccgcgagtac gacatcagcc
3601 cgcgcaccat cgaggtgtat cgggccaacg tcatgaccaa gatgcaggcc aacagccttt
3661 cggagctggt tcgcctcgcg atgcgcgccg gcatgctcaa cgattgacaa ttgatgtaag
3721 ttagctcact cattaggcac cgggatctcg accgatgccc ttgagagcct tcaacccagt
3781 cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt
3841 tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg gcgaggaccg
3901 ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc
3961 cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga agcaggccat
4021 tatcgccggc atggcggccc cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg
4081 gctaggctgg cggggttgcc ttactggtta gcagaatgaa tcaccgatac gcgagcgaac
4141 gtgaagcgac tgctgctgca aaacgtctgc gacctgagca acaacatgaa tggtcttcgg
4201 tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca ttatgttccg
4261 gatctgcatc gcaggatgct gctggctacc ctgtggaaca cctacatctg tattaacgaa
4321 gcgctggcat tgaccctgag tgatttttct ctggtcccgc cgcatccata ccgccagttg
4381 tttaccctca caacgttcca gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag
4441 catcctctct cgtttcatcg gtatcattac ccccatgaac agaaatcccc cttacacgga
4501 ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca
4561 gacattaacg cttctggaga aactcaacga gctggacgcg gatgaacagg cagacatctg
4621 tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc gtttcggtga
4681 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
4741 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
4801 cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca
4861 tcagagcaga ttgtactgag agtgcaccat atatgcggtg tgaaataccg cacagatgcg
4921 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct
4981 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
5041 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
5101 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
5161 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
5221 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
5281 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
5341 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
5401 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
5461 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
5521 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
5581 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
5641 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
5701 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
5761 acgaaaactc acgttaaggg attttggtca tgaacaataa aactgtctgc ttacataaac
5821 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgctc taggccgcga
5881 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
5941 caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg
6001 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg
6061 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
6121 tggttactca ccactgcgat ccccgggaaa acagcattcc aggtattaga agaatatcct
6181 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
6241 cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca
6301 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct
6361 gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc
6421 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
6481 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
6541 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
6601 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaagaatta
6661 attcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
6721 cacatttccc cgaaaagtgc cacctgaaat tgtaaacgtt aatattttgt taaaattcgc
6781 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
6841 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
6901 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
6961 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
7021 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
7081 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
7141 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
7201 gtcccattcg cca
//
