| 质粒名称: | pDB023f |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 99217 |
| 存储实验室: | David Liu |
|
载体骨架基本信息 |
|
| 载体名称: | ColE1 |
| 载体出品公司: | David Liu Lab |
| 载体抗性: | 大观霉素 |
| 空载体大小: | 2729 bp |
| 完整质粒大小: | 5624 bp |
| 载体修饰: | -- |
| 载体类型: | -- |
| 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: | Ppsp [SD8] T7RNAP(S12*,S203*) |
| 物种来源: | Synthetic |
| 插入大小: | 2895 bp |
| 基因突变: | amber codons at positions 12 and 203 of T7 RNA polymerase |
| GenBank编号 | -- |
| 启动子 | Ppsp |
| 标签/融合蛋白: | -- |
| 克隆方法: | Ligation Independent克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pDB023f was a gift from David Liu (Addgene plasmid # 99217 ; http://n2t.net/addgene:99217 ; RRID:Addgene_99217) |
| 参考文献部分: | Continuous directed evolution of aminoacyl-tRNA synthetases. Bryson DI, Fan C, Guo LT, Miller C, Soll D, Liu DR. Nat Chem Biol. 2017 Oct 16. doi: 10.1038/nchembio.2474. 10.1038/nchembio.2474 PubMed 29035361 |
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 pDB023f 5624 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Complementary plasmid for aaRS PACE; amber codons in T7 RNAP.
ACCESSION .
VERSION .
KEYWORDS pDB023f
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5624)
AUTHORS Bryson DI, Fan C, Guo LT, Miller C, Soll D, Liu DR
TITLE Continuous directed evolution of aminoacyl-tRNA synthetases.
JOURNAL Nat Chem Biol. 2017 Oct 16. doi: 10.1038/nchembio.2474.
PUBMED 29035361
REFERENCE 2 (bases 1 to 5624)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5624
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 18..606
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 507..526
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 760..777
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 792..932
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(858..877)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 1018..1040
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(1034..1225)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
primer_bind complement(1368..1385)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 1538..1624
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 1716..1743
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
CDS complement(4718..5509)
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
promoter complement(5510..5581)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ccaggccaaa taggccgttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
61 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc acctcttttt
121 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
181 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
241 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
301 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
361 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
421 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
481 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
541 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
601 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
661 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
721 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
781 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
841 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
901 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
961 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
1021 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
1081 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
1141 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
1201 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
1261 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
1321 aagaggacat ccggtggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt
1381 aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg
1441 gtcccacctg accccatgcc gaactcagaa gtgaaactcc gtagcgccga tggtagtgtg
1501 gggtctcccc atgcgagagt agggaactgc cagtcatcaa ataaaacgaa aggctcagtc
1561 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
1621 aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg
1681 acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct
1741 ttttgcgttt ctacaaactc gctgaggatt ctagagcaca gcttgatgaa attcgccact
1801 tgttagtgta attcgctaac tcatcctggc atgttgctgt tgattcttca atcagatctt
1861 tataaatcaa aaagataaaa aattggcacg caaattgtat taacagttca gcaggacaat
1921 cctgaacgca gaaatcaaga ggacaacatt aagaaaagtg aaatgaatag ttaaggagga
1981 aaaaaaaatg aacacgatta acatcgctaa gaacgacttc taggacatcg aactggctgc
2041 tatcccgttc aacactctgg ctgaccatta cggtgagcgt ttagctcgcg aacagttggc
2101 ccttgagcat gagtcttacg agatgggtga agcacgcttc cgcaagatgt ttgagcgtca
2161 acttaaagct ggtgaggttg cggataacgc tgccgccaag cctctcatca ctaccctact
2221 ccctaagatg attgcacgca tcaacgactg gtttgaggaa gtgaaagcta agcgcggcaa
2281 gcgcccgaca gccttccagt tcctgcaaga aatcaagccg gaagccgtag cgtacatcac
2341 cattaagacc actctggctt gcctaaccag tgctgacaat acaaccgttc aggctgtagc
2401 aagcgcaatc ggtcgggcca ttgaggacga ggctcgcttc ggtcgtatcc gtgaccttga
2461 agctaagcac ttcaagaaaa acgttgagga acaactcaac aagcgcgtag ggcacgtcta
2521 caagaaagca tttatgcaag ttgtcgaggc tgacatgctc tctaagggtc tactcggtgg
2581 cgaggcgtgg tcttagtggc ataaggaaga ctctattcat gtaggagtac gctgcatcga
2641 gatgctcatt gagtcaaccg gaatggttag cttacaccgc caaaatgctg gcgtagtagg
2701 tcaagactct gagactatcg aactcgcacc tgaatacgct gaggctatcg caacccgtgc
2761 aggtgcgctg gctggcatct ctccgatgtt ccaaccttgc gtagttcctc ctaagccgtg
2821 gactggcatt actggtggtg gctattgggc taacggtcgt cgtcctctgg cgctggtgcg
2881 tactcacagt aagaaagcac tgatgcgcta cgaagacgtt tacatgcctg aggtgtacaa
2941 agcgattaac attgcgcaaa acaccgcatg gaaaatcaac aagaaagtcc tagcggtcgc
3001 caacgtaatc accaagtgga agcattgtcc ggtcgaggac atccctgcga ttgagcgtga
3061 agaactcccg atgaaaccgg aagacatcga catgaatcct gaggctctca ccgcgtggaa
3121 acgtgctgcc gctgctgtgt accgcaagga caaggctcgc aagtctcgcc gtatcagcct
3181 tgagttcatg cttgagcaag ccaataagtt tgctaaccat aaggccatct ggttccctta
3241 caacatggac tggcgcggtc gtgtttacgc tgtgtcaatg ttcaacccgc aaggtaacga
3301 tatgaccaaa ggactgctta cgctggcgaa aggtaaacca atcggtaagg aaggttacta
3361 ctggctgaaa atccacggtg caaactgtgc gggtgtcgat aaggttccgt tccctgagcg
3421 catcaagttc attgaggaaa accacgagaa catcatggct tgcgctaagt ctccactgga
3481 gaacacttgg tgggctgagc aagattctcc gttctgcttc cttgcgttct gctttgagta
3541 cgctggggta cagcaccacg gcctgagcta taactgctcc cttccgctgg cgtttgacgg
3601 gtcttgctct ggcatccagc acttctccgc gatgctccga gatgaggtag gtggtcgcgc
3661 ggttaacttg cttcctagtg aaaccgttca ggacatctac gggattgttg ctaagaaagt
3721 caacgagatt ctacaagcag acgcaatcaa tgggaccgat aacgaagtag ttaccgtgac
3781 cgatgagaac actggtgaaa tctctgagaa agtcaagctg ggcactaagg cactggctgg
3841 tcaatggctg gcttacggtg ttactcgcag tgtgactaag cgttcagtca tgacgctggc
3901 ttacgggtcc aaagagttcg gcttccgtca acaagtgctg gaagatacca ttcagccagc
3961 tattgattcc ggcaagggtc tgatgttcac tcagccgaat caggctgctg gatacatggc
4021 taagctgatt tgggaatctg tgagcgtgac ggtggtagct gcggttgaag caatgaactg
4081 gcttaagtct gctgctaagc tgctggctgc tgaggtcaaa gataagaaga ctggagagat
4141 tcttcgcaag cgttgcgctg tgcattgggt aactcctgat ggtttccctg tgtggcagga
4201 atacaagaag cctattcaga cgcgcttgaa cctgatgttc ctcggtcagt tccgcttaca
4261 gcctaccatt aacaccaaca aagatagcga gattgatgca cacaaacagg agtctggtat
4321 cgctcctaac tttgtacaca gccaagacgg tagccacctt cgtaagactg tagtgtgggc
4381 acacgagaag tacggaatcg aatcttttgc actgattcac gactccttcg gtaccattcc
4441 ggctgacgct gcgaacctgt tcaaagcagt gcgcgaaact atggttgaca catatgagtc
4501 ttgtgatgta ctggctgatt tctacgacca gttcgctgac cagttgcacg agtctcaatt
4561 ggacaaaatg ccagcacttc cggctaaagg taacttgaac ctccgtgaca tcttagagtc
4621 ggacttcgcg ttcgcgtaac ttaattaacg gcactcctca gcaaatataa tgaccctctt
4681 gataacccaa gagggcattt tttaatgccc atggcgttta tttgccgact accttggtga
4741 tctcgccttt cacgtagtgg acaaattctt ccaactgatc tgcgcgcgag gccaagcgat
4801 cttcttcttg tccaagataa gcctgtctag cttcaagtat gacgggctga tactgggccg
4861 gcaggcgctc cattgcccag tcggcagcga catccttcgg cgcgattttg ccggttactg
4921 cgctgtacca aatgcgggac aacgtaagca ctacatttcg ctcatcgcca gcccagtcgg
4981 gcggcgagtt ccatagcgtt aaggtttcat ttagcgcctc aaatagatcc tgttcaggaa
5041 ccggatcaaa gagttcctcc gccgctggac ctaccaaggc aacgctatgt tctcttgctt
5101 ttgtcagcaa gatagccaga tcaatgtcga tcgtggctgg ctcgaagata cctgcaagaa
5161 tgtcattgcg ctgccattct ccaaattgca gttcgcgctt agctggataa cgccacggaa
5221 tgatgtcgtc gtgcacaaca atggtgactt ctacagcgcg gagaatctcg ctctctccag
5281 gggaagccga agtttccaaa aggtcgttga tcaaagctcg ccgcgttgtt tcatcaagcc
5341 ttacggtcac cgtaaccagc aaatcaatat cactgtgtgg cttcaggccg ccatccactg
5401 cggagccgta caaatgtacg gccagcaacg tcggttcgag atggcgctcg atgacgccaa
5461 ctacctctga tagttgagtc gatacttcgg cgatcaccgc ttccctcata ctcttccttt
5521 ttcaatatta ttgaagcatt tatcagggtt attgtttcat gagcgtatac atatttgaat
5581 gtatttagaa aaatattaat tgcgttgcgc tcactgcccg cttt
//
