| 质粒名称: | pDB016 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 99220 |
| 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: | 2797 bp |
| 完整质粒大小: | 3981 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 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: | PT7 [SD8] gIII-neg |
| 物种来源: | Synthetic |
| 插入大小: | 1184 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | PT7 |
| 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: | pDB016 was a gift from David Liu (Addgene plasmid # 99220 ; http://n2t.net/addgene:99220 ; RRID:Addgene_99220) |
| 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 pDB016 3979 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Accessory plasmid for aaRS PACE; negative selection.
ACCESSION .
VERSION .
KEYWORDS pDB016
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3979)
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 3979)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3979
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 1..589
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 490..509
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 743..760
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 775..915
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(841..860)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 1001..1023
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(1017..1208)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
primer_bind 1826..1845
/label=T7
/note="T7 promoter, forward primer"
promoter 1826..1844
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1948..1965
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS complement(3056..3847)
/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(3848..3919)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
61 agcggtggtt tgtttgccgg atcaagagct accacctctt tttccgaagg taactggctt
121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
781 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
841 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
901 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
961 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
1021 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
1081 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
1141 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
1201 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
1261 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacagaggac atccggtaag
1321 gccaataggg gttcataagg tacggcccgt actgaacccg ggtaggctgc ttgagccagt
1381 gagcgattgc tggcctagat gaatgactgt ccacgacaga acccggctta tcggtcagtt
1441 tcacctgatt tacgtaaaaa cccgcttcgg cgggtttttg cttttggagg ggcagaaaga
1501 tgaatgactg tccacgacgc tatacccaaa agaaagcggc ttatcggtca gtttcacctg
1561 gtttacgtaa aaacccgctt cggcgggttt ttgcttttgg aggggcagaa agatgaatga
1621 ctgtccacga cactataccc aaaagaaagc ggcttatcgg tcagtttcac ctgttttacg
1681 taaaaacccg cttcggcggg tttttacttt tggaggggca gaaagatgaa tgactgtcca
1741 cgacactata cccaaaagaa agcggcttat cggtcagttt tacctgatgt acgtaataaa
1801 ccgttccggc gggtttggga caaattaata cgactcacta tagggtgaaa gacctgcagg
1861 tgcagtaagg aggaaaaaaa aatgaaaaaa ttattattcg caattccttt agttgttcct
1921 ttctattctc actccgctga aactgttcat caccatcacc atcacgctga aactgttgaa
1981 agttgtttag caaaacccca tacagaaaat tcatttacta acgtctggaa agacgacaaa
2041 actttagatc gttacgctaa ctatgagggc tgtctgtgga atgctacagg cgttgtagtt
2101 tgtactggtg acgaaactca gtgttacggt acatgggttc ctattgggct tgctatccct
2161 gaaaatgagg gtggtggctc tgagggtggc ggttctgagg gtggcggttc tgagggtggc
2221 ggtactaaac ctcctgagta cggtgataca cctattccgg gctatactta tatcaaccct
2281 ctcgacggca cttatccgcc tggtactgag caaaaccccg ctaatcctaa tccttctctt
2341 gaggagtctc agcctcttaa tactttcatg tttcagaata ataggttccg aaataggcag
2401 ggggcattaa ctgtttatac gggcactgtt actcaaggca ctgaccccgt taaaacttat
2461 taccagtaca ctcctgtatc atcaaaagcc atgtatgacg cttactggaa cggtaaattc
2521 agagactgcg ctttccattc tggctttaat gaggatccat tcgtttgtga atatcaaggc
2581 caatcgtctg acctgcctca acctcctgtc aatgctggcg gcggctctgg tggtggttct
2641 ggtggcggct ctgagggtgg tggctctgag ggtggcggtt ctgagggtgg cggctctgag
2701 ggaggcggtt ccggtggtgg ctcttcccaa atggctcaag tcggtgacgg tgataattca
2761 cctttaatga ataatttccg tcaatattta ccttccctcc ctcaatcggt tgaatgtcgc
2821 ccttttgtct ttggcgctgg taaaccttac gagttcagta tcgactgcga taagatcaac
2881 ctgttccgcg gtgtctttgc gtttctttta tatgttgcca cctttatgta tgtattttct
2941 acgtttgcta acatactgcg taataaggag tcttaaactt aattaacggc actcctcagc
3001 aaatataatg accctcttga taacccaaga gggcattttt taatgcccat ggcgtttatt
3061 tgccgactac cttggtgatc tcgcctttca cgtagtggac aaattcttcc aactgatctg
3121 cgcgcgaggc caagcgatct tcttcttgtc caagataagc ctgtctagct tcaagtatga
3181 cgggctgata ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg
3241 cgattttgcc ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct
3301 catcgccagc ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa
3361 atagatcctg ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa
3421 cgctatgttc tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct
3481 cgaagatacc tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag
3541 ctggataacg ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga
3601 gaatctcgct ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc
3661 gcgttgtttc atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct
3721 tcaggccgcc atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat
3781 ggcgctcgat gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt
3841 ccctcatact cttccttttt caatattatt gaagcattta tcagggttat tgtttcatga
3901 gcgtatacat atttgaatgt atttagaaaa atattaattg cgttgcgctc actgcccgct
3961 ttccaggcca aataggccg
//
