| 质粒名称: | pAB045a |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 79224 |
| 存储实验室: | David Liu |
|
载体骨架基本信息 |
|
| 载体名称: | ColEI |
| 载体出品公司: | David Liu Lab |
| 载体抗性: | 大观霉素 |
| 空载体大小: | 2992 bp |
| 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: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | PlacZ rpoZ-HA4; PlacIq lacI |
| 物种来源: | Synthetic |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | PlacZ; PlacIq |
| 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: | pAB045a was a gift from David Liu (Addgene plasmid # 79224 ; http://n2t.net/addgene:79224 ; RRID:Addgene_79224) |
| 参考文献部分: | Continuous evolution of Bacillus thuringiensis toxins overcomes insect resistance. Badran AH, Guzov VM, Huai Q, Kemp MM, Vishwanath P, Kain W, Nance AM, Evdokimov A, Moshiri F, Turner KH, Wang P, Malvar T, Liu DR. Nature. 2016 May 5;533(7601):58-63. doi: 10.1038/nature17938. Epub 2016 Apr 27. 10.1038/nature17938 PubMed 27120167 |
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 pAB045a 4309 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Directed Evolution of Protein-Protein Interactions.
ACCESSION .
VERSION .
KEYWORDS pAB045a
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4309)
AUTHORS Badran AH, Guzov VM, Huai Q, Kemp MM, Vishwanath P, Kain W, Nance
AM, Evdokimov A, Moshiri F, Turner KH, Wang P, Malvar T, Liu DR
TITLE Continuous evolution of Bacillus thuringiensis toxins overcomes
insect resistance.
JOURNAL Nature. 2016 May 5;533(7601):58-63. doi: 10.1038/nature17938. Epub
2016 Apr 27.
PUBMED 27120167
REFERENCE 2 (bases 1 to 4309)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4309
/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"
protein_bind 1426..1447
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 1462..1492
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1500..1516
/label=lac operator
/bound_moiety="lac repressor encoded by 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)."
CDS 1555..1827
/codon_start=1
/gene="E. coli rpoZ"
/product="DNA-directed RNA polymerase subunit omega"
/label=RNAP omega
/note="promotes RNA polymerase assembly"
/translation="MARVTVQDAVEKIGNRFDLVLVAARRARQMQVGGKDPLVPEENDK
TTVIALREIEEGLINNQILDVRERQEQQEQEAAELQAVTAIAEGRR"
CDS 1837..1866
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS complement(2251..3042)
/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(3043..3114)
/gene="bla"
/label=AmpR promoter
CDS complement(3144..4226)
/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 4188..4207
/label=LacI-R
/note="LacI, reverse primer"
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 aacaagagga catccggtca
1321 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttgctgagg agacttaggg
1381 accctggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
1441 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
1501 tgtgagcgga taacaatttc acacacctgc aggtgcagta aggaggaaaa aaaaatggca
1561 cgcgtaactg ttcaggacgc tgtagagaaa attggtaacc gttttgacct ggtactggtc
1621 gccgcgcgtc gcgctcgtca gatgcaggta ggcggaaagg atccgctggt accggaagaa
1681 aacgataaaa ccactgtaat cgcgctgcgc gaaatcgaag aaggtctgat caacaaccag
1741 atcctcgacg ttcgcgaacg ccaggaacag caagagcagg aagccgctga attacaagcc
1801 gttaccgcta ttgctgaagg tcgtcgtgcg gccgcggaac aaaagcttat ttctgaagag
1861 gacttgggca gctctgtgag tagcgttccg accaaactgg aagtggttgc agcaaccccg
1921 acgagcctgc tgatttcttg ggatgccccg atgtctagta gctctgtgta ttactatcgt
1981 atcacctacg gtgaaacggg cggtaacagc ccggtgcagg aatttacggt tccgtatagt
2041 agctctaccg cgacgattag tggcctgagc ccgggtgtgg attacaccat cacggtttat
2101 gcatggggcg aagatagcgc gggttacatg ttcatgtatt ctccgattag tatcaattac
2161 cgcacctgct aacttaatta acggcactcc tcagcaaata taatgaccct cttgataacc
2221 caagagggca ttttttaatg cccatggcgt ttatttgccg actaccttgg tgatctcgcc
2281 tttcacgtag tggacaaatt cttccaactg atctgcgcgc gaggccaagc gatcttcttc
2341 ttgtccaaga taagcctgtc tagcttcaag tatgacgggc tgatactggg ccggcaggcg
2401 ctccattgcc cagtcggcag cgacatcctt cggcgcgatt ttgccggtta ctgcgctgta
2461 ccaaatgcgg gacaacgtaa gcactacatt tcgctcatcg ccagcccagt cgggcggcga
2521 gttccatagc gttaaggttt catttagcgc ctcaaataga tcctgttcag gaaccggatc
2581 aaagagttcc tccgccgctg gacctaccaa ggcaacgcta tgttctcttg cttttgtcag
2641 caagatagcc agatcaatgt cgatcgtggc tggctcgaag atacctgcaa gaatgtcatt
2701 gcgctgccat tctccaaatt gcagttcgcg cttagctgga taacgccacg gaatgatgtc
2761 gtcgtgcaca acaatggtga cttctacagc gcggagaatc tcgctctctc caggggaagc
2821 cgaagtttcc aaaaggtcgt tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt
2881 caccgtaacc agcaaatcaa tatcactgtg tggcttcagg ccgccatcca ctgcggagcc
2941 gtacaaatgt acggccagca acgtcggttc gagatggcgc tcgatgacgc caactacctc
3001 tgatagttga gtcgatactt cggcgatcac cgcttccctc atactcttcc tttttcaata
3061 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
3121 gaaaaatatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
3181 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
3241 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
3301 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
3361 atggtggtta atggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
3421 gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
3481 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
3541 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
3601 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
3661 aatgggcccg ctaacagtgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
3721 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
3781 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
3841 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
3901 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
3961 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
4021 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
4081 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
4141 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
4201 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
4261 tatcatgcca taccgcgaaa ggttttgcac cataggccaa ataggccgt
//
