| 质粒名称: | pAB029f |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 79220 |
| 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 lambda cI-SH2ABL1; 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: | pAB029f was a gift from David Liu (Addgene plasmid # 79220 ; http://n2t.net/addgene:79220 ; RRID:Addgene_79220) |
| 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 pAB029f 4746 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Directed Evolution of Protein-Protein Interactions.
ACCESSION .
VERSION .
KEYWORDS pAB029f
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4746)
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 4746)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4746
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 489..508
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 742..759
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 774..914
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(840..859)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 1000..1022
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(1016..1207)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
protein_bind 1425..1446
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 1461..1491
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1499..1515
/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 1554..2261
/codon_start=1
/gene="cI"
/product="phage lambda repressor"
/label=-lambda repressor
/translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
VGKVIASQWPEETF"
CDS 2265..2288
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 2589..2606
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS complement(2688..3479)
/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(3480..3551)
/gene="bla"
/label=AmpR promoter
CDS complement(3581..4663)
/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 4625..4644
/label=LacI-R
/note="LacI, reverse primer"
rep_origin 4746..588
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
61 gcggtggttt gtttgccgga tcaagagcta ccacctcttt ttccgaaggt aactggcttc
121 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
181 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
241 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
301 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
361 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
421 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
481 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
541 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
601 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
661 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
721 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
781 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
841 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
901 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
961 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
1021 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
1081 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
1141 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
1201 tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
1261 aacgagagag gatgctcacg atacgggtta ctgatgatga acaagaggac atccggtcaa
1321 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttgctgagga gacttaggga
1381 ccctggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
1441 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
1501 gtgagcggat aacaatttca cacacctgca ggtgcagtaa ggaggaaaaa aaaatgagca
1561 caaaaaagaa accattaaca caagagcagc ttgaggacgc acgtcgcctt aaagcaattt
1621 atgaaaaaaa gaaaaatgaa cttggcttat cccaggaatc tgtcgcagac aagatgggga
1681 tggggcagtc aggcgttggt gctttattta atggcatcaa tgcattaaat gcttataacg
1741 ccgcattgct tgcaaaaatt ctcaaagtta gcgttgaaga atttagccct tcaatcgcca
1801 gagaaatcta cgagatgtat gaagcggtta gtatgcagcc gtcacttaga agtgagtatg
1861 agtaccctgt tttttctcat gttcaggcag ggatgttctc acctgagctt agaaccttta
1921 ccaaaggtga tgcggagaga tgggtaagca caaccaaaaa agccagtgat tctgcattct
1981 ggcttgaggt tgaaggtaat tccatgaccg caccaacagg ctccaagcca agctttcctg
2041 acggaatgtt aattctcgtt gaccctgagc aggctgttga gccaggtgat ttctgcatag
2101 ccagacttgg gggtgatgag tttaccttca agaaactgat cagggatagc ggtcaggtgt
2161 ttttacaacc actaaaccca cagtacccaa tgatcccatg caatgagagt tgttccgttg
2221 tggggaaagt tatcgctagt cagtggcctg aagagacgtt tggggactac aaggacgacg
2281 atgacaagag tctggaaaaa cacagctggt atcatggccc tgtgagccgt aacgcggccg
2341 aatacctgct gagctctggc attaatggtt cttttctggt tcgtgaaagt gaaagtagcc
2401 cgggccagcg cagcatttct ctgcgttatg aaggtcgcgt gtatcactac cgtatcaaca
2461 ccgccagcga tggcaaactg tacgtttcta gtgaatctcg cttcaatacc ctggcagaac
2521 tggtgcatca ccatagcacg gttgcggatg gtctgatcac cacgctgcat tatccggcgc
2581 cgaaacgcca ccatcatcac catcattaac ttaattaacg gcactcctca gcaaatataa
2641 tgaccctctt gataacccaa gagggcattt tttaatgccc atggcgttta tttgccgact
2701 accttggtga tctcgccttt cacgtagtgg acaaattctt ccaactgatc tgcgcgcgag
2761 gccaagcgat cttcttcttg tccaagataa gcctgtctag cttcaagtat gacgggctga
2821 tactgggccg gcaggcgctc cattgcccag tcggcagcga catccttcgg cgcgattttg
2881 ccggttactg cgctgtacca aatgcgggac aacgtaagca ctacatttcg ctcatcgcca
2941 gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat ttagcgcctc aaatagatcc
3001 tgttcaggaa ccggatcaaa gagttcctcc gccgctggac ctaccaaggc aacgctatgt
3061 tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga tcgtggctgg ctcgaagata
3121 cctgcaagaa tgtcattgcg ctgccattct ccaaattgca gttcgcgctt agctggataa
3181 cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt ctacagcgcg gagaatctcg
3241 ctctctccag gggaagccga agtttccaaa aggtcgttga tcaaagctcg ccgcgttgtt
3301 tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat cactgtgtgg cttcaggccg
3361 ccatccactg cggagccgta caaatgtacg gccagcaacg tcggttcgag atggcgctcg
3421 atgacgccaa ctacctctga tagttgagtc gatacttcgg cgatcaccgc ttccctcata
3481 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
3541 atatttgaat gtatttagaa aaatattaat tgcgttgcgc tcactgcccg ctttccagtc
3601 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
3661 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc
3721 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
3781 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
3841 cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc
3901 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
3961 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
4021 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
4081 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
4141 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
4201 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
4261 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
4321 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
4381 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
4441 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
4501 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
4561 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
4621 caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt
4681 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcaccat aggccaaata
4741 ggccgt
//
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