| 质粒名称: | pAW240 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 170791 |
| 存储实验室: | Chang Liu |
|
载体骨架基本信息 |
|
| 载体名称: | ColE1 |
| 空载体大小: | 2244 bp |
| 完整质粒大小: | 2394 bp |
| 载体类型: | 酵母表达 |
| 筛选标记: | TRP1 |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
|
插入基因信息 |
|
| 插入基因名称: | p10B2-AT110 |
| 物种来源: | Synthetic |
| 启动子: | 10B2 (P1 specific promoter) |
| 克隆方法: | Gibson克隆 |
| 5’测序引物 | gttggccgattcattaatgcag |
| 3’测序引物 | AACCTCTGACACATGCAGCTCC |
|
文献引用方法 |
|
| 材料和方法部分: | pAW240 was a gift from Chang Liu (Addgene plasmid # 170791 ; http://n2t.net/addgene:170791 ; RRID:Addgene_170791) |
| 参考文献部分: | Rapid generation of potent antibodies by autonomous hypermutation in yeast. Wellner A, McMahon C, Gilman MSA, Clements JR, Clark S, Nguyen KM, Ho MH, Hu VJ, Shin JE, Feldman J, Hauser BM, Caradonna TM, Wingler LM, Schmidt AG, Marks DS, Abraham J, Kruse AC, Liu CC. Nat Chem Biol. 2021 Jun 24. pii: 10.1038/s41589-021-00832-4. doi: 10.1038/s41589-021-00832-4. 10.1038/s41589-021-00832-4 PubMed 34168368 |
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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 pAW240 4638 bp ds-DNA circular SYN 05-AUG-2021
DEFINITION P1 integration cassette for AT110 Polymerase for integration onto
Orthorep for Nanobody evolution.
ACCESSION .
VERSION .
KEYWORDS pAW240
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4638)
TITLE AT110 Nanobody P1 Integration Cassette
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 4638)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 5, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4638
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 118..137
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 371..388
/label=L4440
/note="L4440 vector, forward primer"
CDS 1335..1346
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
CDS 1347..1373
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
misc_RNA 1744..1795
/label=sTRSV HHRz
/note="hammerhead ribozyme from the tobacco ringspot virus
satellite RNA (Khvorova et al., 2003)"
terminator 1802..1967
/gene="S. cerevisiae ADH1"
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
primer_bind complement(1857..1881)
/label=pHybLex-R
/note="ADH terminator, reverse primer"
CDS complement(2039..2710)
/codon_start=1
/gene="S. cerevisiae TRP1"
/product="phosphoribosylanthranilate isomerase, required
for tryptophan biosynthesis"
/label=TRP1
/note="yeast auxotrophic marker"
/translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
KK"
primer_bind 2999..3021
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
promoter complement(3961..4052)
/gene="bla"
/label=AmpR promoter
terminator 4119..4166
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 4162..4180
/label=T7 Term
/note="T7 terminator, reverse primer"
rep_origin 4267..217
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
61 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
121 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
181 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
241 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
301 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
361 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
421 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
481 agcgggcagt gagcgcaacg caatagtact ataatatatg aattacatta ttaatttaaa
541 aacgcgtgat gacctataca taggaagatc tatagaaaca aaaagattaa taactttcaa
601 atatcagaaa aatatagaaa catgtgataa gctcatagac atataaagga tccatgagat
661 tcccatctat cttcaccgct gctttgttcg ctgcttcttc tgctttggct gctccagcta
721 acaccaccac cgaagacgaa accgctcaaa tcccagctga agctgttatc gactactctg
781 acttggaagg tgacttcgac gctgctgctt tgccattgtc taactctacc aacaacggtt
841 tgtcttctac caacaccacc atcgcttcta tcgctgctaa ggaagaaggt gttcaattgg
901 acaagagaga agctgacgca caggtgcact cctcgctgca ggaaagcggc ggcggcctgg
961 tgcaggcggg cggcagcctg cgcctgagct gcgcggcgag cggcaatatt tttgacgctg
1021 acatcatggg ctggtatcgc caggcgccgg gcaaagaacg cgaacttgtt gccagtatta
1081 ctgacggtgg tagtaccaat tatgcggata gcgtgaaagg ccgctttacc attagccgcg
1141 ataacgcgaa aaacaccgtg tatctgcaga tgaacagcct gaaaccggaa gataccgcgg
1201 tgtattattg cgcggctatc gcttacccgg acatcccgac ttacttcgac tacgactctg
1261 actatttcta ttggggccag ggcacccagg tgaccgtgag aggaggcagc ggatccaagg
1321 acaatagctc gacgattgaa ggtagatacc catacgacgt tccagactac gctctgcagg
1381 ctagtggtgg tggtggttct ggtggtggtg gttctggtgg tggtggttct gctagcgacg
1441 tccaggaact gacaactata tgcgagcaaa tcccctcacc aactttagaa tcgacgccgt
1501 actctttgtc aacgactact attttggcca acgggaaggc aatgcaagga gtttttgaat
1561 attacaaatc agtaacgttt gtcagtaatt gcggttctca cccctcaaca actagcaaag
1621 gcagccccat aaacacacag tatgtttttt gaagaagact tgtcatgaaa aaaaaaaaaa
1681 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
1741 aaacctgtca ccggatgtgt tttccggtct gatgagtccg tgaggacgaa acagggaatt
1801 cgcgaatttc ttatgattta tgatttttat tattaaataa gttataaaaa aaataagtgt
1861 atacaaattt taaagtgact cttaggtttt aaaacgaaaa ttcttattct tgagtaactc
1921 tttcctgtag gtcaggttgc tttctcaggt atagcatgag gtcgctcgag ctccgtttct
1981 attatgaatt tcatttataa agtttatgta caaatatcat aaaaaaagag aatctttttt
2041 tcttagcatt tttgacgaaa tttgctattt tgttagagtc ttttacacca tttgtctcca
2101 cacctccgct tacatcaaca ccaataacgc catttaatct aagcgcatca ccaacatttt
2161 ctggcgtcag tccaccagct aacataaaat gtaagctttc ggggctctct tgccttccaa
2221 cccagtcaga aatcgagttc caatccaaaa gttcacctgt cccacctgct tctgaatcaa
2281 acaagggaat aaacgaatga ggtttctgtg aagctgcact gagtagtatg ttgcagtctt
2341 ttggaaatac gagtctttta ataactggca aaccgaggaa ctcttggtat tcttgccacg
2401 actcatctcc atgcagttgg acgatatcaa tgccgtaatc attgaccaga gccaaaacat
2461 cctccttagg ttgattacga aacacgccaa ccaagtattt cggagtgcct gaactatttt
2521 tatatgcttt tacaagactt gaaattttcc ttgcaataac cgggtcaatt gttctctttc
2581 tattgggcac acatataata cccagcaagt cagcatcgga atctagagca cattctgcgg
2641 cctctgtgct ctgcaagccg caaactttca ccaatggacc agaactacct gtgaaattaa
2701 taacagacat tttataatta ttataaaaac tttcatataa gagatataaa atttaatatg
2761 gaagaaataa gacaagaaag atacaaccaa atgaaagaag ctctaaatag tgttgaaggt
2821 tataaaggaa aaattgtagc ctcagactca gattggtgtt tcaaagatcc tcaaggcaat
2881 agaataacag attttgatag tactttaagc cagccccgac acccgccaac acccgctgac
2941 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
3001 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaaggca
3061 aaaaaaaagg ctccaaaagg agcctttaat tgtatcggtt taccaatgct taatcagtga
3121 agcccctatt tcagcaatct gtctatttcg ttcgtccata gttgcctgac tacccgtcgt
3181 gtagataact acgatacggg aaggcttacc atctggtcca agtgcggcaa tgataccccg
3241 tgaaccacgc tctcctgccc cagatttatc agcaataaac cagccagcag gaagtgccga
3301 acgcagtagt ggtcctgcaa ctttatccgc ttcaagccag tctattaatt gttgccgtga
3361 agccagagta agtagttcgc cagttaatag tttgcgtaat gttgttgcca ttgccgcagg
3421 cattgtggtg tctcgctcgt cgtttggtat ggcttcattc agttctggtt cccaacggtc
3481 aaggcgagtt acatggtctc ccatattgtg caaaaaagcg gttaattcct tcggtccacc
3541 gatggttgtc agaagtaagt tcgccgcagt gttatcactc atcgttatgg cggcactaca
3601 taattctctt accgtcattc cgtccgtaag gtgcttttct gtcactgggg agtattcaac
3661 caagtcattc tgcgaatagt gtattcgtcg tccgagttgc tcttgccccg catctacacg
3721 ggataatact gctccacata gcagaacttt gaaagtgctc atcattggaa aacgctcttc
3781 agggcgaaaa ctctcaagga ttttaccgct attcaagtcc agttctatgt atcccactcg
3841 tgcccccaat tggtcctctg cgtcttttac tttcacgagc gtttctgggt gagcaaaaac
3901 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
3961 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctga tgagcggata
4021 catatttgaa tgtatttaga aaaataaaca aataggggaa ttaaaaaaaa gcccgctcat
4081 taggcgggct actagtcgga tatagttcct cctttcagca aaaaacccct caagacccgt
4141 ttagaggccc caaggggtta tgctagttat tgctcagcgg tggcagcagc caactcagct
4201 tcctttcggg ctttgttagc agccggatct tctagaagac cccgtagaaa agatcaaagg
4261 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
4321 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
4381 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
4441 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
4501 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
4561 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
4621 aacgacctac accgaact
//
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