| 质粒名称: | pAIDA1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 79180 |
| 存储实验室: | Gen Larsson |
|
载体骨架基本信息 |
|
| 载体名称: | pKM1D |
| 空载体大小: | 3800 bp |
| 完整质粒大小: | 3600 bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | 氯霉素25 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Adhesin Involved in Diffuse Adherence,AIDA |
| 物种来源: | Synthetic; E. coli |
| 插入大小: | 1800 bp |
| 启动子: | lacUV5 |
| 标签/融合蛋白: |
His（插入件上的N端子）
Myc（插入件上的C端子）
|
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | HindIII(未破坏) |
| 3’克隆位点: | BclI（克隆过程中销毁） |
| 5’测序引物 | GAGCGGATAACAATTTCACACAGG |
|
文献引用方法 |
|
| 材料和方法部分: | pAIDA1 was a gift from Gen Larsson (Addgene plasmid # 79180 ; http://n2t.net/addgene:79180 ; RRID:Addgene_79180) |
| 参考文献部分: | A dual tag system for facilitated detection of surface expressed proteins in Escherichia coli. Jarmander J, Gustavsson M, Do TH, Samuelson P, Larsson G. Microb Cell Fact. 2012 Sep 3;11:118. doi: 10.1186/1475-2859-11-118. 10.1186/1475-2859-11-118 PubMed 22943700 |
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 pAIDA1 5561 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION A construct with the AIDA surface expression system for display of a
passenger protein flanked by His and Myc tags.
ACCESSION .
VERSION .
KEYWORDS pAIDA1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5561)
AUTHORS Jarmander J, Gustavsson M, Do TH, Samuelson P, Larsson G
TITLE A dual tag system for facilitated detection of surface expressed
proteins in Escherichia coli.
JOURNAL Microb Cell Fact. 2012 Sep 3;11:118. doi: 10.1186/1475-2859-11-118.
PUBMED 22943700
REFERENCE 2 (bases 1 to 5561)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5561
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..31
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
protein_bind 39..55
/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)."
primer_bind 44..66
/label=M13/pUC Reverse
/note="In lacZ gene"
CDS 230..247
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 299..319
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
CDS 320..349
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
rep_origin 2344..2889
/direction=RIGHT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin.
"
primer_bind 2790..2809
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3006..3023
/label=L4440
/note="L4440 vector, forward primer"
primer_bind complement(3473..3492)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 3659..4318
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(3725..3744)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter 4331..4408
/gene="lacI"
/label=lacI promoter
CDS 4409..5485
/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
ALADSLMQLARQVSRLESG"
primer_bind complement(4428..4447)
/label=LacI-R
/note="LacI, reverse primer"
ORIGIN
1 tttacacttt atgcttccgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
61 cacaggaaag cttcatatga ataaggccta cagtatcatt tggagccact ccagacaggc
121 ctggattgtg gcctcagagt tagccagagg acatggtttt gtccttgcaa aaaatacact
181 gctggtattg gcggttgttt ccacaatcgg aaatgcattt gcagtcgacc accatcacca
241 tcaccatctg gaagcgctgt tccagggtcc gggtacccag aaacagcgta ccgagctcga
301 aaacctgtac ttccagggtg aacagaaact gattagcgaa gaagatctgt ctagagtgaa
361 taacaatgga agcattgtca ttaataacag cattataaac gggaatatta cgaatgatgc
421 tgacttaagt tttggtacag caaagctgct ctctgctaca gtgaatggta gtcttgttaa
481 taacaaaaat atcattctta atcctacaaa agaaagtgcg gccgctatag gtaatactct
541 taccgtgtca aattatactg ggacaccggg aagtgttatt tctcttggtg gtgtgcttga
601 aggagataat tcacttacgg accgtctggt ggtgaaaggt aatacctctg gtcaaagtga
661 catcgtttat gtcaatgaag atggcagtgg tggtcagacg agagatggta ttaatattat
721 ttctgtagag ggaaattctg atgcagaatt ctctctgaag aaccgcgtag ttgccggagc
781 ttatgattac acactgcaga aaggaaacga gagtgggaca gataataagg gatggtattt
841 aaccagtcat cttcccacat ctgatacccg gcaatacaga ccggagaacg gaagttatgc
901 taccaatatg gcactggcta actcactgtt cctcatggat ttgaatgagc gtaagcaatt
961 cagggccatg agtgataata cacagcctga gtctgcatcc gtgtggatga agatcactgg
1021 aggaataagc tctggtaagc tgaatgacgg gcaaaataaa acaacaacca atcagtttat
1081 caatcagctc gggggggata tttataaatt ccatgctgaa caactgggtg attttacctt
1141 agggattatg ggaggatacg cgaatgcaaa aggtaaaacg ataaattaca cgagcaacaa
1201 agctgccaga aacacactgg atggttattc tgtcggggta tacggtacgt ggtatcagaa
1261 tggggaaaat gcaacagggc tctttgctga aacttggatg caatataact ggtttaatgc
1321 atcagtgaaa ggtgacggac tggaagaaga aaaatataat ctgaatggtt taaccgcttc
1381 tgcaggtggg ggatataacc tgaatgtgca cacatggaca tcacctgaag gaataacagg
1441 tgaattctgg ttacagcctc atttgcaggc tgtctggatg ggggttacac cggatacaca
1501 tcaggaggat aacggaacgg tggtgcaggg agcagggaaa aataatattc agacaaaagc
1561 aggtattcgt gcatcctgga aggtgaaaag caccctggat aaggataccg ggcggaggtt
1621 ccgtccgtat atagaggcaa actggatcca taacactcat gaatttggtg ttaaaatgag
1681 tgatgacagc cagttgttgt caggtagccg aaatcaggga gagataaaga caggtattga
1741 aggggtgatt actcaaaact tgtcagtgaa tggcggagtc gcatatcagg caggaggtca
1801 cgggagcaat gccatctccg gagcactggg gataaaatac agcttctgat aatgatcctg
1861 gcacgcggcg cgccccttgg tgcgcaaact attaactggc gaactactta ctctagcttc
1921 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
1981 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
2041 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
2101 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
2161 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
2221 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
2281 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccttaa taagatgatc
2341 ttcttgagat cgttttggtc tgcgcgtaat ctcttgctct gaaaacgaaa aaaccgcctt
2401 gcagggcggt ttttcgaagg ttctctgagc taccaactct ttgaaccgag gtaactggct
2461 tggaggagcg cagtcaccaa aacttgtcct ttcagtttag ccttaaccgg cgcatgactt
2521 caagactaac tcctctaaat caattaccag tggctgctgc cagtggtgct tttgcatgtc
2581 tttccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg gactgaacgg
2641 ggggttcgtg catacagtcc agcttggagc gaactgccta cccggaactg agtgtcaggc
2701 gtggaatgag acaaacgcgg ccataacagc ggaatgacac cggtaaaccg aaaggcagga
2761 acaggagagc gcacgaggga gccgccaggg ggaaacgcct ggtatcttta tagtcctgtc
2821 gggtttcgcc accactgatt tgagcgtcag atttcgtgat gcttgtcagg ggggcggagc
2881 ctatggaaaa acggctttgc cgcggccctc tcacttccct gttaagtatc ttcctggcat
2941 cttccaggaa atctccgccc cgttcgtaag ccatttccgc tcgccgcagt cgaacgaccg
3001 agcgtagcga gtcagtgagc gaggaagcgg aatatatcct gtatcacata ttctgctgac
3061 gcaccggtgc agcctttttt ctcctgccac atgaagcact tcactgacac cctcatcagt
3121 gccaacatag taagccagta tacactccgc tagcgctgag gtctgcctcg tgaagaaggt
3181 gttgctgact cataccaggc ctgaatcgcc ccatcatcca gccagaaagt gagggagcca
3241 cggttgatga gagctttgtt gtaggtggac cagttggtga ttttgaactt ttgctttgcc
3301 acggaacggt ctgcgttgtc gggaagatgc gtgatctgat ccttcaactc agcaaaagtt
3361 cgatttattc aacaaagcca cgttgtgtct caaaatctct gatgttacat tgcacaagat
3421 aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
3481 gttatgagcc atattcaacg ggaaacgtct tgctcgagta tccgctcatg agattatcaa
3541 aaaggatctt cacctagatc cttttgtaag aggttccaac tttcaccata atgaaataag
3601 atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg aagctaaaat
3661 ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca
3721 ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat
3781 tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca
3841 cattcttgcc cgcctgatga atgctcatcc ggagtttcgt atggcaatga aagacggtga
3901 gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac
3961 gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc
4021 gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa
4081 tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc
4141 caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga
4201 caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt
4261 cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaatt
4321 tttttaaggc gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc
4381 ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga
4441 gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc
4501 tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg
4561 cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct
4621 ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg
4681 tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt
4741 gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca
4801 ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc
4861 tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt
4921 ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc
4981 tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca
5041 gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca
5101 aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct
5161 gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt
5221 gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca
5281 ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca
5341 ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc
5401 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
5461 acaggtttcc cgactggaaa gcgggcaagt gagtggataa ccgtattacc gcctttgagt
5521 gagctgatac cgggaattct cactcattag gcaccccagg c
//
