Basic Vector Information
A construct with the AIDA surface expression system for display of a passenger protein flanked by His and Myc tags
pAIDA1 vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pAIDA1 vector Sequence
LOCUS Exported 5561 bp ds-DNA circular SYN 11-MAY-2023 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 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="lac repressor encoded by lacI lac operator" /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 77..229 /codon_start=1 /note="SP" /translation="MNKAYSIIWSHSRQAWIVASELARGHGFVLAKNTLLVLAVVSTIG NAFAVD" CDS 230..247 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /note="6xHis" /translation="HHHHHH" CDS 248..271 /codon_start=1 /note="HRV3C" /translation="LEALFQGP" CDS 299..319 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /note="TEV site" /translation="ENLYFQG" CDS 320..349 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /note="Myc" /translation="EQKLISEEDL" CDS 356..1849 /codon_start=1 /note="ADIA" /translation="VNNNGSIVINNSIINGNITNDADLSFGTAKLLSATVNGSLVNNKN IILNPTKESAAAIGNTLTVSNYTGTPGSVISLGGVLEGDNSLTDRLVVKGNTSGQSDIV YVNEDGSGGQTRDGINIISVEGNSDAEFSLKNRVVAGAYDYTLQKGNESGTDNKGWYLT SHLPTSDTRQYRPENGSYATNMALANSLFLMDLNERKQFRAMSDNTQPESASVWMKITG GISSGKLNDGQNKTTTNQFINQLGGDIYKFHAEQLGDFTLGIMGGYANAKGKTINYTSN KAARNTLDGYSVGVYGTWYQNGENATGLFAETWMQYNWFNASVKGDGLEEEKYNLNGLT ASAGGGYNLNVHTWTSPEGITGEFWLQPHLQAVWMGVTPDTHQEDNGTVVQGAGKNNIQ TKAGIRASWKVKSTLDKDTGRRFRPYIEANWIHNTHEFGVKMSDDSQLLSGSRNQGEIK TGIEGVITQNLSVNGGVAYQAGGHGSNAISGALGIKYSF" rep_origin 2344..2889 /direction=RIGHT /label=p15A ori /note="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 /note="lacI promoter" CDS 4409..5485 /codon_start=1 /gene="lacI" /product="lac repressor" /label=lacI /note="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"
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