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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