pAIDA1 vector (V012813#)

Basic Vector Information

A construct with the AIDA surface expression system for display of a passenger protein flanked by His and Myc tags

      • Vector Name:
      • pAIDA1
      • Antibiotic Resistance:
      • Chloramphenicol
      • Length:
      • 5561 bp
      • Type:
      • E.coli expression plasmid
      • Source/Author:
      • Gen Larsson
      • Copy Number:
      • Low copy number
      • Promoter:
      • lacUV5
      • Cloning Method:
      • Enzyme Cut
      • 5' Primer:
      • GAGCGGATAACAATTTCACACAGG
      • Growth Strain(s):
      • DH5alpha
      • Growth Temperature:
      • 37℃

pAIDA1 vector Vector Map

pAIDA15561 bp60012001800240030003600420048005400lac UV5 promoterlac operatorSP6xHisHRV3CTEV siteMycADIAp15A oriL4440pENTR-RCmRlacI promoterlacI

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

Copy Sequence

Download GeneBank File(.gb)

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"

This page is informational only.