LOCUS       pAIDA1.        5561 bp 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
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Microb Cell
            Fact."; date: "2012-09-3"; volume: "11:118. doi"; pages: " 
            10.1186/1475-2859-11-118"
FEATURES             Location/Qualifiers
     source          1..5561
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..31
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    39..55
                     /label=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)."
     CDS             77..229
                     /codon_start=1
                     /label=SP
                     /note="SP"
                     /translation="MNKAYSIIWSHSRQAWIVASELARGHGFVLAKNTLLVLAVVSTIG
                     NAFAVD"
     CDS             230..247
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             248..271
                     /codon_start=1
                     /label=HRV3C
                     /note="HRV3C"
                     /translation="LEALFQGP"
     CDS             299..319
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     CDS             320..349
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     CDS             356..1849
                     /codon_start=1
                     /label=ADIA
                     /note="ADIA"
                     /translation="VNNNGSIVINNSIINGNITNDADLSFGTAKLLSATVNGSLVNNKN
                     IILNPTKESAAAIGNTLTVSNYTGTPGSVISLGGVLEGDNSLTDRLVVKGNTSGQSDIV
                     YVNEDGSGGQTRDGINIISVEGNSDAEFSLKNRVVAGAYDYTLQKGNESGTDNKGWYLT
                     SHLPTSDTRQYRPENGSYATNMALANSLFLMDLNERKQFRAMSDNTQPESASVWMKITG
                     GISSGKLNDGQNKTTTNQFINQLGGDIYKFHAEQLGDFTLGIMGGYANAKGKTINYTSN
                     KAARNTLDGYSVGVYGTWYQNGENATGLFAETWMQYNWFNASVKGDGLEEEKYNLNGLT
                     ASAGGGYNLNVHTWTSPEGITGEFWLQPHLQAVWMGVTPDTHQEDNGTVVQGAGKNNIQ
                     TKAGIRASWKVKSTLDKDTGRRFRPYIEANWIHNTHEFGVKMSDDSQLLSGSRNQGEIK
                     TGIEGVITQNLSVNGGVAYQAGGHGSNAISGALGIKYSF"
     rep_origin      2344..2889
                     /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     3006..3023
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3473..3492)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3659..4315
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        4331..4408
                     /label=lacI promoter
     CDS             4409..5488
                     /label=lacI
                     /note="lac repressor"
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
//
