LOCUS       sequence-429716-        5211 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression vector for MBP-tagged mouse Dvl2 PDZ domain 
            (residues 264-353).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5211)
  AUTHORS   Mahoney JP, Bruguera ES, Vasishtha M, Killingsworth LB, Kyaw S, Weis
            WI
  TITLE     PI(4,5)P(2)-stimulated positive feedback drives the recruitment of 
            Dishevelled to Frizzled in Wnt-beta-catenin signaling.
  JOURNAL   Sci Signal. 2022 Aug 23;15(748):eabo2820. doi: 
            10.1126/scisignal.abo2820. Epub 2022 Aug 23.
  PUBMED    35998232
REFERENCE   2  (bases 1 to 5211)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci 
            Signal."; date: "2022-08-23"; volume: "15(748):eabo2820. doi"; 
            pages: " 10.1126/scisignal.abo2820. Epub 2022 Aug 23"
COMMENT     Sequence Label: 6xHis-MBP-TEV-mDvl2PDZ(264-353)_pCDFduet
FEATURES             Location/Qualifiers
     source          1..5211
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     CDS             97..114
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..1227
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1201..1224
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1285..1305
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     promoter        1661..1679
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1680..1704
                     /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)."
     CDS             1813..1857
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1895..1913)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1909..1956
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2127..2918)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(2919..3010)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3058..3796)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(4006..5088)
                     /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
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5050..5069
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5089..5166)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taataaggag atataccatg ggcagccatc accatcatca ccacggaagt
      121 agcgatatga aaactgaaga aggtaaactg gtaatctgga ttaacggcga taaaggctat
      181 aacggtctcg ctgaagtcgg taagaaattc gagaaagata ccggaattaa agtcaccgtt
      241 gagcatccgg ataaactgga agagaaattc ccacaggttg cggcaactgg cgatggccct
      301 gacattatct tctgggcaca cgaccgcttt ggtggctacg ctcaatctgg cctgttggct
      361 gaaatcaccc cggacaaagc gttccaggac aagctgtatc cgtttacctg ggatgccgta
      421 cgttacaacg gcaagctgat tgcttacccg atcgctgttg aagcgttatc gctgatttat
      481 aacaaagatc tgctgccgaa cccgccaaaa acctgggaag agatcccggc gctggataaa
      541 gaactgaaag cgaaaggtaa gagcgcgctg atgttcaacc tgcaagaacc gtacttcacc
      601 tggccgctga ttgctgctga cgggggttat gcgttcaagt atgaaaacgg caagtacgac
      661 attaaagacg tgggcgtgga taacgctggc gcgaaagcgg gtctgacctt cctggttgac
      721 ctgattaaaa acaaacacat gaatgcagac accgattact ccatcgcaga agctgccttt
      781 aataaaggcg aaacagcgat gaccatcaac ggcccgtggg catggtccaa catcgacacc
      841 agcaaagtga attatggtgt aacggtactg ccgaccttca agggtcaacc atccaaaccg
      901 ttcgttggcg tgctgagcgc aggtattaac gccgccagtc cgaacaaaga gctggcaaaa
      961 gagttcctcg aaaactatct gctgactgat gaaggtctgg aagcggttaa taaagacaaa
     1021 ccgctgggtg ccgtagcgct gaagtcttac gaggaagagt tggcgaaaga tccacgtatt
     1081 gccgccacta tggaaaacgc ccagaaaggt gaaatcatgc cgaacatccc gcagatgtcc
     1141 gctttctggt atgccgtgcg tactgcggtg atcaacgccg ccagcggtcg tcagactgtc
     1201 gatgaagccc tgaaagacgc gcagactaat tcgagctcga acaacaacaa caataacaat
     1261 aacaacaacc tcgggccggg tgcggaaaac ctgtactttc agggatccaa catcatcacg
     1321 gtcacgctca acatggagaa gtacaacttc ctgggcatct ccattgtggg ccaaagtaac
     1381 gagcgtggtg atgggggcat ctacatcggc tccatcatga aagggggcgc ggtggctgcg
     1441 gacgggcgga tcgagcctgg agacatgctt ttgcaggtaa atgatatgaa ctttgagaac
     1501 atgagcaacg acgatgctgt acgagtgctg agagacatcg tgcacaagcc aggccccatc
     1561 gtgctcaccg tggccaagtg atagtcgaca agcttgcggc cgcataatgc ttaagtcgaa
     1621 cagaaagtaa tcgtattgta cacggccgca taatcgaaat taatacgact cactataggg
     1681 gaattgtgag cggataacaa ttccccatct tagtatatta gttaagtata agaaggagat
     1741 atacatatgg cagatctcaa ttggatatcg gccggccacg cgatcgctga cgtcggtacc
     1801 ctcgagtctg gtaaagaaac cgctgctgcg aaatttgaac gccagcacat ggactcgtct
     1861 actagcgcag cttaattaac ctaggctgct gccaccgctg agcaataact agcataaccc
     1921 cttggggcct ctaaacgggt cttgaggggt tttttgctga aacctcaggc atttgagaag
     1981 cacacggtca cactgcttcc ggtagtcaat aaaccggtaa accagcaata gacataagcg
     2041 gctatttaac gaccctgccc tgaaccgacg accgggtcat cgtggccgga tcttgcggcc
     2101 cctcggcttg aacgaattgt tagacattat ttgccgacta ccttggtgat ctcgcctttc
     2161 acgtagtgga caaattcttc caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt
     2221 ccaagataag cctgtctagc ttcaagtatg acgggctgat actgggccgg caggcgctcc
     2281 attgcccagt cggcagcgac atccttcggc gcgattttgc cggttactgc gctgtaccaa
     2341 atgcgggaca acgtaagcac tacatttcgc tcatcgccag cccagtcggg cggcgagttc
     2401 catagcgtta aggtttcatt tagcgcctca aatagatcct gttcaggaac cggatcaaag
     2461 agttcctccg ccgctggacc taccaaggca acgctatgtt ctcttgcttt tgtcagcaag
     2521 atagccagat caatgtcgat cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc
     2581 tgccattctc caaattgcag ttcgcgctta gctggataac gccacggaat gatgtcgtcg
     2641 tgcacaacaa tggtgacttc tacagcgcgg agaatctcgc tctctccagg ggaagccgaa
     2701 gtttccaaaa ggtcgttgat caaagctcgc cgcgttgttt catcaagcct tacggtcacc
     2761 gtaaccagca aatcaatatc actgtgtggc ttcaggccgc catccactgc ggagccgtac
     2821 aaatgtacgg ccagcaacgt cggttcgaga tggcgctcga tgacgccaac tacctctgat
     2881 agttgagtcg atacttcggc gatcaccgct tccctcatac tcttcctttt tcaatattat
     2941 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     3001 aataaacaaa tagctagctc actcggtcgc tacgctccgg gcgtgagact gcggcgggcg
     3061 ctgcggacac atacaaagtt acccacagat tccgtggata agcaggggac taacatgtga
     3121 ggcaaaacag cagggccgcg ccggtggcgt ttttccatag gctccgccct cctgccagag
     3181 ttcacataaa cagacgcttt tccggtgcat ctgtgggagc cgtgaggctc aaccatgaat
     3241 ctgacagtac gggcgaaacc cgacaggact taaagatccc caccgtttcc ggcgggtcgc
     3301 tccctcttgc gctctcctgt tccgaccctg ccgtttaccg gatacctgtt ccgcctttct
     3361 cccttacggg aagtgtggcg ctttctcata gctcacacac tggtatctcg gctcggtgta
     3421 ggtcgttcgc tccaagctgg gctgtaagca agaactcccc gttcagcccg actgctgcgc
     3481 cttatccggt aactgttcac ttgagtccaa cccggaaaag cacggtaaaa cgccactggc
     3541 agcagccatt ggtaactggg agttcgcaga ggatttgttt agctaaacac gcggttgctc
     3601 ttgaagtgtg cgccaaagtc cggctacact ggaaggacag atttggttgc tgtgctctgc
     3661 gaaagccagt taccacggtt aagcagttcc ccaactgact taaccttcga tcaaaccacc
     3721 tccccaggtg gttttttcgt ttacagggca aaagattacg cgcagaaaaa aaggatctca
     3781 agaagatcct ttgatctttt ctactgaacc gctctagatt tcagtgcaat ttatctcttc
     3841 aaatgtagca cctgaagtca gccccatacg atataagttg taattctcat gttagtcatg
     3901 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
     3961 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     4021 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4081 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     4141 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     4201 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     4261 ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat
     4321 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     4381 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     4441 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     4501 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     4561 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     4621 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     4681 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     4741 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     4801 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     4861 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     4921 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     4981 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     5041 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     5101 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     5161 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaaa t
//
