LOCUS       sequence-429713-        5226 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression vector for MBP-tagged mouse Dvl2 DEP domain 
            (residues 416-510).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5226)
  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 5226)
  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-mDvl2DEP(416-510)_pCDFduet
FEATURES             Location/Qualifiers
     source          1..5226
                     /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        1676..1694
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1695..1719
                     /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             1828..1872
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1910..1928)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1924..1971
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2142..2933)
                     /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(2934..3025)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3073..3811)
                     /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(4021..5103)
                     /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     5065..5084
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5104..5181)
                     /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 agggatccgg tctctctgtc
     1321 cacatggaca tggcctctgt caccaaggcc atggcagccc cagagtctgg gctcgaagtc
     1381 cgggaccgca tgtggctcaa gatcaccatc ccaaacgcct ttctaggctc cgatgtggtg
     1441 gactggctgt accatcacgt ggaaggtttt cctgagcgcc gggaggcccg caagtatgcc
     1501 agtgggctgt tgaaggcggg actcatccgg cacaccgtca acaagattac tttctctgag
     1561 cagtgctatt atgtcttcgg ggacctcagt ggttaatagt cgacaagctt gcggccgcat
     1621 aatgcttaag tcgaacagaa agtaatcgta ttgtacacgg ccgcataatc gaaattaata
     1681 cgactcacta taggggaatt gtgagcggat aacaattccc catcttagta tattagttaa
     1741 gtataagaag gagatataca tatggcagat ctcaattgga tatcggccgg ccacgcgatc
     1801 gctgacgtcg gtaccctcga gtctggtaaa gaaaccgctg ctgcgaaatt tgaacgccag
     1861 cacatggact cgtctactag cgcagcttaa ttaacctagg ctgctgccac cgctgagcaa
     1921 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaacct
     1981 caggcatttg agaagcacac ggtcacactg cttccggtag tcaataaacc ggtaaaccag
     2041 caatagacat aagcggctat ttaacgaccc tgccctgaac cgacgaccgg gtcatcgtgg
     2101 ccggatcttg cggcccctcg gcttgaacga attgttagac attatttgcc gactaccttg
     2161 gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctgcgcg cgaggccaag
     2221 cgatcttctt cttgtccaag ataagcctgt ctagcttcaa gtatgacggg ctgatactgg
     2281 gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat tttgccggtt
     2341 actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc gccagcccag
     2401 tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag atcctgttca
     2461 ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct atgttctctt
     2521 gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa gatacctgca
     2581 agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg ataacgccac
     2641 ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat ctcgctctct
     2701 ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca
     2761 agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag gccgccatcc
     2821 actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg ctcgatgacg
     2881 ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttccct catactcttc
     2941 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3001 gaatgtattt agaaaaataa acaaatagct agctcactcg gtcgctacgc tccgggcgtg
     3061 agactgcggc gggcgctgcg gacacataca aagttaccca cagattccgt ggataagcag
     3121 gggactaaca tgtgaggcaa aacagcaggg ccgcgccggt ggcgtttttc cataggctcc
     3181 gccctcctgc cagagttcac ataaacagac gcttttccgg tgcatctgtg ggagccgtga
     3241 ggctcaacca tgaatctgac agtacgggcg aaacccgaca ggacttaaag atccccaccg
     3301 tttccggcgg gtcgctccct cttgcgctct cctgttccga ccctgccgtt taccggatac
     3361 ctgttccgcc tttctccctt acgggaagtg tggcgctttc tcatagctca cacactggta
     3421 tctcggctcg gtgtaggtcg ttcgctccaa gctgggctgt aagcaagaac tccccgttca
     3481 gcccgactgc tgcgccttat ccggtaactg ttcacttgag tccaacccgg aaaagcacgg
     3541 taaaacgcca ctggcagcag ccattggtaa ctgggagttc gcagaggatt tgtttagcta
     3601 aacacgcggt tgctcttgaa gtgtgcgcca aagtccggct acactggaag gacagatttg
     3661 gttgctgtgc tctgcgaaag ccagttacca cggttaagca gttccccaac tgacttaacc
     3721 ttcgatcaaa ccacctcccc aggtggtttt ttcgtttaca gggcaaaaga ttacgcgcag
     3781 aaaaaaagga tctcaagaag atcctttgat cttttctact gaaccgctct agatttcagt
     3841 gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt
     3901 ctcatgttag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     3961 ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     4021 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4081 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     4141 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     4201 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     4261 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag
     4321 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     4381 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     4441 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     4501 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     4561 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
     4621 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
     4681 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     4741 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     4801 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     4861 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     4921 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     4981 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     5041 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     5101 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     5161 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
     5221 ggaaat
//
