LOCUS       sequence-308818-        7120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MBP-Axin_384-518-His (human Axin minimal scaffold fragment
            as a fusion protein with MBP and His- tags) in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7120)
  AUTHORS   Gavagan M, Fagnan E, Speltz EB, Zalatan JG
  TITLE     The Scaffold Protein Axin Promotes Signaling Specificity within the 
            Wnt Pathway by Suppressing Competing Kinase Reactions.
  JOURNAL   Cell Syst. 2020 Jun 24;10(6):515-525.e5. doi: 
            10.1016/j.cels.2020.05.002. Epub 2020 Jun 17.
  PUBMED    32553184
REFERENCE   2  (bases 1 to 7120)
  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"; doi: 
            "10.1016/j.cels.2020.05.002"; journalName: "Cell Syst"; date: 
            "2020-06-24- 24"; volume: "10"; issue: "6"; pages: "515-525.e5"
COMMENT     Sequence Label: pMG023 MBP-Axin 384-518-His (miniAxin)
FEATURES             Location/Qualifiers
     source          1..7120
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..78
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             79..1161
                     /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     complement(98..117)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1404..1409,1410..1425,1426..1432)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1440..1456
                     /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)."
     primer_bind     1445..1467
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1526..2626
                     /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"
     regulatory      2483..2492
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2600..2623
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             2675..2686
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2729..2749
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3167..3184
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3211..3228)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3211..3227)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3220..3242)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3406..3423)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3576..3662
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3754..3781
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3800..3891
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3892..3960,3961..4752)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4110..4129)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4794..5307)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4989..5010)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5201..5220
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      5418..6006
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5907..5926
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6160..6177
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6192..6332
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6258..6277)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6418..6440
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6434..6625)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gacaccatcg aatggtgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt
       61 caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt
      121 gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg
      181 cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa
      241 caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac
      301 gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg
      361 gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt
      421 ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt
      481 gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca
      541 cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg
      601 gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg
      661 cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg
      721 gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat
      781 gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg
      841 cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac
      901 gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc
      961 ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag
     1021 ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg
     1081 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     1141 cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc
     1201 acaattctca tgtttgacag cttatcatcg actgcacggt gcaccaatgc ttctggcgtc
     1261 aggcagccat cggaagctgt ggtatggctg tgcaggtcgt aaatcactgc ataattcgtg
     1321 tcgctcaagg cgcactcccg ttctggataa tgttttttgc gccgacatca taacggttct
     1381 ggcaaatatt ctgaaatgag ctgttgacaa ttaatcatcg gctcgtataa tgtgtggaat
     1441 tgtgagcgga taacaatttc acacaggaaa cagccagtcc gtttaggtgt tttcacgagc
     1501 acttcaccaa caaggaccat agattatgaa aactgaagaa ggtaaactgg taatctggat
     1561 taacggcgat aaaggctata acggtctcgc tgaagtcggt aagaaattcg agaaagatac
     1621 cggaattaaa gtcaccgttg agcatccgga taaactggaa gagaaattcc cacaggttgc
     1681 ggcaactggc gatggccctg acattatctt ctgggcacac gaccgctttg gtggctacgc
     1741 tcaatctggc ctgttggctg aaatcacccc ggacaaagcg ttccaggaca agctgtatcc
     1801 gtttacctgg gatgccgtac gttacaacgg caagctgatt gcttacccga tcgctgttga
     1861 agcgttatcg ctgatttata acaaagatct gctgccgaac ccgccaaaaa cctgggaaga
     1921 gatcccggcg ctggataaag aactgaaagc gaaaggtaag agcgcgctga tgttcaacct
     1981 gcaagaaccg tacttcacct ggccgctgat tgctgctgac gggggttatg cgttcaagta
     2041 tgaaaacggc aagtacgaca ttaaagacgt gggcgtggat aacgctggcg cgaaagcggg
     2101 tctgaccttc ctggttgacc tgattaaaaa caaacacatg aatgcagaca ccgattactc
     2161 catcgcagaa gctgccttta ataaaggcga aacagcgatg accatcaacg gcccgtgggc
     2221 atggtccaac atcgacacca gcaaagtgaa ttatggtgta acggtactgc cgaccttcaa
     2281 gggtcaacca tccaaaccgt tcgttggcgt gctgagcgca ggtattaacg ccgccagtcc
     2341 gaacaaagag ctggcaaaag agttcctcga aaactatctg ctgactgatg aaggtctgga
     2401 agcggttaat aaagacaaac cgctgggtgc cgtagcgctg aagtcttacg aggaagagtt
     2461 ggcgaaagat ccacgtattg ccgccaccat ggaaaacgcc cagaaaggtg aaatcatgcc
     2521 gaacatcccg cagatgtccg ctttctggta tgccgtgcgt actgcggtga tcaacgccgc
     2581 cagcggtcgt cagactgtcg atgaagccct gaaagacgcg cagactaatt cgagctcgaa
     2641 caacaacaac aataacaata acaacaacct cgggatcgag ggaaggattt caactagtgg
     2701 atctggtggt ggtggcggtt ccatgagcga gaatctttat tttcagggat ccgagcctca
     2761 gaagttcgcg gaggagctca tccaccgcct ggaggctgtg cagcgcacgc gggaggccga
     2821 ggagaagctg gaggagcggc tgaagcgcgt gcgcatggag gaggaaggtg aggacggcga
     2881 tccatcgtca gggcccccag ggccgtgtca caagctgcct cccgcccccg cttggcacca
     2941 cttcccgccc cgctgtgtgg acatgggctg tgccgggctc cgggatgcac acgaggagaa
     3001 ccctgagagc atcctggacg agcacgtaca gcgtgtgctg aggacacctg gccgccagtc
     3061 gcctgggcct ggccatcgct ccccggacag tgggcacgtg gccaagatgc cagtggcact
     3121 ggggggtgcc gcctcggggc acgggaagca cgtacccagc ggccgccacc atcaccacca
     3181 ccactagggg ccgcactcga gaagcttggc actggccgtc gttttacaac gtcgtgactg
     3241 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     3301 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     3361 cgaatggcag cttggctgtt ttggcggatg agataagatt ttcagcctga tacagattaa
     3421 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt
     3481 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
     3541 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
     3601 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     3661 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
     3721 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
     3781 ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3841 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3901 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3961 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4021 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     4081 ttcccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     4141 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     4201 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4261 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     4321 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4381 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     4441 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4501 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4561 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4621 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4681 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4741 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaccccgg
     4801 ttgataatca gaaaagcccc aaaaacagga agattgtata agcaaatatt taaattgtaa
     4861 acgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     4921 aataggccga aatcggcaaa atcccttata aatcaaaaga atagcccgag atagggttga
     4981 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     5041 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc aaatcaagtt
     5101 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     5161 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     5221 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     5281 cgcttaatgc gccgctacag ggcgcgtaaa aggatctagg tgaagatcct ttttgataat
     5341 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5401 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5461 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5521 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     5581 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5641 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5701 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5761 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5821 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5881 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5941 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6001 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6061 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     6121 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     6181 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     6241 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
     6301 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     6361 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     6421 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     6481 aaagctcatc agcgtggtcg tgcagcgatt cacagatgtc tgcctgttca tccgcgtcca
     6541 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     6601 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta atggggattt ctgttcatgg
     6661 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
     6721 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc
     6781 agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac
     6841 agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact
     6901 tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg
     6961 tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct
     7021 gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca
     7081 tgcgcacccg tggccaggac ccaacgctgc ccgaaattcc
//
