LOCUS       sequence-259529-        7287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human beta-catenin R1C (138–781) in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7287)
  AUTHORS   Xing Y, Takemaru K, Liu J, Berndt JD, Zheng JJ, Moon RT, Xu W
  TITLE     Crystal structure of a full-length beta-catenin.
  JOURNAL   Structure. 2008 Mar;16(3):478-87. doi: 10.1016/j.str.2007.12.021.
  PUBMED    18334222
REFERENCE   2  (bases 1 to 7287)
  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.str.2007.12"; journalName: "Structure"; date: "2008-03"; 
            volume: "16"; issue: "3"; pages: "478-87"
COMMENT     Sequence Label: pET28a-human Beta-catenin R1C  (138–781)
FEATURES             Location/Qualifiers
     source          1..7287
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..18
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             22..54
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             64..84
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2026..2043
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2096..2114)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2110..2157
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2194..2649
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2281..2300)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2491..2512
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2741..3556)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2741..3556)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3464..3483
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3548..3567
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3678..4266
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4167..4186
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4420..4437
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4452..4591
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4517..4536)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4677..4699
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4693..4884)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5693..6775)
                     /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     6737..6756
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6776..6853)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7059..7078
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     7162..7181
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7162..7180
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    7181..7205
                     /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)."
     RBS             7220..7242
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             7261..7278
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 ctggtgccgc gcggcagcca tatggctagc atgactggtg gacagcaaat gggtcgcgga
       61 tccgaaaacc tgtattttca gggcaacttg attaactatc aagatgatgc agaacttgcc
      121 acacgtgcaa tccctgaact gacaaaactg ctaaatgacg aggaccaggt ggtggttaat
      181 aaggctgcag ttatggtcca tcagctttct aaaaaggaag cttccagaca cgctatcatg
      241 cgttctcctc agatggtgtc tgctattgta cgtaccatgc agaatacaaa tgatgtagaa
      301 acagctcgtt gtaccgctgg gaccttgcat aacctttccc atcatcgtga gggcttactg
      361 gccatcttta agtctggagg cattcctgcc ctggtgaaaa tgcttggttc accagtggat
      421 tctgtgttgt tttatgccat tacaactctc cacaaccttt tattacacca agaaggagct
      481 aaaatggcag tgcgtttagc tggtgggctg cagaaaatgg ttgccttgct caacaaaaca
      541 aatgttaaat tcttggctat tacgacagac tgccttcaaa ttttagctta tggcaaccaa
      601 gaaagcaagc tcatcatact ggctagtggt ggaccccaag ctttagtaaa tataatgagg
      661 acctatactt acgaaaaact actgtggacc acaagcagag tgctgaaggt gctatctgtc
      721 tgctctagta ataagccggc tattgtagaa gctggtggaa tgcaagcttt aggacttcac
      781 ctgacagatc caagtcaacg tcttgttcag aactgtcttt ggactctcag gaatctttca
      841 gatgctgcaa ctaaacagga agggatggaa ggtctccttg ggactcttgt tcagcttctg
      901 ggttcagatg atataaatgt ggtcacctgt gcagctggaa ttctttctaa cctcacttgc
      961 aataattata agaacaagat gatggtctgc caagtgggtg gtatagaggc tcttgtgcgt
     1021 actgtccttc gggctggtga cagggaagac atcactgagc ctgccatctg tgctcttcgt
     1081 catctgacca gccgacacca agaagcagag atggcccaga atgcagttcg ccttcactat
     1141 ggactaccag ttgtggttaa gctcttacac ccaccatccc actggcctct gataaaggct
     1201 actgttggat tgattcgaaa tcttgccctt tgtcccgcaa atcatgcacc tttgcgtgag
     1261 cagggtgcca ttccacgact agttcagttg cttgttcgtg cacatcagga tacccagcgc
     1321 cgtacgtcca tgggtgggac acagcagcaa tttgtggagg gggtccgcat ggaagaaata
     1381 gttgaaggtt gtaccggagc ccttcacatc ctagctcggg atgttcacaa ccgaattgtt
     1441 atcagaggac taaataccat tccattgttt gtgcagctgc tttattctcc cattgaaaac
     1501 atccaaagag tagctgcagg ggtcctctgt gaacttgctc aggacaagga agctgcagaa
     1561 gctattgaag ctgagggagc cacagctcct ctgacagagt tacttcactc taggaatgaa
     1621 ggtgtggcga catatgcagc tgctgttttg ttccgaatgt ctgaggacaa gccacaagat
     1681 tacaagaaac ggctttcagt tgagctgacc agctctctct tcagaacaga gccaatggct
     1741 tggaatgaga ctgctgatct tggacttgat attggtgccc agggagaacc ccttggatat
     1801 cgccaggatg atcctagcta tcgttctttt cactctggtg gatatggcca ggatgccttg
     1861 ggtatggacc ccatgatgga acatgagatg ggtggccacc accctggtgc tgactatcca
     1921 gttgatgggc tgccagatct ggggcatgcc caggacctca tggatgggct gcctccaggt
     1981 gacagcaatc agctggcctg gtttgatact gacctgtaac tcgagcacca ccaccaccac
     2041 cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     2101 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     2161 aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     2221 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2281 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2341 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2401 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2461 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2521 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     2581 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     2641 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     2701 ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac tcatcgagca
     2761 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     2821 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     2881 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     2941 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     3001 aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     3061 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata
     3121 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     3181 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     3241 ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     3301 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     3361 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     3421 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     3481 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa gacgtttccc
     3541 gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg
     3601 ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3661 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3721 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3781 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     3841 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3901 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3961 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4021 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4081 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4141 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4201 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4261 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4321 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     4381 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     4441 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     4501 aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc
     4561 gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc
     4621 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     4681 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta
     4741 aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag
     4801 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag
     4861 ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg
     4921 ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca
     4981 tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca
     5041 gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca
     5101 gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt
     5161 ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt
     5221 cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg
     5281 ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat
     5341 gcgcacccgt ggggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt
     5401 ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga
     5461 caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc
     5521 tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat
     5581 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     5641 tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc
     5701 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5761 gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca
     5821 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
     5881 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
     5941 tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact
     6001 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
     6061 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
     6121 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag
     6181 ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt
     6241 gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa
     6301 tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg
     6361 cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga
     6421 cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta
     6481 ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa
     6541 tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt
     6601 tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt
     6661 ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg
     6721 tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca
     6781 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
     6841 attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag
     6901 cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag
     6961 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     7021 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     7081 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc
     7141 gagatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca
     7201 attcccctct agaaataatt ttgtttaact ttaagaagga gatataccat gggcagcagc
     7261 catcatcatc atcatcacag cagcggc
//
