LOCUS       sequence-259530-        6127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human APC-2,3 (1362–1540) in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6127)
  AUTHORS   Xing Y, Clements WK, Kimelman D, Xu W
  TITLE     Crystal structure of a beta-catenin/axin complex suggests a 
            mechanism for the beta-catenin destruction complex.
  JOURNAL   Genes Dev. 2003 Nov 15;17(22):2753-64. doi: 10.1101/gad.1142603. 
            Epub 2003 Nov 4.
  PUBMED    14600025
REFERENCE   2  (bases 1 to 6127)
  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: 
            "10.1101/gad.1142603"; journalName: "Genes Dev"; date: "2003-11-15- 
            15"; volume: "17"; issue: "22"; pages: "2753-64"
COMMENT     Sequence Label: pGEX-4T1 human APC-2,3 (1362–1540)
FEATURES             Location/Qualifiers
     source          1..6127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..17
                     /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             40..693
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     651..673
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             700..717
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             718..738
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1959..1981)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2126..2144)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2212..2316
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2317..2385,2386..3177)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2535..2554)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3348..3936
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3837..3856
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4090..4107
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4180..4257
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4258..5340
                     /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(4277..4296)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5389..5394,5395..5412,5413..5419)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5427..5443
                     /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     5451..5467
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5451..5467
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5463..5990
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5483..5500)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5483..5499)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5492..5514)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(6092..6097,6098..6113,6114..6120)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
ORIGIN
        1 ttgtgagcgg ataacaattt cacacaggaa acagtattca tgtcccctat actaggttat
       61 tggaaaatta agggccttgt gcaacccact cgacttcttt tggaatatct tgaagaaaaa
      121 tatgaagagc atttgtatga gcgcgatgaa ggtgataaat ggcgaaacaa aaagtttgaa
      181 ttgggtttgg agtttcccaa tcttccttat tatattgatg gtgatgttaa attaacacag
      241 tctatggcca tcatacgtta tatagctgac aagcacaaca tgttgggtgg ttgtccaaaa
      301 gagcgtgcag agatttcaat gcttgaagga gcggttttgg atattagata cggtgtttcg
      361 agaattgcat atagtaaaga ctttgaaact ctcaaagttg attttcttag caagctacct
      421 gaaatgctga aaatgttcga agatcgttta tgtcataaaa catatttaaa tggtgatcat
      481 gtaacccatc ctgacttcat gttgtatgac gctcttgatg ttgttttata catggaccca
      541 atgtgcctgg atgcgttccc aaaattagtt tgttttaaaa aacgtattga agctatccca
      601 caaattgata agtacttgaa atccagcaag tatatagcat ggcctttgca gggctggcaa
      661 gccacgtttg gtggtggcga ccatcctcca aaatcggatc tggttccgcg tggatccgag
      721 aacctgtatt tccaaggctc caaaagtggt gctcagacac ccaaaagtcc acctgaacac
      781 tatgttcagg agaccccact catgtttagc agatgtactt ctgtcagttc acttgatagt
      841 tttgagagtc gttcgattgc cagctccgtt cagagtgaac catgcagtgg aatggtaagt
      901 ggcattataa gccccagtga tcttccagat agccctggac aaaccatgcc accaagcaga
      961 agtaaaacac ctccaccacc tcctcaaaca gctcaaacca agcgagaagt acctaaaaat
     1021 aaagcaccta ctgctgaaaa gagagagagt ggacctaagc aagctgcagt aaatgctgca
     1081 gttcagaggg tccaggttct tccagatgct gatactttat tacattttgc cacggaaagt
     1141 actccagatg gattttcttg ttcatccagc ctgagtgctc tgagcctcga tgagccattt
     1201 atacagaaag atgtggaatt aagaataatg cctccagttc aggaaaatga caatgggaat
     1261 gaaacagaat cagagtagcc taaagaatca aatgaaaacc aagagaaaga ggcagaaaaa
     1321 actattgatt ctgaaaagga cctattagat gattcagatg atgatgatat tgaaatacta
     1381 gaagaatgta ttatttctgc catgccaaca aagtcatcac gtaaagcaaa aaagccagcc
     1441 cagactgctt caaaattacc tccacctgtg gcaaggaaac caagtcagct gcctgtgtac
     1501 aaacttctac catcacaaaa caggttgcaa ccccaaaagc atgttagttt tacaccgggg
     1561 gatgatatgc cacgggtgta ttgtgttgaa gggacaccta taaacttttc cacagctaca
     1621 tctctaagtg atctaacaat cgaatcccct ccaaatgagt tagctgctgg agaaggagtt
     1681 agaggagggg cacagtcagg tgaatttgaa aaacgagata ccattcctac agaaggcaga
     1741 agtacagatg aggctcaagg aggaaaaacc tcatctgtaa ccatacctga attggatgac
     1801 aataaagcag aggaaggtga tattcttgca gaatgcatta attctgctat gcccaaaggg
     1861 aaaagtcaca agcctttccg tgtgaaaaag tagctcgagc ggccgcatcg tgactgactg
     1921 acgatctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     1981 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     2041 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     2101 gtgtataatt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     2161 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     2221 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     2281 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     2341 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     2401 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     2461 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     2521 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     2581 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     2641 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2701 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2761 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2821 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     2881 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2941 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3001 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     3061 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3121 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3181 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3241 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3301 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3361 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3421 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3481 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3541 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3601 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3661 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3721 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3781 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3841 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3901 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3961 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4021 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4081 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     4141 cttacgcatc tgtgcggtat ttcacaccgc ataaattccg acaccatcga atggcgcaaa
     4201 acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg
     4261 aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc
     4321 cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg
     4381 atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg
     4441 ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg
     4501 gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga
     4561 agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg
     4621 ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact
     4681 aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc
     4741 tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa
     4801 atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg
     4861 cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt
     4921 gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg
     4981 atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg
     5041 ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt
     5101 tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg
     5161 gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc
     5221 tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg
     5281 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     5341 gcgcaacgca attaatgtaa gttagctcac tcattaggca ccccaggctt tacactttat
     5401 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     5461 ctatgaccat gattacggat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     5521 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     5581 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
     5641 gctttgcctg gtttccggca ccagaagcgg tgccggaaag ctggctggag tgcgatcttc
     5701 ctgaggccga tactgtcgtc gtcccctcaa actggcagat gcacggttac gatgcgccca
     5761 tctacaccaa cgtgacctat cccattacgg tcaatccgcc gtttgttccc acggagaatc
     5821 cgacgggttg ttactcgctc acatttaatg ttgatgaaag ctggctacag gaaggccaga
     5881 cgcgaattat ttttgatggc gttggaatta gcttatcgac tgcacggtgc accaatgctt
     5941 ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat
     6001 aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata
     6061 acggttctgg caaatattct gaaatgagct gttgacaatt aatcatcggc tcgtataatg
     6121 tgtggaa
//
