LOCUS       sequence-259650-        5452 bp    DNA     circular SYN 15-APR-2025
DEFINITION  expresses human APC-B,C (1133–1189) in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5452)
  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 5452)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-B,C (1133–1189)
FEATURES             Location/Qualifiers
     source          1..5452
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(600..605,606..623,624..630)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    638..654
                     /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     662..678
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     662..678
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             674..1201
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(694..711)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(694..710)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(703..725)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1303..1308,1309..1324,1325..1331)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1339..1355
                     /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             1378..2031
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1989..2011
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2038..2055
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2056..2076
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2622..2644)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2789..2807)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2875..2979
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2980..3048,3049..3840)
                     /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(3198..3217)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4011..4599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4500..4519
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4753..4770
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4843..4920
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(4921..5452,1..551)
                     /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(4940..4959)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc
       61 tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag
      121 gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg
      181 ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta
      241 ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag
      301 acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc
      361 aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc
      421 tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct
      481 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
      541 gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc accccaggct
      601 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
      661 acaggaaaca gctatgacca tgattacgga ttcactggcc gtcgttttac aacgtcgtga
      721 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
      781 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
      841 tggcgaatgg cgctttgcct ggtttccggc accagaagcg gtgccggaaa gctggctgga
      901 gtgcgatctt cctgaggccg atactgtcgt cgtcccctca aactggcaga tgcacggtta
      961 cgatgcgccc atctacacca acgtgaccta tcccattacg gtcaatccgc cgtttgttcc
     1021 cacggagaat ccgacgggtt gttactcgct cacatttaat gttgatgaaa gctggctaca
     1081 ggaaggccag acgcgaatta tttttgatgg cgttggaatt agcttatcga ctgcacggtg
     1141 caccaatgct tctggcgtca ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta
     1201 aatcactgca taattcgtgt cgctcaaggc gcactcccgt tctggataat gttttttgcg
     1261 ccgacatcat aacggttctg gcaaatattc tgaaatgagc tgttgacaat taatcatcgg
     1321 ctcgtataat gtgtggaatt gtgagcggat aacaatttca cacaggaaac agtattcatg
     1381 tcccctatac taggttattg gaaaattaag ggccttgtgc aacccactcg acttcttttg
     1441 gaatatcttg aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg tgataaatgg
     1501 cgaaacaaaa agtttgaatt gggtttggag tttcccaatc ttccttatta tattgatggt
     1561 gatgttaaat taacacagtc tatggccatc atacgttata tagctgacaa gcacaacatg
     1621 ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc ggttttggat
     1681 attagatacg gtgtttcgag aattgcatat agtaaagact ttgaaactct caaagttgat
     1741 tttcttagca agctacctga aatgctgaaa atgttcgaag atcgtttatg tcataaaaca
     1801 tatttaaatg gtgatcatgt aacccatcct gacttcatgt tgtatgacgc tcttgatgtt
     1861 gttttataca tggacccaat gtgcctggat gcgttcccaa aattagtttg ttttaaaaaa
     1921 cgtattgaag ctatcccaca aattgataag tacttgaaat ccagcaagta tatagcatgg
     1981 cctttgcagg gctggcaagc cacgtttggt ggtggcgacc atcctccaaa atcggatctg
     2041 gttccgcgtg gatccgagaa cctgtatttc caaggcgatg actatgaaga tgataagcct
     2101 accaattata gtgaacgtta ctctgaagaa gaacagcatg aagaagaaga gagaccaaca
     2161 aattatagca taaaatataa tgaagagaaa cgtcatgtgg atcagcctat tgattatagt
     2221 ttaaaatatg ccacagatat tccttcatga cagaaacagt cattttcatt ctcaaagagt
     2281 tcatctggac aaagcagtaa aaccgaacat atgtcttcaa gcagtgagaa tacgtccaca
     2341 ccttcatcta atgccaagag gcagaatcag ctccatccaa gttctgcaca gagtagaagt
     2401 ggtcagcctc aaaaggctgc cacttgcaaa gtttcttcta ttaaccaaga aacaatacag
     2461 acttattgtg tagaagatac tccaatatgt ttttcaagat gtagttcatt atcatctttg
     2521 tcatcagctg aagatgaaat aggatgtaat cagtagctcg agcggccgca tcgtgactga
     2581 ctgacgatct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     2641 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     2701 gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc
     2761 ggagtgtata attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     2821 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     2881 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2941 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     3001 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     3061 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     3121 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     3181 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     3241 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     3301 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     3361 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     3421 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     3481 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
     3541 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3601 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3661 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3721 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3781 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3841 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     3901 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3961 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4021 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4081 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4141 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     4201 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4261 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4321 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4381 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4441 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4501 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4561 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4621 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     4681 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     4741 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     4801 ctccttacgc atctgtgcgg tatttcacac cgcataaatt ccgacaccat cgaatggcgc
     4861 aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat
     4921 gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt
     4981 tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg
     5041 gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag
     5101 tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc
     5161 gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa
     5221 cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt
     5281 gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc
     5341 actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt
     5401 ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt gg
//
