LOCUS       sequence-152179-        4914 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4914)
  AUTHORS   Shih IM, Yu J, He TC, Vogelstein B, Kinzler KW
  TITLE     The beta-catenin binding domain of adenomatous polyposis coli is 
            sufficient for tumor suppression.
  JOURNAL   Cancer Res. 2000 Mar 15. 60(6):1671-6.
  PUBMED    10749138
REFERENCE   2  (bases 1 to 4914)
  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: "Cancer Res.
            2000 Mar 15. 60(6):1671-6."
COMMENT     Sequence Label: pGL3-OF (mutant TCF-4 sites)
FEATURES             Location/Qualifiers
     source          1..4914
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    98..146
                     /note="synthetic polyadenylation signal"
     misc_feature    160..251
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     200..219
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      436..445
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             442..2094
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(508..526)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2135..2256
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2146..2165)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2200..2219
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2504..2521)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2675..3263)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2755..2774)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3434..4225,4226..4294))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4057..4076
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4295..4399)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4426..4881
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4513..4532)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4723..4744
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aagggcgatc ggtgcgggcc tcttcgctat tacgccagcc caagctacca tgataagtaa
       61 gtaatattaa ggtacgggag gtacttggag cggccgcaat aaaatatctt tattttcatt
      121 acatctgtgt gttggttttt tgtgtgaatc gatagtacta acatacgctc tccatcaaaa
      181 caaaacgaaa caaaacaaac tagcaaaata ggctgtcccc agtgcaagtg caggtgccag
      241 aacatttctc tatcgatagg taccgagctc ttacgcgtgc tagcccgggc tcgagcagct
      301 gaagcttgca tgcctgcagg gccaaagggg gtaaggccaa agggggtaag gccaaaggtc
      361 tagagggtat ataatggatc cggtactcga gatctgcgat ctaagtaagc ttggcattcc
      421 ggtactgttg gtaaagccac catggaagac gccaaaaaca taaagaaagg cccggcgcca
      481 ttctatccgc tggaagatgg aaccgctgga gagcaactgc ataaggctat gaagagatac
      541 gccctggttc ctggaacaat tgcttttaca gatgcacata tcgaggtgga catcacttac
      601 gctgagtact tcgaaatgtc cgttcggttg gcagaagcta tgaaacgata tgggctgaat
      661 acaaatcaca gaatcgtcgt atgcagtgaa aactctcttc aattctttat gccggtgttg
      721 ggcgcgttat ttatcggagt tgcagttgcg cccgcgaacg acatttataa tgaacgtgaa
      781 ttgctcaaca gtatgggcat ttcgcagcct accgtggtgt tcgtttccaa aaaggggttg
      841 caaaaaattt tgaacgtgca aaaaaagctc ccaatcatcc aaaaaattat tatcatggat
      901 tctaaaacgg attaccaggg atttcagtcg atgtacacgt tcgtcacatc tcatctacct
      961 cccggtttta atgaatacga ttttgtgcca gagtccttcg atagggacaa gacaattgca
     1021 ctgatcatga actcctctgg atctactggt ctgcctaaag gtgtcgctct gcctcataga
     1081 actgcctgcg tgagattctc gcatgccaga gatcctattt ttggcaatca aatcattccg
     1141 gatactgcga ttttaagtgt tgttccattc catcacggtt ttggaatgtt tactacactc
     1201 ggatatttga tatgtggatt tcgagtcgtc ttaatgtata gatttgaaga agagctgttt
     1261 ctgaggagcc ttcaggatta caagattcaa agtgcgctgc tggtgccaac cctattctcc
     1321 ttcttcgcca aaagcactct gattgacaaa tacgatttat ctaatttaca cgaaattgct
     1381 tctggtggcg ctcccctctc taaggaagtc ggggaagcgg ttgccaagag gttccatctg
     1441 ccaggtatca ggcaaggata tgggctcact gagactacat cagctattct gattacaccc
     1501 gagggggatg ataaaccggg cgcggtcggt aaagttgttc cattttttga agcgaaggtt
     1561 gtggatctgg ataccgggaa aacgctgggc gttaatcaaa gaggcgaact gtgtgtgaga
     1621 ggtcctatga ttatgtccgg ttatgtaaac aatccggaag cgaccaacgc cttgattgac
     1681 aaggatggat ggctacattc tggagacata gcttactggg acgaagacga acacttcttc
     1741 atcgttgacc gcctgaagtc tctgattaag tacaaaggct atcaggtggc tcccgctgaa
     1801 ttggaatcca tcttgctcca acaccccaac atcttcgacg caggtgtcgc aggtcttccc
     1861 gacgatgacg ccggtgaact tcccgccgcc gttgttgttt tggagcacgg aaagacgatg
     1921 acggaaaaag agatcgtgga ttacgtcgcc agtcaagtaa caaccgcgaa aaagttgcgc
     1981 ggaggagttg tgtttgtgga cgaagtaccg aaaggtctta ccggaaaact cgacgcaaga
     2041 aaaatcagag agatcctcat aaaggccaag aagggcggaa agatcgccgt gtaattctag
     2101 agtcggggcg gccggccgct tcgagcagac atgataagat acattgatga gtttggacaa
     2161 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     2221 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     2281 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     2341 tgtggtaaaa tcgataagga tccgtcgacc gatgcccttg agagccttca acccagtcag
     2401 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat
     2461 catgcaactc gtaggacagg tgccggcagc gctcttccgc ttcctcgctc actgactcgc
     2521 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2581 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2641 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2701 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2761 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2821 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2881 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2941 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3001 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3061 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3121 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3181 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3241 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3301 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3361 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3421 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3481 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3541 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3601 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3661 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3721 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3781 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3841 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3901 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3961 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4021 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4081 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4141 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4201 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4261 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4321 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4381 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg
     4441 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4501 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4561 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4621 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     4681 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4741 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     4801 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     4861 aaatattaac gcttacaatt tgccattcgc cattcaggct gcgcaactgt tggg
//
