LOCUS       sequence-322378-        4973 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control for beta-catenin reporter. Contains mutated TCF/LEF binding 
            sites upstream of a luciferase reporter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4973)
  AUTHORS   Veeman MT, Slusarski DC, Kaykas A, Louie SH, Moon RT
  TITLE     Zebrafish prickle, a modulator of noncanonical Wnt/Fz signaling, 
            regulates gastrulation movements.
  JOURNAL   Curr Biol. 2003 Apr 15. 13(8):680-5.
  PUBMED    12699626
REFERENCE   2  (bases 1 to 4973)
  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: "Curr Biol. 
            2003 Apr 15. 13(8):680-5."
COMMENT     Sequence Label: M51 Super 8x FOPFlash (TOPFlash mutant)
FEATURES             Location/Qualifiers
     source          1..4973
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    7..55
                     /note="synthetic polyadenylation signal"
     misc_feature    69..160
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     109..128
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        317..348
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      404..413
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             410..2062
                     /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(476..494)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2103..2224
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2114..2133)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2168..2187
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2472..2489)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2643..3231)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2723..2742)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3402..4193,4194..4262))
                     /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     4025..4044
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4263..4367)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4394..4849
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4481..4500)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4691..4712
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg
       61 atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag
      121 gctgtcccca gtgcaagtgc aggtgccaga acatttctct atcgataggt accttacgcg
      181 aggccaaagg gggtaaggcc aaagggggta aggccaaagg gggtaaggcc aaaggcgcga
      241 ggccaaaggg ggtaaggcca aagggggtaa ggccaaaggg ggtaaggcca aaggcccggg
      301 ctcgagatct agactctaga gggtatataa tggaagctcg aattccagct tggcattccg
      361 gtactgttgg taaaaagctt ggcattccgg tactgttggt aaagccacca tggaagacgc
      421 caaaaacata aagaaaggcc cggcgccatt ctatccgctg gaagatggaa ccgctggaga
      481 gcaactgcat aaggctatga agagatacgc cctggttcct ggaacaattg cttttacaga
      541 tgcacatatc gaggtggaca tcacttacgc tgagtacttc gaaatgtccg ttcggttggc
      601 agaagctatg aaacgatatg ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa
      661 ctctcttcaa ttctttatgc cggtgttggg cgcgttattt atcggagttg cagttgcgcc
      721 cgcgaacgac atttataatg aacgtgaatt gctcaacagt atgggcattt cgcagcctac
      781 cgtggtgttc gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa aaaagctccc
      841 aatcatccaa aaaattatta tcatggattc taaaacggat taccagggat ttcagtcgat
      901 gtacacgttc gtcacatctc atctacctcc cggttttaat gaatacgatt ttgtgccaga
      961 gtccttcgat agggacaaga caattgcact gatcatgaac tcctctggat ctactggtct
     1021 gcctaaaggt gtcgctctgc ctcatagaac tgcctgcgtg agattctcgc atgccagaga
     1081 tcctattttt ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg ttccattcca
     1141 tcacggtttt ggaatgttta ctacactcgg atatttgata tgtggatttc gagtcgtctt
     1201 aatgtataga tttgaagaag agctgtttct gaggagcctt caggattaca agattcaaag
     1261 tgcgctgctg gtgccaaccc tattctcctt cttcgccaaa agcactctga ttgacaaata
     1321 cgatttatct aatttacacg aaattgcttc tggtggcgct cccctctcta aggaagtcgg
     1381 ggaagcggtt gccaagaggt tccatctgcc aggtatcagg caaggatatg ggctcactga
     1441 gactacatca gctattctga ttacacccga gggggatgat aaaccgggcg cggtcggtaa
     1501 agttgttcca ttttttgaag cgaaggttgt ggatctggat accgggaaaa cgctgggcgt
     1561 taatcaaaga ggcgaactgt gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa
     1621 tccggaagcg accaacgcct tgattgacaa ggatggatgg ctacattctg gagacatagc
     1681 ttactgggac gaagacgaac acttcttcat cgttgaccgc ctgaagtctc tgattaagta
     1741 caaaggctat caggtggctc ccgctgaatt ggaatccatc ttgctccaac accccaacat
     1801 cttcgacgca ggtgtcgcag gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt
     1861 tgttgttttg gagcacggaa agacgatgac ggaaaaagag atcgtggatt acgtcgccag
     1921 tcaagtaaca accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa
     1981 aggtcttacc ggaaaactcg acgcaagaaa aatcagagag atcctcataa aggccaagaa
     2041 gggcggaaag atcgccgtgt aattctagag tcggggcggc cggccgcttc gagcagacat
     2101 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2161 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2221 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     2281 tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtcgaccga
     2341 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg
     2401 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc
     2461 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     2521 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2581 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2641 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2701 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2761 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2821 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2881 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2941 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3001 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3061 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3121 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3181 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3241 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3301 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3361 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     3421 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     3481 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3541 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3601 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3661 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3721 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3781 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3841 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3901 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     3961 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     4021 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4081 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4141 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     4201 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     4261 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     4321 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     4381 aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4441 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4501 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4561 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4621 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4681 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4741 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4801 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttg ccattcgcca
     4861 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     4921 cccaagctac catgataagt aagtaatatt aaggtacggg aggtacttgg agc
//
