LOCUS       sequence-378722-        8613 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to test the Zebrafish Wnt3 promoter activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8613)
  AUTHORS   Vogg MC, Beccari L, Iglesias Olle L, Rampon C, Vriz S, Perruchoud C,
            Wenger Y, Galliot B
  TITLE     An evolutionarily-conserved Wnt3/beta-catenin/Sp5 feedback loop 
            restricts head organizer activity in Hydra.
  JOURNAL   Nat Commun. 2019 Jan 18;10(1):312. doi: 10.1038/s41467-018-08242-2.
  PUBMED    30659200
REFERENCE   2  (bases 1 to 8613)
  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: "Nat 
            Commun."; date: "2019-01-18"; volume: "10(1):312. doi"; pages: " 
            10.1038/s41467-018-08242-2"
COMMENT     Sequence Label: ZfWnt3:Luc
FEATURES             Location/Qualifiers
     source          1..8613
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      30..39
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             36..1688
                     /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(102..120)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1729..1850
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1740..1759)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1794..1813
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2098..2115)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2269..2857)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2349..2368)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3028..3819,3820..3888))
                     /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     3651..3670
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3889..3993)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4020..4475
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4107..4126)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4317..4338
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aagcttggca ttccggtact gttggtaaag ccaccatgga agacgccaaa aacataaaga
       61 aaggcccggc gccattctat ccgctggaag atggaaccgc tggagagcaa ctgcataagg
      121 ctatgaagag atacgccctg gttcctggaa caattgcttt tacagatgca catatcgagg
      181 tggacatcac ttacgctgag tacttcgaaa tgtccgttcg gttggcagaa gctatgaaac
      241 gatatgggct gaatacaaat cacagaatcg tcgtatgcag tgaaaactct cttcaattct
      301 ttatgccggt gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg aacgacattt
      361 ataatgaacg tgaattgctc aacagtatgg gcatttcgca gcctaccgtg gtgttcgttt
      421 ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa gctcccaatc atccaaaaaa
      481 ttattatcat ggattctaaa acggattacc agggatttca gtcgatgtac acgttcgtca
      541 catctcatct acctcccggt tttaatgaat acgattttgt gccagagtcc ttcgataggg
      601 acaagacaat tgcactgatc atgaactcct ctggatctac tggtctgcct aaaggtgtcg
      661 ctctgcctca tagaactgcc tgcgtgagat tctcgcatgc cagagatcct atttttggca
      721 atcaaatcat tccggatact gcgattttaa gtgttgttcc attccatcac ggttttggaa
      781 tgtttactac actcggatat ttgatatgtg gatttcgagt cgtcttaatg tatagatttg
      841 aagaagagct gtttctgagg agccttcagg attacaagat tcaaagtgcg ctgctggtgc
      901 caaccctatt ctccttcttc gccaaaagca ctctgattga caaatacgat ttatctaatt
      961 tacacgaaat tgcttctggt ggcgctcccc tctctaagga agtcggggaa gcggttgcca
     1021 agaggttcca tctgccaggt atcaggcaag gatatgggct cactgagact acatcagcta
     1081 ttctgattac acccgagggg gatgataaac cgggcgcggt cggtaaagtt gttccatttt
     1141 ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct gggcgttaat caaagaggcg
     1201 aactgtgtgt gagaggtcct atgattatgt ccggttatgt aaacaatccg gaagcgacca
     1261 acgccttgat tgacaaggat ggatggctac attctggaga catagcttac tgggacgaag
     1321 acgaacactt cttcatcgtt gaccgcctga agtctctgat taagtacaaa ggctatcagg
     1381 tggctcccgc tgaattggaa tccatcttgc tccaacaccc caacatcttc gacgcaggtg
     1441 tcgcaggtct tcccgacgat gacgccggtg aacttcccgc cgccgttgtt gttttggagc
     1501 acggaaagac gatgacggaa aaagagatcg tggattacgt cgccagtcaa gtaacaaccg
     1561 cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt accgaaaggt cttaccggaa
     1621 aactcgacgc aagaaaaatc agagagatcc tcataaaggc caagaagggc ggaaagatcg
     1681 ccgtgtaatt ctagagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg
     1741 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     1801 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1861 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     1921 aaaacctcta caaatgtggt aaaatcgata aggatccgtc gaccgatgcc cttgagagcc
     1981 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg
     2041 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc
     2101 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     2161 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2221 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2281 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2341 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2401 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2461 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2521 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2581 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2641 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2701 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2761 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2821 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2881 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2941 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3001 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3061 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3121 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     3181 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3241 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3301 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3361 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3421 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3481 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3541 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     3601 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     3661 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     3721 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     3781 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     3841 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     3901 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     3961 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     4021 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4081 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4141 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4201 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4261 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4321 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4381 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4441 cgcgaatttt aacaaaatat taacgcttac aatttgccat tcgccattca ggctgcgcaa
     4501 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagccca agctaccatg
     4561 ataagtaagt aatattaagg tacgggaggt acttggagcg gccgctaaac tatcaataaa
     4621 agaagtgtct gttgtggggc cgagggtcct ggggggctgc ggagctttag gcattgtcct
     4681 ggagggtgat tgagagcgag aggaggggag ccgagaagca ttgtgcgagg acggagcctt
     4741 tctaaaggtg gttattagga gagaaatggt gtgggggtgg ctgttgactc ttcatcttgt
     4801 gtgaacaaag tatctggcgt atagctttct ctctctctct ctcgctctct tgcggctttc
     4861 gctttacgac ggccatgaat actctcggcg ggctcgccca gacctgcgcg aaggcttatt
     4921 gtccggctta agtgcggttt aattaagcga gctctgtgtc ctgagagaag agaggagctt
     4981 tagagcgctc gggtcctgcg gagatgcaga ggatgatgcg taatatcagg gctattgaag
     5041 gggaaagtgt ttgcggctct aatgaaaaag gtttgtttgg gggggatgag tggggagggg
     5101 attgaggagt gtctggcctt tgtctccagc ttacaaagta gggacaaaag cagtgatatt
     5161 atacaaatgc acccatttcc aggacatgat gcgaggacgg gggcctggag atgggtcgga
     5221 ctgctccatc tcagccccgt tggcacctta caaaatgatt ttttcccccc accacaaaga
     5281 taggcattgg ggataaaccc cgagagctgg ggcttctggg taccgcaaat ttgggggttg
     5341 tggacttaag cggattttct ctgcaactgt aagaacaagc taattcctca gacctccttt
     5401 atctctaaca tggtaataac aaacaatgaa attctgaaga gaaaggggca aaggaagagg
     5461 agggaggaga gacgcagaca caaaagaagg agagatggag ggaatgaaag agaggagaga
     5521 cgtgtgtgag gagagggtgg gcttgggcgt gttaagaaac tttctcgctc tagtgttatg
     5581 tgtgaacact ctgctaatcc aggttttatt gcttgcctgt gtccggccac actgtccgga
     5641 ttgagtgtgt gtcagggata gtgggggtct acacatgtcc ctctttgagg aaagcctgag
     5701 aaagagccgg acctggttac ggccgttttg gggtcgccta agaaacctgc accacagggg
     5761 tcctctgtgt atgtctctga attcaacgtg catgttgcta gcaaacacat acggattaat
     5821 gctaatgatg aaggtgtggc tatattaggt acaaaaaaaa actttgacaa tagttgtata
     5881 aactcctgtc aacagtccat caatgtatga agcagaactc gctttgaaac caaacattgt
     5941 tgtgtgggtc catttggacc agtttaagga aatcttttgc cctataaatt ccagcctgca
     6001 tgaattttta tgattattgc tcataaacat acttttaaaa atgtgcattt taactaacta
     6061 tcaacttgtg cccgtccact caaaggaagc caaaagggtg tctctcccag atcacaggga
     6121 ctaccttgtg ggtgtgtagg gtgatgtatt agctaggcgc taataaaatg caggagtgtc
     6181 cacaaacatg acttaatgag ggaggatgag agactcctga cccagaggtc aaagctccgg
     6241 gcgactctct ttattcccat tgatctgctg gtgcacttca tttccatgct ctagggccca
     6301 cttcagtaaa ttcattaagt gcgattgata cgagaatcaa gacttcctcc aaaacacaat
     6361 ggctatgcta ggagggaatg gtcttgtttg tgctgcagct atgtcaagca tctctggctt
     6421 ttgtcctggt ttcgcaccat tgcggtgcat gctggggttt gagtcctcga gattctaatc
     6481 gtaagctcat ttccagctgg ttcgttgtgt tgcgccccta tggccttgac ctgggaaacg
     6541 tggtttcagt gttgttgaga tgccgaccgc agaccgttcc ttgcctccac tcgtaaatta
     6601 cgctccgatc aggttttccc agccaatcct ggcgagcaga tctgcatctg tcactctgca
     6661 agaatgctga atcaaaccgc tgatctaggc acaatacagc tgatggaaag cagatcttta
     6721 ttccaacatc aggaacgtag cacgagccga gtgcttctgg aagctgcaat ttaaccaaca
     6781 ggttcatatc tgatgagatc aaggatgcat catcctggaa gaaacactgt gatgaacatt
     6841 taagatgtca gaatgacccg tcatgcctct gagctgagtc cccgcacagc gcttattctt
     6901 ttgaagaggg agaataaaag caaaaaaaca gaggcgatgg aaatatgaca tccagaccgc
     6961 ggtcagaact gtgagtgtct ctcctgggct cctatggttg tggtgcacag tccatgtccc
     7021 tgctatgagt tagatagaca gatgcactgg cagatagaca gacagacagg ctttactact
     7081 tggtcgctgg attgcaggaa agagctttat ccaaccagag tcaagcaagc aacatgggac
     7141 aataacataa ttaaatattc tggggaaaag tggcacttat gcaaatgttt tttgtaatgg
     7201 gttggagaaa attggccgga acctacataa tttttaatga caggatgatt catttgtgaa
     7261 attgtaattc aaagaaattc agtaaactaa ctaacaatct aaaaaattct ttgatttgca
     7321 gtttaattct acttcctaac attgatttgt ttcaacttta ttcaaatggc aaaactaaat
     7381 tagtactcaa agctccaaat gatatgattc ctagactgtt taaagtgcta aaagtgccaa
     7441 aacttacagc tacccacatg aacttatatt tcattctgcc tccatttgca attcctccag
     7501 tgtttgtacc tgaaactgtt gtcagttctc acctctgtcc tgccgttcat tctgacacag
     7561 agactctata tttgaatgta ttgtcaactt ctaatgccgt gtctactact ctgtgctgtc
     7621 tgctctcatc ccatccgaat gacaatagtg tttatgtaaa atgtgctgcc tttgggccgc
     7681 tggggcacag aacactgcat gcctggccct gccacaccac cttttacggc tttttattgt
     7741 gttgcctttg tgtcacaagg ggaggacgcg gccatcaaaa ttctccaaga caaaattctt
     7801 gaatgtatgt gcacgcacac gcatccggaa caattagttc aggagcacgg gtgcttaaaa
     7861 aacggagacg acacttctcg ccaccacctc ctcgccgagc agcaggcccg agtctggcac
     7921 agctgggttt ccttgatgct aacgcagaca atatagatct taataagaca aatctacctc
     7981 tccgaaactc agtcctgaca ccctttcatt cttattccct cccagcctcc tctctctctc
     8041 tctctctctt tctttctttc tttgctacat taccaggggg tggtgcatga ttgacagatg
     8101 ggcaggaata ataatttttt tttttttttt gcgtaaccta aggccatgct ctgtgattgg
     8161 cttgccagct gacatctcca aacctctctt gtcgtttttt tcccctctct ctctctcgct
     8221 ttctctctct cactctctct ctctaatatt ggccctgatg atcctaaggg gaaaaaagtg
     8281 gggagagaaa aagatatgta ccttagggaa gaacctcagt aagagggagg gaatcagagg
     8341 acgagtagga gcgtgtgtgc gagtgtgtga aagagacact tactctccta actactactg
     8401 gaagacacac acttttctca cccaactcaa agcagcagca aaccttggag aattgtttta
     8461 tttttgtttt ttcgcgaacc cccctaaaaa aattcctaat tattctctca agcatccatc
     8521 tttgccttgc cgggatttaa ttttcacccc ctctctagga atcctatgta ctgatcaata
     8581 gaccctttcc atccacacaa gacatacctc ccc
//
