LOCUS       sequence-378726-        6555 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to test the Hydra Wnt3 promoter activity (full deletion of 
            the repressor element).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6555)
  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 6555)
  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: HyWnt3-Delta-Rep:Luc
FEATURES             Location/Qualifiers
     source          1..6555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1819..1828
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1825..3477
                     /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(1891..1909)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3518..3639
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3529..3548)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3583..3602
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3887..3904)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4058..4646)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4138..4157)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4817..5608,5609..5677))
                     /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     5440..5459
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5678..5782)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5809..6264
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5896..5915)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6106..6127
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6395..6443
                     /note="synthetic polyadenylation signal"
     misc_feature    6457..6548
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6497..6516
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagt tgaagagcaa gaaaacgaat gattgtaaat tgtacatgtc gattgtgaaa
       61 attgactata aataattatt aaaaatgtag attttatgtg cgaggaaaac atgaagacca
      121 tagaaaactc caaagcactg atgttcactg aaaaaactag ataaataaaa cgattttaac
      181 taagaggaac agtaaaaaga tgcaacttag ctaaatgatg agtaacgcga aatagagttt
      241 tggttgatgg aaaaaatatc gatcgctaac ttgaacagcg aacatgtagt ttttgttgct
      301 actggatacc aaaccatggt aataatgcta tccttttttt taattttaat tttgactatt
      361 tttttaaatc ttaagcccgc gctctccatc ggtaaaaaag aagacaatgg caaccctaaa
      421 ttaatgtatg tatgtaaaag gtcatcatat attaacatat tataataaaa tattatttat
      481 gttttaatac tatttcttat attttttgat tttcataaaa aatgatttgt tttttataga
      541 ctactattat tatcatatta tttttatatt tcatttttat gtttttatta ctaatataac
      601 tattattaat ctaaaacagt ggaataaaca tttagaatat tgttgacttc atattcgcct
      661 aaaaagcata tgtttacatt ttttattaag taattatctt tgtttggcat ttaataatat
      721 taaaaagtat tcgtcaccaa aactctgctt tttcaaattt ataaatttat gttaacgtta
      781 aaacaatgca tcaatgcatt gaacgttttt ttaatggaaa aattaacaat tttactatct
      841 ttataagttt gtttagtttc aagttttata taatgtagct tgagtgtaaa aatattattt
      901 gagtctaaca ataaatctaa aataactttt tgttgatgct agattttata catctttaac
      961 agctatggcg ccaaaagaga cagcgggtgt tttattgagc tacgcagtta ttgttttttt
     1021 aaattcagtc tttatcaaat gttaacaaat aaaaatcctg attgcatccc aatcagctca
     1081 tttggaatat gttatgacca accaaattgt tggcaaagta gttgtaaatt atgatgtcat
     1141 cttcttatag gaaaatgaca tcagtaaata atcgtcagaa aaaaacgtca gaaaacaagt
     1201 gacgtattat aattttgttt ataatataca tatatatata taaaactcat cgatgacaaa
     1261 acaatgacca ataaaatatt cctttgatag agtctcctaa tatgttatct tggtaatttg
     1321 tacaattatt ttattaaatg tttatcaatt aatgataaat ttaaacttat agagatattt
     1381 gcaatgttga taaaaaagat gtcgataaca tattaacgaa atagtatatg aatataaaca
     1441 tataatatta tcatatatat tatgcacatc aagttttatg tttactgacg tttccctgcc
     1501 ttctaactcc tacctgcaca aaaagcttga acttttatat tttttaatta gtataagcaa
     1561 atagacgtct taaagttaaa tcgatacttc tttaaaatct ttacaaacat taccatgtga
     1621 aatatttggc ttaatatatc agaacagcta tatcaatttc actgtaattt tatcgcactg
     1681 tgcttattat tagatattca ataacccatt gtgaaaaggc ttaaacccgt ttttactttg
     1741 ttgctttaaa ggtttgattt aactaaccgc cgagatctgc gatctaagta agcttggcat
     1801 tccggtactg ttggtaaagc caccatggaa gacgccaaaa acataaagaa aggcccggcg
     1861 ccattctatc cgctggaaga tggaaccgct ggagagcaac tgcataaggc tatgaagaga
     1921 tacgccctgg ttcctggaac aattgctttt acagatgcac atatcgaggt ggacatcact
     1981 tacgctgagt acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg atatgggctg
     2041 aatacaaatc acagaatcgt cgtatgcagt gaaaactctc ttcaattctt tatgccggtg
     2101 ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga acgacattta taatgaacgt
     2161 gaattgctca acagtatggg catttcgcag cctaccgtgg tgttcgtttc caaaaagggg
     2221 ttgcaaaaaa ttttgaacgt gcaaaaaaag ctcccaatca tccaaaaaat tattatcatg
     2281 gattctaaaa cggattacca gggatttcag tcgatgtaca cgttcgtcac atctcatcta
     2341 cctcccggtt ttaatgaata cgattttgtg ccagagtcct tcgataggga caagacaatt
     2401 gcactgatca tgaactcctc tggatctact ggtctgccta aaggtgtcgc tctgcctcat
     2461 agaactgcct gcgtgagatt ctcgcatgcc agagatccta tttttggcaa tcaaatcatt
     2521 ccggatactg cgattttaag tgttgttcca ttccatcacg gttttggaat gtttactaca
     2581 ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt atagatttga agaagagctg
     2641 tttctgagga gccttcagga ttacaagatt caaagtgcgc tgctggtgcc aaccctattc
     2701 tccttcttcg ccaaaagcac tctgattgac aaatacgatt tatctaattt acacgaaatt
     2761 gcttctggtg gcgctcccct ctctaaggaa gtcggggaag cggttgccaa gaggttccat
     2821 ctgccaggta tcaggcaagg atatgggctc actgagacta catcagctat tctgattaca
     2881 cccgaggggg atgataaacc gggcgcggtc ggtaaagttg ttccattttt tgaagcgaag
     2941 gttgtggatc tggataccgg gaaaacgctg ggcgttaatc aaagaggcga actgtgtgtg
     3001 agaggtccta tgattatgtc cggttatgta aacaatccgg aagcgaccaa cgccttgatt
     3061 gacaaggatg gatggctaca ttctggagac atagcttact gggacgaaga cgaacacttc
     3121 ttcatcgttg accgcctgaa gtctctgatt aagtacaaag gctatcaggt ggctcccgct
     3181 gaattggaat ccatcttgct ccaacacccc aacatcttcg acgcaggtgt cgcaggtctt
     3241 cccgacgatg acgccggtga acttcccgcc gccgttgttg ttttggagca cggaaagacg
     3301 atgacggaaa aagagatcgt ggattacgtc gccagtcaag taacaaccgc gaaaaagttg
     3361 cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa actcgacgca
     3421 agaaaaatca gagagatcct cataaaggcc aagaagggcg gaaagatcgc cgtgtaattc
     3481 tagagtcggg gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga
     3541 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     3601 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     3661 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     3721 aaatgtggta aaatcgataa ggatccgtcg accgatgccc ttgagagcct tcaacccagt
     3781 cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt
     3841 tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc cgcttcctcg ctcactgact
     3901 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3961 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4021 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4081 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4141 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4201 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4261 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4321 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4381 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4441 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4501 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4561 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4621 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4681 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4741 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4801 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4861 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4921 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4981 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5041 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5101 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5161 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5221 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5281 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5341 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5401 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5461 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5521 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5581 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5641 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5701 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5761 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta
     5821 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5881 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     5941 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     6001 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     6061 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     6121 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     6181 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     6241 acaaaatatt aacgcttaca atttgccatt cgccattcag gctgcgcaac tgttgggaag
     6301 ggcgatcggt gcgggcctct tcgctattac gccagcccaa gctaccatga taagtaagta
     6361 atattaaggt acgggaggta cttggagcgg ccgcaataaa atatctttat tttcattaca
     6421 tctgtgtgtt ggttttttgt gtgaatcgat agtactaaca tacgctctcc atcaaaacaa
     6481 aacgaaacaa aacaaactag caaaataggc tgtccccagt gcaagtgcag gtgccagaac
     6541 atttctctat cgata
//
