LOCUS       sequence-385730-        6657 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to test the Hydra Wnt3 promoter activity (partial deletion 
            of the repressor element-D2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6657)
  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 6657)
  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-D2:Luc
FEATURES             Location/Qualifiers
     source          1..6657
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1922..1931
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1928..3580
                     /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(1994..2012)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3621..3742
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3632..3651)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3686..3705
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3990..4007)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4161..4749)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4241..4260)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4920..5711,5712..5780))
                     /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     5543..5562
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5781..5885)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5912..6367
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5999..6018)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6209..6230
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6498..6546
                     /note="synthetic polyadenylation signal"
     misc_feature    6560..6651
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6600..6619
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 aggtaccgag ttgaagagca agaaaacgaa tgattgtaaa ttgtacatgt cgattgtgaa
       61 aattgactat aaataattat taaaaatgta gattttatgt gcgaggaaaa catgaagacc
      121 atagaaaact ccaaagcact gatgttcact gaaaaaacta gataaataaa acgattttaa
      181 ctaagaggaa cagtaaaaag atgcaactta gctaaatgat gagtaacgcg aaatagagtt
      241 ttggttgatg gaaaaaatat cgatcgctaa cttgaacagc gaacatgtag tttttgttgc
      301 tactggatac caaaccatgg taataatgct atcctttttt ttaattttaa ttttgactat
      361 ttttttaaat cttaagcccg cgctctccat cggtaaaaaa gaagacaatg gcaaccctaa
      421 attaatgtat gtatgtaaaa ggtcatcata tattaacata ttataataaa atattattta
      481 tgttttaata ctatttctta tattttttga ttttcataaa aaatgatttg ttttttatag
      541 actactatta ttatcatatt atttttatat ttcattttta tgtttttatt actaatataa
      601 ctattattaa tctaaaacag tggaataaac atttagaata ttgttgactt catattcgcc
      661 taaaaagcat atgtttacat tttttattaa gtaattatct ttgtttggca tttaataata
      721 ttaaaaagta ttcgtcacca aaactctgct ttttcaaatt tataaattta tgttaacgtt
      781 aaaacaatgc atcaatgcat tgaacgtttt tttaatggaa aaattaacaa ttttactatc
      841 tttataagtt tgtttagttt caagttttat ataatgtagc ttgagtgtaa aaatattatt
      901 tgagtctaac aataaatcta aaataacttt ttgttgatgc tagattttat acatctttaa
      961 cagctatggc gccaaaagag acagcgggtg ttttattgag ctacgcagtt attgtttttt
     1021 taaattcagt ctttatcaaa tgttaacaaa taaaaatcct gattgcatcc caatcagctc
     1081 atttggaata tgttatgacc aaccaaattg ttggcaaagt agttgtaaat tatgatgtca
     1141 tcttcttata ggaaaatgac atcagtaaat aatcgtcaga aaaaaacgtc agaaaacaag
     1201 tgacgtatta taattttgtt tataatatac atatatatat ataaaactca tcgatgacaa
     1261 aacaatgacc aataaaatat tcctttgata gagtctccta atatgttatc ttggtaattt
     1321 gtacaattat tttattaaat gtttatcaat taatgataaa tttaaactta tagagatatt
     1381 tgcaatgttg ataaaaaaga tgtcgataac atattaacga aatagtatat gaatataaac
     1441 atataatatt atcatatata ttatgcacat caagttttat gtttactgac gtttccctgc
     1501 cttctaactc ctacctgcac aaaaagcttg aacttttata ttttttaatt agtataagca
     1561 aatagacgtc ttaaagttaa atcgatactt ctttaaaatc tttacaaaca ttaccatgtg
     1621 aaatatttgg cttaatatat cagaacagct atatcaattt cactgtaatt ttatcgcact
     1681 gtgcttatta ttagatattc aataacccat tgtgaaaagg cttaaacccg tttttacttt
     1741 gttgctttaa aggtttgatt taactaaccg ctaatcatta taaaaagtta taaatttata
     1801 aaaatttatc tctgcaaaaa aagatttaac attaacacaa ctactacaaa aatctatctc
     1861 aggatactgt cctcgagatc tgcgatctaa gtaagcttgg cattccggta ctgttggtaa
     1921 agccaccatg gaagacgcca aaaacataaa gaaaggcccg gcgccattct atccgctgga
     1981 agatggaacc gctggagagc aactgcataa ggctatgaag agatacgccc tggttcctgg
     2041 aacaattgct tttacagatg cacatatcga ggtggacatc acttacgctg agtacttcga
     2101 aatgtccgtt cggttggcag aagctatgaa acgatatggg ctgaatacaa atcacagaat
     2161 cgtcgtatgc agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg cgttatttat
     2221 cggagttgca gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc tcaacagtat
     2281 gggcatttcg cagcctaccg tggtgttcgt ttccaaaaag gggttgcaaa aaattttgaa
     2341 cgtgcaaaaa aagctcccaa tcatccaaaa aattattatc atggattcta aaacggatta
     2401 ccagggattt cagtcgatgt acacgttcgt cacatctcat ctacctcccg gttttaatga
     2461 atacgatttt gtgccagagt ccttcgatag ggacaagaca attgcactga tcatgaactc
     2521 ctctggatct actggtctgc ctaaaggtgt cgctctgcct catagaactg cctgcgtgag
     2581 attctcgcat gccagagatc ctatttttgg caatcaaatc attccggata ctgcgatttt
     2641 aagtgttgtt ccattccatc acggttttgg aatgtttact acactcggat atttgatatg
     2701 tggatttcga gtcgtcttaa tgtatagatt tgaagaagag ctgtttctga ggagccttca
     2761 ggattacaag attcaaagtg cgctgctggt gccaacccta ttctccttct tcgccaaaag
     2821 cactctgatt gacaaatacg atttatctaa tttacacgaa attgcttctg gtggcgctcc
     2881 cctctctaag gaagtcgggg aagcggttgc caagaggttc catctgccag gtatcaggca
     2941 aggatatggg ctcactgaga ctacatcagc tattctgatt acacccgagg gggatgataa
     3001 accgggcgcg gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg atctggatac
     3061 cgggaaaacg ctgggcgtta atcaaagagg cgaactgtgt gtgagaggtc ctatgattat
     3121 gtccggttat gtaaacaatc cggaagcgac caacgccttg attgacaagg atggatggct
     3181 acattctgga gacatagctt actgggacga agacgaacac ttcttcatcg ttgaccgcct
     3241 gaagtctctg attaagtaca aaggctatca ggtggctccc gctgaattgg aatccatctt
     3301 gctccaacac cccaacatct tcgacgcagg tgtcgcaggt cttcccgacg atgacgccgg
     3361 tgaacttccc gccgccgttg ttgttttgga gcacggaaag acgatgacgg aaaaagagat
     3421 cgtggattac gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt
     3481 tgtggacgaa gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat
     3541 cctcataaag gccaagaagg gcggaaagat cgccgtgtaa ttctagagtc ggggcggccg
     3601 gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     3661 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     3721 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     3781 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga
     3841 taaggatccg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg
     3901 cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag
     3961 gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4021 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4081 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4141 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4201 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4261 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4321 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4381 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4441 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4501 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4561 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4621 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4681 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4741 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4801 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4861 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4921 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4981 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5041 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5101 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5161 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5221 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5281 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5341 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5401 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5461 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5521 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5581 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5641 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5701 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5761 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5821 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5881 ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg
     5941 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     6001 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     6061 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     6121 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     6181 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     6241 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     6301 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     6361 acaatttgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     6421 tcttcgctat tacgccagcc caagctacca tgataagtaa gtaatattaa ggtacgggag
     6481 gtacttggag cggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt
     6541 tgtgtgaatc gatagtacta acatacgctc tccatcaaaa caaaacgaaa caaaacaaac
     6601 tagcaaaata ggctgtcccc agtgcaagtg caggtgccag aacatttctc tatcgat
//
