LOCUS       sequence-388953-        6750 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to test the Hydra Wnt3 promoter activity (partial deletion 
            of the repressor element-D3).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6750)
  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 6750)
  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-D3:Luc
FEATURES             Location/Qualifiers
     source          1..6750
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2014..2023
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2020..3672
                     /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(2086..2104)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3713..3834
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3724..3743)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3778..3797
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4082..4099)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4253..4841)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4333..4352)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5012..5803,5804..5872))
                     /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     5635..5654
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5873..5977)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6004..6459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6091..6110)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6301..6322
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6590..6638
                     /note="synthetic polyadenylation signal"
     misc_feature    6652..6743
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6692..6711
                     /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 taatcattat aaaaagttat aaatttataa
     1801 aaatttatct ctgcaaaaaa agatttaaca ttaacacaac tactacaaaa atctatctca
     1861 ggatactgca atttcacaag aaacttttca cattaatttt taaacacaat gcaaaaataa
     1921 acgcaaagaa tatgcagttt taaaacatgc atttttaaag aaactcgaga tctgcgatct
     1981 aagtaagctt ggcattccgg tactgttggt aaagccacca tggaagacgc caaaaacata
     2041 aagaaaggcc cggcgccatt ctatccgctg gaagatggaa ccgctggaga gcaactgcat
     2101 aaggctatga agagatacgc cctggttcct ggaacaattg cttttacaga tgcacatatc
     2161 gaggtggaca tcacttacgc tgagtacttc gaaatgtccg ttcggttggc agaagctatg
     2221 aaacgatatg ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa
     2281 ttctttatgc cggtgttggg cgcgttattt atcggagttg cagttgcgcc cgcgaacgac
     2341 atttataatg aacgtgaatt gctcaacagt atgggcattt cgcagcctac cgtggtgttc
     2401 gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa
     2461 aaaattatta tcatggattc taaaacggat taccagggat ttcagtcgat gtacacgttc
     2521 gtcacatctc atctacctcc cggttttaat gaatacgatt ttgtgccaga gtccttcgat
     2581 agggacaaga caattgcact gatcatgaac tcctctggat ctactggtct gcctaaaggt
     2641 gtcgctctgc ctcatagaac tgcctgcgtg agattctcgc atgccagaga tcctattttt
     2701 ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg ttccattcca tcacggtttt
     2761 ggaatgttta ctacactcgg atatttgata tgtggatttc gagtcgtctt aatgtataga
     2821 tttgaagaag agctgtttct gaggagcctt caggattaca agattcaaag tgcgctgctg
     2881 gtgccaaccc tattctcctt cttcgccaaa agcactctga ttgacaaata cgatttatct
     2941 aatttacacg aaattgcttc tggtggcgct cccctctcta aggaagtcgg ggaagcggtt
     3001 gccaagaggt tccatctgcc aggtatcagg caaggatatg ggctcactga gactacatca
     3061 gctattctga ttacacccga gggggatgat aaaccgggcg cggtcggtaa agttgttcca
     3121 ttttttgaag cgaaggttgt ggatctggat accgggaaaa cgctgggcgt taatcaaaga
     3181 ggcgaactgt gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa tccggaagcg
     3241 accaacgcct tgattgacaa ggatggatgg ctacattctg gagacatagc ttactgggac
     3301 gaagacgaac acttcttcat cgttgaccgc ctgaagtctc tgattaagta caaaggctat
     3361 caggtggctc ccgctgaatt ggaatccatc ttgctccaac accccaacat cttcgacgca
     3421 ggtgtcgcag gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg
     3481 gagcacggaa agacgatgac ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca
     3541 accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa aggtcttacc
     3601 ggaaaactcg acgcaagaaa aatcagagag atcctcataa aggccaagaa gggcggaaag
     3661 atcgccgtgt aattctagag tcggggcggc cggccgcttc gagcagacat gataagatac
     3721 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     3781 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     3841 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     3901 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtcgaccga tgcccttgag
     3961 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
     4021 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt
     4081 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4141 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4201 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4261 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4321 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4381 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4441 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4501 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4561 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4621 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4681 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4741 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4801 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4861 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4921 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4981 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5041 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5101 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5161 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5221 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5281 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5341 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5401 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5461 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5521 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5581 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5641 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5701 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5761 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5821 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5881 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5941 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6001 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     6061 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     6121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     6181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     6241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     6301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     6361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     6421 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttg ccattcgcca ttcaggctgc
     6481 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag cccaagctac
     6541 catgataagt aagtaatatt aaggtacggg aggtacttgg agcggccgca ataaaatatc
     6601 tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac taacatacgc
     6661 tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag
     6721 tgcaggtgcc agaacatttc tctatcgata
//
