LOCUS       sequence-385731-        6652 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to test the Hydra Wnt3 promoter activity (partial deletion 
            of the repressor element-D1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6652)
  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 6652)
  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-D1:Luc
FEATURES             Location/Qualifiers
     source          1..6652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1917..1926
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1923..3575
                     /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(1989..2007)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3616..3737
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3627..3646)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3681..3700
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3985..4002)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4156..4744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4236..4255)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4915..5706,5707..5775))
                     /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     5538..5557
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5776..5880)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5907..6362
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5994..6013)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6204..6225
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6493..6541
                     /note="synthetic polyadenylation signal"
     misc_feature    6555..6646
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6595..6614
                     /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 gaatttcaca agaaaagttt tcacattaat
     1801 ttttaaacac aatgcaaaaa taaacgcaaa gaatatgcag ttttaaaaca tgcattttta
     1861 aagaaactcg agatctgcga tctaagtaag cttggcattc cggtactgtt ggtaaagcca
     1921 ccatggaaga cgccaaaaac ataaagaaag gcccggcgcc attctatccg ctggaagatg
     1981 gaaccgctgg agagcaactg cataaggcta tgaagagata cgccctggtt cctggaacaa
     2041 ttgcttttac agatgcacat atcgaggtgg acatcactta cgctgagtac ttcgaaatgt
     2101 ccgttcggtt ggcagaagct atgaaacgat atgggctgaa tacaaatcac agaatcgtcg
     2161 tatgcagtga aaactctctt caattcttta tgccggtgtt gggcgcgtta tttatcggag
     2221 ttgcagttgc gcccgcgaac gacatttata atgaacgtga attgctcaac agtatgggca
     2281 tttcgcagcc taccgtggtg ttcgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc
     2341 aaaaaaagct cccaatcatc caaaaaatta ttatcatgga ttctaaaacg gattaccagg
     2401 gatttcagtc gatgtacacg ttcgtcacat ctcatctacc tcccggtttt aatgaatacg
     2461 attttgtgcc agagtccttc gatagggaca agacaattgc actgatcatg aactcctctg
     2521 gatctactgg tctgcctaaa ggtgtcgctc tgcctcatag aactgcctgc gtgagattct
     2581 cgcatgccag agatcctatt tttggcaatc aaatcattcc ggatactgcg attttaagtg
     2641 ttgttccatt ccatcacggt tttggaatgt ttactacact cggatatttg atatgtggat
     2701 ttcgagtcgt cttaatgtat agatttgaag aagagctgtt tctgaggagc cttcaggatt
     2761 acaagattca aagtgcgctg ctggtgccaa ccctattctc cttcttcgcc aaaagcactc
     2821 tgattgacaa atacgattta tctaatttac acgaaattgc ttctggtggc gctcccctct
     2881 ctaaggaagt cggggaagcg gttgccaaga ggttccatct gccaggtatc aggcaaggat
     2941 atgggctcac tgagactaca tcagctattc tgattacacc cgagggggat gataaaccgg
     3001 gcgcggtcgg taaagttgtt ccattttttg aagcgaaggt tgtggatctg gataccggga
     3061 aaacgctggg cgttaatcaa agaggcgaac tgtgtgtgag aggtcctatg attatgtccg
     3121 gttatgtaaa caatccggaa gcgaccaacg ccttgattga caaggatgga tggctacatt
     3181 ctggagacat agcttactgg gacgaagacg aacacttctt catcgttgac cgcctgaagt
     3241 ctctgattaa gtacaaaggc tatcaggtgg ctcccgctga attggaatcc atcttgctcc
     3301 aacaccccaa catcttcgac gcaggtgtcg caggtcttcc cgacgatgac gccggtgaac
     3361 ttcccgccgc cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg
     3421 attacgtcgc cagtcaagta acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg
     3481 acgaagtacc gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga gagatcctca
     3541 taaaggccaa gaagggcgga aagatcgccg tgtaattcta gagtcggggc ggccggccgc
     3601 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     3661 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     3721 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     3781 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg
     3841 atccgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     3901 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     3961 gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4021 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4081 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4141 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4201 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4261 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4321 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4381 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4441 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4501 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4561 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4621 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4681 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4741 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4801 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4861 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4921 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4981 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5041 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5101 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5161 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5221 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5281 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5341 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5401 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5461 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5521 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5581 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5641 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5701 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5761 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5821 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5881 cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg
     5941 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     6001 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     6061 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     6121 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     6181 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     6241 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     6301 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     6361 ttgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     6421 gctattacgc cagcccaagc taccatgata agtaagtaat attaaggtac gggaggtact
     6481 tggagcggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt
     6541 gaatcgatag tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca
     6601 aaataggctg tccccagtgc aagtgcaggt gccagaacat ttctctatcg at
//
