LOCUS       sequence-385729-        6746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to test the Hydra Wnt3 promoter activity (partial deletion 
            of the repressor element-D4).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6746)
  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 6746)
  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-D4:Luc
FEATURES             Location/Qualifiers
     source          1..6746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2011..2020
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2017..3669
                     /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(2083..2101)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3710..3831
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3721..3740)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3775..3794
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4079..4096)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4250..4838)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4330..4349)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5009..5800,5801..5869))
                     /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     5632..5651
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5870..5974)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6001..6456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6088..6107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6298..6319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6587..6635
                     /note="synthetic polyadenylation signal"
     misc_feature    6649..6740
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6689..6708
                     /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 aaaaagttaa acaggtcatt aaagttaact
     1801 tttgtcaaag cggtatccct attccaaaag tttcaacgta accgtgtcaa ttctttacat
     1861 agcaaataga aagggtcacc gattaaagaa aataataaaa ggattcacac gtgctaatcg
     1921 tgcgctttaa agatgactga ttgtttctaa ctttatttga ctcgagatct gcgatctaag
     1981 taagcttggc attccggtac tgttggtaaa gccaccatgg aagacgccaa aaacataaag
     2041 aaaggcccgg cgccattcta tccgctggaa gatggaaccg ctggagagca actgcataag
     2101 gctatgaaga gatacgccct ggttcctgga acaattgctt ttacagatgc acatatcgag
     2161 gtggacatca cttacgctga gtacttcgaa atgtccgttc ggttggcaga agctatgaaa
     2221 cgatatgggc tgaatacaaa tcacagaatc gtcgtatgca gtgaaaactc tcttcaattc
     2281 tttatgccgg tgttgggcgc gttatttatc ggagttgcag ttgcgcccgc gaacgacatt
     2341 tataatgaac gtgaattgct caacagtatg ggcatttcgc agcctaccgt ggtgttcgtt
     2401 tccaaaaagg ggttgcaaaa aattttgaac gtgcaaaaaa agctcccaat catccaaaaa
     2461 attattatca tggattctaa aacggattac cagggatttc agtcgatgta cacgttcgtc
     2521 acatctcatc tacctcccgg ttttaatgaa tacgattttg tgccagagtc cttcgatagg
     2581 gacaagacaa ttgcactgat catgaactcc tctggatcta ctggtctgcc taaaggtgtc
     2641 gctctgcctc atagaactgc ctgcgtgaga ttctcgcatg ccagagatcc tatttttggc
     2701 aatcaaatca ttccggatac tgcgatttta agtgttgttc cattccatca cggttttgga
     2761 atgtttacta cactcggata tttgatatgt ggatttcgag tcgtcttaat gtatagattt
     2821 gaagaagagc tgtttctgag gagccttcag gattacaaga ttcaaagtgc gctgctggtg
     2881 ccaaccctat tctccttctt cgccaaaagc actctgattg acaaatacga tttatctaat
     2941 ttacacgaaa ttgcttctgg tggcgctccc ctctctaagg aagtcgggga agcggttgcc
     3001 aagaggttcc atctgccagg tatcaggcaa ggatatgggc tcactgagac tacatcagct
     3061 attctgatta cacccgaggg ggatgataaa ccgggcgcgg tcggtaaagt tgttccattt
     3121 tttgaagcga aggttgtgga tctggatacc gggaaaacgc tgggcgttaa tcaaagaggc
     3181 gaactgtgtg tgagaggtcc tatgattatg tccggttatg taaacaatcc ggaagcgacc
     3241 aacgccttga ttgacaagga tggatggcta cattctggag acatagctta ctgggacgaa
     3301 gacgaacact tcttcatcgt tgaccgcctg aagtctctga ttaagtacaa aggctatcag
     3361 gtggctcccg ctgaattgga atccatcttg ctccaacacc ccaacatctt cgacgcaggt
     3421 gtcgcaggtc ttcccgacga tgacgccggt gaacttcccg ccgccgttgt tgttttggag
     3481 cacggaaaga cgatgacgga aaaagagatc gtggattacg tcgccagtca agtaacaacc
     3541 gcgaaaaagt tgcgcggagg agttgtgttt gtggacgaag taccgaaagg tcttaccgga
     3601 aaactcgacg caagaaaaat cagagagatc ctcataaagg ccaagaaggg cggaaagatc
     3661 gccgtgtaat tctagagtcg gggcggccgg ccgcttcgag cagacatgat aagatacatt
     3721 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     3781 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     3841 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     3901 taaaacctct acaaatgtgg taaaatcgat aaggatccgt cgaccgatgc ccttgagagc
     3961 cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat
     4021 gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct tccgcttcct
     4081 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4141 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4201 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4261 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4321 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4381 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4441 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4501 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4561 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4621 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4681 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4741 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4801 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4861 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4921 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4981 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5041 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5101 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5161 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5221 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5281 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5341 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5401 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5461 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5521 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5581 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5641 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5701 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5761 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5821 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5881 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5941 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6001 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     6061 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     6121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     6181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     6241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     6301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     6361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     6421 acgcgaattt taacaaaata ttaacgctta caatttgcca ttcgccattc aggctgcgca
     6481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagccc aagctaccat
     6541 gataagtaag taatattaag gtacgggagg tacttggagc ggccgcaata aaatatcttt
     6601 attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagtactaa catacgctct
     6661 ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc
     6721 aggtgccaga acatttctct atcgat
//
