LOCUS       sequence-378729-        6942 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to test the Hydra Wnt3 promoter activity .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6942)
  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 6942)
  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:Luc
FEATURES             Location/Qualifiers
     source          1..6942
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2206..2215
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2212..3864
                     /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(2278..2296)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3905..4026
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3916..3935)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3970..3989
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4274..4291)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4445..5033)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4525..4544)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5204..5995,5996..6064))
                     /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     5827..5846
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6065..6169)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6196..6651
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6283..6302)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6493..6514
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6782..6830
                     /note="synthetic polyadenylation signal"
     misc_feature    6844..6935
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6884..6903
                     /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 ggatactgtc aaaaagttaa acaggtcatt aaagttaact tttgtcaaag cggtatccct
     1921 attccaaaag tttcaacgta accgtgtcaa ttctttacat agcaaataga aagggtcacc
     1981 gattaaagaa aataataaaa ggattcacac gtgctaatcg tgcgctttaa agatgactga
     2041 ttgtttctaa ctttatttga aatttcacaa gaaaagtttt cacattaatt tttaaacaca
     2101 atgcaaaaat aaacgcaaag aatatgcagt tttaaaacat gcatttttaa agaaactcga
     2161 gatctgcgat ctaagtaagc ttggcattcc ggtactgttg gtaaagccac catggaagac
     2221 gccaaaaaca taaagaaagg cccggcgcca ttctatccgc tggaagatgg aaccgctgga
     2281 gagcaactgc ataaggctat gaagagatac gccctggttc ctggaacaat tgcttttaca
     2341 gatgcacata tcgaggtgga catcacttac gctgagtact tcgaaatgtc cgttcggttg
     2401 gcagaagcta tgaaacgata tgggctgaat acaaatcaca gaatcgtcgt atgcagtgaa
     2461 aactctcttc aattctttat gccggtgttg ggcgcgttat ttatcggagt tgcagttgcg
     2521 cccgcgaacg acatttataa tgaacgtgaa ttgctcaaca gtatgggcat ttcgcagcct
     2581 accgtggtgt tcgtttccaa aaaggggttg caaaaaattt tgaacgtgca aaaaaagctc
     2641 ccaatcatcc aaaaaattat tatcatggat tctaaaacgg attaccaggg atttcagtcg
     2701 atgtacacgt tcgtcacatc tcatctacct cccggtttta atgaatacga ttttgtgcca
     2761 gagtccttcg atagggacaa gacaattgca ctgatcatga actcctctgg atctactggt
     2821 ctgcctaaag gtgtcgctct gcctcataga actgcctgcg tgagattctc gcatgccaga
     2881 gatcctattt ttggcaatca aatcattccg gatactgcga ttttaagtgt tgttccattc
     2941 catcacggtt ttggaatgtt tactacactc ggatatttga tatgtggatt tcgagtcgtc
     3001 ttaatgtata gatttgaaga agagctgttt ctgaggagcc ttcaggatta caagattcaa
     3061 agtgcgctgc tggtgccaac cctattctcc ttcttcgcca aaagcactct gattgacaaa
     3121 tacgatttat ctaatttaca cgaaattgct tctggtggcg ctcccctctc taaggaagtc
     3181 ggggaagcgg ttgccaagag gttccatctg ccaggtatca ggcaaggata tgggctcact
     3241 gagactacat cagctattct gattacaccc gagggggatg ataaaccggg cgcggtcggt
     3301 aaagttgttc cattttttga agcgaaggtt gtggatctgg ataccgggaa aacgctgggc
     3361 gttaatcaaa gaggcgaact gtgtgtgaga ggtcctatga ttatgtccgg ttatgtaaac
     3421 aatccggaag cgaccaacgc cttgattgac aaggatggat ggctacattc tggagacata
     3481 gcttactggg acgaagacga acacttcttc atcgttgacc gcctgaagtc tctgattaag
     3541 tacaaaggct atcaggtggc tcccgctgaa ttggaatcca tcttgctcca acaccccaac
     3601 atcttcgacg caggtgtcgc aggtcttccc gacgatgacg ccggtgaact tcccgccgcc
     3661 gttgttgttt tggagcacgg aaagacgatg acggaaaaag agatcgtgga ttacgtcgcc
     3721 agtcaagtaa caaccgcgaa aaagttgcgc ggaggagttg tgtttgtgga cgaagtaccg
     3781 aaaggtctta ccggaaaact cgacgcaaga aaaatcagag agatcctcat aaaggccaag
     3841 aagggcggaa agatcgccgt gtaattctag agtcggggcg gccggccgct tcgagcagac
     3901 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     3961 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     4021 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     4081 gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tccgtcgacc
     4141 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat
     4201 cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc
     4261 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4321 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4381 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4441 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4501 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4561 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4621 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4681 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4741 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4801 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4861 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4921 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4981 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5041 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5101 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5161 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5221 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5281 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5341 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5401 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5461 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5521 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5581 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5641 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5701 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5761 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5821 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5881 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5941 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6001 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6061 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6121 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6181 gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     6241 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     6301 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     6361 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     6421 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     6481 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     6541 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     6601 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt tgccattcgc
     6661 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     6721 agcccaagct accatgataa gtaagtaata ttaaggtacg ggaggtactt ggagcggccg
     6781 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagt
     6841 actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt
     6901 ccccagtgca agtgcaggtg ccagaacatt tctctatcga ta
//
