LOCUS       sequence-378730-        7786 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to test the Hydra Sp5 promoter activity .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7786)
  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 7786)
  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: HySp5:Luc
FEATURES             Location/Qualifiers
     source          1..7786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      3050..3059
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3056..4708
                     /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(3122..3140)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    4749..4870
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4760..4779)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4814..4833
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5118..5135)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5289..5877)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5369..5388)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6048..6839,6840..6908))
                     /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     6671..6690
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6909..7013)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7040..7495
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7127..7146)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7337..7358
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    7626..7674
                     /note="synthetic polyadenylation signal"
     misc_feature    7688..7779
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     7728..7747
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccctag ttctaattta gctctattac gttcgcaaag ttgacaaagt cgcaaatttt
       61 tttctttttc aaaagacctc ccattcattt taataaagac tggttctaat tttgctttat
      121 cacgcttata aaaattgaca aagtcgcaag tactttgttt tttaaaagac ctcccattca
      181 ttgagataaa tactagttat tttcatgtat cattgaaata gtaaacaatt cattctagtt
      241 tttatttgtc tagggcagta tttcacacct tccacaagtg cgaaacgttt tatttacttg
      301 attgagtaaa ttaatttaaa ataaaataaa aaaataagaa cttaatgtga aaaaaaaaac
      361 aaaacaaaaa caataaaaaa aaaaacaatg taaaatattt ctcatagctg ttgaaatatt
      421 aaaaagacgg aaggaaaaat ataacggcaa agctaaattc ttttctgcgt attgcttttt
      481 atctctttat tgtctgtttg ttgcttttca ctgcattcta ttttgtgctt aataaatctc
      541 aatcgatttt aaggaatgac taggatgttt cattttgtat atatcaataa ctgaaatatt
      601 aaaaatctcc tcagtgcatc cgttcgttag acaattgggg gtattaactc aattatttcc
      661 tgaatataaa ctcaacaagt aaaaaagttt catccgtaag caagcaagaa taacgacact
      721 tgtttacatt taagaatttc ttaactttat gtaaaaaaca attcttagtt aaaaacgaag
      781 taaaagggtt ttaatttttt gttttttagt tgaaaacaaa ttgctaaata aaacttaatt
      841 taaaaaaaaa aaaccaaata tttaaaatga taaactggtt aaaattaata gattatacaa
      901 accattgtaa gcatttaaaa acaatttttt ttttataaaa acaacaacaa aaaaatttca
      961 cctaattact ccgcctttgt taataaaacc tccgttttac agttaaaagt aatgtatgaa
     1021 ccgtaatccc tattacaaaa gaaatggtat attgtttata aagacgtttt gatgattgta
     1081 gcttatttta tatttttgtt gcttttgttt gttttccttg ttgttgtttt taatttgatt
     1141 agttatattc ttacggcctt acgaccacgt ttgctgtttt aaatcgcgag cgttgctttg
     1201 actttacacg aagcctctaa aacaaacata aagagaattc atcgaaagta aaaaaaatca
     1261 tgctgacctt cggtgatccg taattgaaat tgatatatat tttctcccta ttttgacata
     1321 taaatggtta aagttaatct ttttattaaa ctcaatcaat ttcaacaaaa taagtgtcaa
     1381 gtttactgct tatttcaagt aacaaataag tcttattgta aataaagtta ttgtttaaaa
     1441 gggattgaaa cttcaatcat tattgtaata aagaagaatt tcatgtaaga tttgtattta
     1501 ttaaaattaa ataaactaaa tgaaacaaaa gcgctacgtc aaatttatat tttgattatt
     1561 aagaggaatt tttttaccta atgtaaaact actgtaaaaa tcgactgaat caataaggtc
     1621 agagagacta ggtcagcgag tttggatcat taaaatcgat aacaataatt aacgatatag
     1681 tttataatga taggaaactt acacttgaca tttaaatggg aagtcctgag gctataacgt
     1741 tcgtttgtcg tgggtagata agccaattga caaaaccatc atcttatatt tttatgggcg
     1801 ccaaatgttt atcatgttta atttctttta tataaatgat aaaaacattt aaccacaaat
     1861 tatttttttt atctccaaat gaaatcaaga acttttaagt cataaaaagt agcgacagcg
     1921 ccagtgataa tcatagacaa gtgtacacat tagcttatca aaaagtacgc tagagtaaag
     1981 cttattgttt tgttttggct aggatatatc cttctcgtta aaataatttg tcctactttt
     2041 atatacgata ttcatatttt taggtttctt gtttcgatat atatatatat ttcgttatgt
     2101 ttgtatgtat atatgtgttt gtttgtatgt gtatgtaaaa atatgaaata tactttttgc
     2161 aaatctttgt agaagtttaa taaataaagt atcaagttta aaaattcctt cgatattttt
     2221 aaagcttcaa tttggttggg cgtagacaca ttagtaattg cggtaaagat cagtaagaat
     2281 tctaaataga cgttaatttt aaaacctggc ctgccccttg attatttaat ttgaaatttt
     2341 taagctgtct ccatttcaac cacagtatca atgggtcggc aaaaaaagaa agattgaacg
     2401 ttttatcaat tttaccaacg aaaaacactt aacaaatatt gtagtacttt tttaagttta
     2461 aatgtttttt gtaaaactgt tatttttaaa ataaaccttt tacttctttt tttttttttt
     2521 tgaaatcgtt ttaaactgat atttaataaa agcttaaata taaatgtggt aaagttcgta
     2581 aaaccaatga ggcaggtgcc ggcatagatg aaagtgaaag aacaattttt tttattgaac
     2641 ttcacattta ctgtggattg tcggaatgtt ttactattaa gttgatttga agtcaaaaac
     2701 aaaataacaa aatcaaccaa tgaacttcct tagaaattgt ttaatcataa accaatcaaa
     2761 tagcgtttta tatcttttaa ccaataagat aacaatttta ttgttttgtc ggcatcttaa
     2821 gatattaaaa gttaatatct tttccgcctt acgtattctg tttatcaccg cctcttagac
     2881 catcccattt gtacgtaaac agagaaaata tgatcgcaac gcgccatttc tcagtcagag
     2941 gcgtgacatt aaccccttat caaagaagcc gaagctattt aagataagat ggcggtttcc
     3001 tcgagatctg cgatctaagt aagcttggca ttccggtact gttggtaaag ccaccatgga
     3061 agacgccaaa aacataaaga aaggcccggc gccattctat ccgctggaag atggaaccgc
     3121 tggagagcaa ctgcataagg ctatgaagag atacgccctg gttcctggaa caattgcttt
     3181 tacagatgca catatcgagg tggacatcac ttacgctgag tacttcgaaa tgtccgttcg
     3241 gttggcagaa gctatgaaac gatatgggct gaatacaaat cacagaatcg tcgtatgcag
     3301 tgaaaactct cttcaattct ttatgccggt gttgggcgcg ttatttatcg gagttgcagt
     3361 tgcgcccgcg aacgacattt ataatgaacg tgaattgctc aacagtatgg gcatttcgca
     3421 gcctaccgtg gtgttcgttt ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa
     3481 gctcccaatc atccaaaaaa ttattatcat ggattctaaa acggattacc agggatttca
     3541 gtcgatgtac acgttcgtca catctcatct acctcccggt tttaatgaat acgattttgt
     3601 gccagagtcc ttcgataggg acaagacaat tgcactgatc atgaactcct ctggatctac
     3661 tggtctgcct aaaggtgtcg ctctgcctca tagaactgcc tgcgtgagat tctcgcatgc
     3721 cagagatcct atttttggca atcaaatcat tccggatact gcgattttaa gtgttgttcc
     3781 attccatcac ggttttggaa tgtttactac actcggatat ttgatatgtg gatttcgagt
     3841 cgtcttaatg tatagatttg aagaagagct gtttctgagg agccttcagg attacaagat
     3901 tcaaagtgcg ctgctggtgc caaccctatt ctccttcttc gccaaaagca ctctgattga
     3961 caaatacgat ttatctaatt tacacgaaat tgcttctggt ggcgctcccc tctctaagga
     4021 agtcggggaa gcggttgcca agaggttcca tctgccaggt atcaggcaag gatatgggct
     4081 cactgagact acatcagcta ttctgattac acccgagggg gatgataaac cgggcgcggt
     4141 cggtaaagtt gttccatttt ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct
     4201 gggcgttaat caaagaggcg aactgtgtgt gagaggtcct atgattatgt ccggttatgt
     4261 aaacaatccg gaagcgacca acgccttgat tgacaaggat ggatggctac attctggaga
     4321 catagcttac tgggacgaag acgaacactt cttcatcgtt gaccgcctga agtctctgat
     4381 taagtacaaa ggctatcagg tggctcccgc tgaattggaa tccatcttgc tccaacaccc
     4441 caacatcttc gacgcaggtg tcgcaggtct tcccgacgat gacgccggtg aacttcccgc
     4501 cgccgttgtt gttttggagc acggaaagac gatgacggaa aaagagatcg tggattacgt
     4561 cgccagtcaa gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt
     4621 accgaaaggt cttaccggaa aactcgacgc aagaaaaatc agagagatcc tcataaaggc
     4681 caagaagggc ggaaagatcg ccgtgtaatt ctagagtcgg ggcggccggc cgcttcgagc
     4741 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     4801 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     4861 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     4921 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatccgtc
     4981 gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga
     5041 ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg
     5101 cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5161 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5221 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5281 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5341 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5401 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5461 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5521 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5581 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5641 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5701 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5761 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5821 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5881 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5941 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6001 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6061 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6121 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6181 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6241 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6301 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6361 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6421 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6481 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6541 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6601 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6661 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6721 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6781 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6841 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6901 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6961 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7021 ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     7081 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     7141 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     7201 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     7261 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     7321 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     7381 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     7441 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttgccat
     7501 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     7561 cgccagccca agctaccatg ataagtaagt aatattaagg tacgggaggt acttggagcg
     7621 gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga
     7681 tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg
     7741 ctgtccccag tgcaagtgca ggtgccagaa catttctcta tcgata
//
