LOCUS       sequence-378761-        7876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to test the Hydra Zic4 promoter activity (partial deletion 
            D1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7876)
  AUTHORS   Vogg MC, Ferenc J, Buzgariu WC, Perruchoud C , Papasaikas P, Sanchez
            PGL, Nuninger C , Delucinge-Vivier C , Rampon C , Beccari L, Vriz S 
            , Vincent S , Galliot B , Tsiairis CD
  TITLE     The transcription factor Zic4 acts as a transdifferentiation switch
  JOURNAL   bioRxiv 2021.12.22.47383
REFERENCE   2  (bases 1 to 7876)
  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: "bioRxiv 
            2021.12.22.47383"
COMMENT     Sequence Label: Zic4-3505-D1:Luc
FEATURES             Location/Qualifiers
     source          1..7876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     31..50
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      3229..3238
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3235..4887
                     /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(3301..3319)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    4928..5049
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4939..4958)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4993..5012
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5297..5314)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5468..6056)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5548..5567)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6227..7018,7019..7087))
                     /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     6850..6869
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7088..7192)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7219..7674
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7306..7325)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7516..7537
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    7805..7853
                     /note="synthetic polyadenylation signal"
     misc_feature    join(7867..7876,1..82)
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
ORIGIN
        1 ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt
       61 gcaggtgcca gaacatttct ctatcgatag gtacccgttg agattggtac aataaaacat
      121 gaaaaaggca aatttccgat taacaaagct ttggattttg ataaagaaaa aaaaaatcca
      181 aatttattat ttcacatgaa aggtgcgaaa gattagcgga gttttgtttt gtttttgaaa
      241 taatacaatt acaaattctt tttagaaaat gcggattaga atttattaaa atgacattaa
      301 cactgttttg agtctaagaa acgtaaaata tgctagttat ttttacaaca agcaggtaaa
      361 acaataaaaa tggcctttaa agtaattttg agtttatttc attgttgtta gctcaatcga
      421 tactagttca gtaaaatacg tttatttaaa gtattttact aaaattatgc tctttaatat
      481 tcgaaaaaag aaaacaaact ttttccaaaa atctaacatt tttcattact caaaaaaaaa
      541 ctttcttttt gcgctttacg cagcgctaca aaaaatatgt cgctaagtac taaaattaaa
      601 aaaaacaaaa aaacaacgta attacataac atgacgtaat gcaactttaa tacgcatttg
      661 cagtgaatac tttattattt atatatatat atatatactc acacatacgt atacatatat
      721 atgtataatc aaatacgtaa atagatatag atataatata tgaatatatt ataagtttaa
      781 tttattctat tgttattgat ttttaaaaga aagtcagaaa atcttaaact gaaagcaaaa
      841 aaaaaacttt ttcaataata tacttttatc attattttgt tgttgttgtt atgattgtta
      901 ttataattat aataataact tttattattg ttattattat tattattgta atcgtttatg
      961 aaaacttgat ggttgctaaa gatttaaggt aaataaatcg ccccctctcg ctattgattt
     1021 gtttatttta aataacgggc attggcgtga aattaatggc agcttatcat acgaattgct
     1081 atacattcaa agaatttttg ttagtaccca tgagtttatt ttgaggtgac ccatgctgtt
     1141 ctcattaatc taaaaccatt ggttttttga ttaaaaggat catcgctcaa ataaagtgtt
     1201 ttactcaggc gtgtgataac tgtattacta agacttcctc gatataaact tgcatcaaat
     1261 tatttctgaa taatgattat tgtcaaaaac aaagacacag ttgactaaat tcctaactta
     1321 ctttgtatta ttttttgtct gtaattttat agaccgatat gttacttttg ctacatgtgt
     1381 tagaagttgt ttgagaattc aaactgagtg aaaaaaatgg acaaataaaa caaaataaaa
     1441 cgcgtatgtc ctgctacctt tttatagctg ttttaaataa agtttctttt ggtattctaa
     1501 tgttgtgact tctaaaaaaa ttatgcaatc aaagattatc gacgcaacga taagtcaatt
     1561 aaaggaaagg ggtgctacat atgagacaac acatgtgtgt tttacatttt taaccggttt
     1621 aacgtcactc cgaacaaagt ttaaattaat gcgaacgttt tatttttgta gctaaacatc
     1681 tgagttaaaa atgcttgcgc acgttctttt aacaaaaaca gctcatctta ttgttttact
     1741 ttttattttt tattaaatta aaaaaaatat aattaaaaaa tatttttgta agaatcttaa
     1801 cagatatttt acgataaaat ttcttatcta aagtatcgtt ttcaggttac tcgcgtagac
     1861 ttataatttt caataaagat tacccaattt gtctaattag cataacaatt aattataagt
     1921 aaataaccgg agataataag taaataacct gagataaaaa cctcaaagag tacagcagat
     1981 tggaaatcat tttttttttt ttcaagaaac cgcgcgcatt cactcgtata ccttaattgt
     2041 ataatctttt agttaaacaa gcaccaaaac cggaacaaag gtagtagtat acaatcaaag
     2101 caagctttga tatctctaat atttttttca attttttttt tatccactta tttgtctgag
     2161 ctatttcttt accttaatta cgctataaaa ttttcttctc gacaagcggc gtagctttga
     2221 gtttatagct cttttctgaa gtctaattcc ttctgttttt attacaatcc tgactaagct
     2281 tcctcaaagt caaggaaaac aaatgcagtt aaaaagaaaa taatccttgt aaacacgttt
     2341 catccttcat aaaaaacgta cgctaataaa gcttttttgt ttacagttaa gcaagctaat
     2401 ttagaatttc agaattactc tttgcaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2461 aaaaaaaaaa gtaataataa taattagcca cgtcattttg caaccagatt aaaatccagg
     2521 aaatctcaat gaaattattt gggaggaagt tgtatatttc taaaatttag cagaacttat
     2581 attagcgtat atcttcgaca cacataaact tactgtgata ttacaaaatg actcaattga
     2641 gttagaaata aaatcaaata aataaactaa attttcccaa tactacaaag caggatctga
     2701 aaggttcttt taaggattct cgcttcatac tttttttgtt ttgtattatt ttgctcccgt
     2761 taattaggaa ataaactacc atgcagatta attatgggta taattgaaag ttcgtgcagt
     2821 tttattcttc gtcaaattaa atcaataatt tagttctatt tcgttttatt actataaatc
     2881 ctctaaagaa catttttgct tgttttcttt ttaaagatct cattttaatt tttaataccc
     2941 cgctatgatg tgattggatg ggatcaaatg tgattggtta aaaaagtttg aataagtatc
     3001 aagaactgtc aatcaaaata aatgagcggc caatcaggtt agttataaat ccttattagc
     3061 aacataatcc ctacacatag acagatgtac catacataat ggaaaggtaa aattgatttg
     3121 tgcccaaaag catttaaaag tctcaattta catccgaatg acaccgtatt tcagtcacct
     3181 cgagatctgc gatctaagta agcttggcat tccggtactg ttggtaaagc caccatggaa
     3241 gacgccaaaa acataaagaa aggcccggcg ccattctatc cgctggaaga tggaaccgct
     3301 ggagagcaac tgcataaggc tatgaagaga tacgccctgg ttcctggaac aattgctttt
     3361 acagatgcac atatcgaggt ggacatcact tacgctgagt acttcgaaat gtccgttcgg
     3421 ttggcagaag ctatgaaacg atatgggctg aatacaaatc acagaatcgt cgtatgcagt
     3481 gaaaactctc ttcaattctt tatgccggtg ttgggcgcgt tatttatcgg agttgcagtt
     3541 gcgcccgcga acgacattta taatgaacgt gaattgctca acagtatggg catttcgcag
     3601 cctaccgtgg tgttcgtttc caaaaagggg ttgcaaaaaa ttttgaacgt gcaaaaaaag
     3661 ctcccaatca tccaaaaaat tattatcatg gattctaaaa cggattacca gggatttcag
     3721 tcgatgtaca cgttcgtcac atctcatcta cctcccggtt ttaatgaata cgattttgtg
     3781 ccagagtcct tcgataggga caagacaatt gcactgatca tgaactcctc tggatctact
     3841 ggtctgccta aaggtgtcgc tctgcctcat agaactgcct gcgtgagatt ctcgcatgcc
     3901 agagatccta tttttggcaa tcaaatcatt ccggatactg cgattttaag tgttgttcca
     3961 ttccatcacg gttttggaat gtttactaca ctcggatatt tgatatgtgg atttcgagtc
     4021 gtcttaatgt atagatttga agaagagctg tttctgagga gccttcagga ttacaagatt
     4081 caaagtgcgc tgctggtgcc aaccctattc tccttcttcg ccaaaagcac tctgattgac
     4141 aaatacgatt tatctaattt acacgaaatt gcttctggtg gcgctcccct ctctaaggaa
     4201 gtcggggaag cggttgccaa gaggttccat ctgccaggta tcaggcaagg atatgggctc
     4261 actgagacta catcagctat tctgattaca cccgaggggg atgataaacc gggcgcggtc
     4321 ggtaaagttg ttccattttt tgaagcgaag gttgtggatc tggataccgg gaaaacgctg
     4381 ggcgttaatc aaagaggcga actgtgtgtg agaggtccta tgattatgtc cggttatgta
     4441 aacaatccgg aagcgaccaa cgccttgatt gacaaggatg gatggctaca ttctggagac
     4501 atagcttact gggacgaaga cgaacacttc ttcatcgttg accgcctgaa gtctctgatt
     4561 aagtacaaag gctatcaggt ggctcccgct gaattggaat ccatcttgct ccaacacccc
     4621 aacatcttcg acgcaggtgt cgcaggtctt cccgacgatg acgccggtga acttcccgcc
     4681 gccgttgttg ttttggagca cggaaagacg atgacggaaa aagagatcgt ggattacgtc
     4741 gccagtcaag taacaaccgc gaaaaagttg cgcggaggag ttgtgtttgt ggacgaagta
     4801 ccgaaaggtc ttaccggaaa actcgacgca agaaaaatca gagagatcct cataaaggcc
     4861 aagaagggcg gaaagatcgc cgtgtaattc tagagtcggg gcggccggcc gcttcgagca
     4921 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     4981 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     5041 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     5101 gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatccgtcg
     5161 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
     5221 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc
     5281 agcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5341 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5401 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5461 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5521 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5581 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5641 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5701 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5761 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5821 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5881 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5941 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6001 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6061 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6121 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6181 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6241 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6301 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6361 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6421 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6481 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6541 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6601 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6661 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6721 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6781 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6841 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6901 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6961 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7021 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7081 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7141 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7201 cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     7261 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     7321 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     7381 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     7441 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     7501 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     7561 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     7621 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttgccatt
     7681 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     7741 gccagcccaa gctaccatga taagtaagta atattaaggt acgggaggta cttggagcgg
     7801 ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat
     7861 agtactaaca tacgct
//
