LOCUS       sequence-378763-        6355 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to test the Hydra Zic4 promoter activity (partial deletion 
            D4).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6355)
  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 6355)
  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-D4:Luc
FEATURES             Location/Qualifiers
     source          1..6355
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     31..50
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      1708..1717
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1714..3366
                     /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(1780..1798)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3407..3528
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3418..3437)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3472..3491
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3776..3793)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3947..4535)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4027..4046)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4706..5497,5498..5566))
                     /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     5329..5348
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5567..5671)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5698..6153
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5785..5804)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5995..6016
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6284..6332
                     /note="synthetic polyadenylation signal"
     misc_feature    join(6346..6355,1..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 gtaccgttta acgtcactcc gaacaaagtt
      121 taaattaatg cgaacgtttt atttttgtag ctaaacatct gagttaaaaa tgcttgcgca
      181 cgttctttta acaaaaacag ctcatcttat tgttttactt tttatttttt attaaattaa
      241 aaaaaatata attaaaaaat atttttgtaa gaatcttaac agatatttta cgataaaatt
      301 tcttatctaa agtatcgttt tcaggttact cgcgtagact tataattttc aataaagatt
      361 acccaatttg tctaattagc ataacaatta attataagta aataaccgga gataataagt
      421 aaataacctg agataaaaac ctcaaagagt acagcagatt ggaaatcatt tttttttttt
      481 tcaagaaacc gcgcgcattc actcgtatac cttaattgta taatctttta gttaaacaag
      541 caccaaaacc ggaacaaagg tagtagtata caatcaaagc aagctttgat atctctaata
      601 tttttttcaa tttttttttt atccacttat ttgtctgagc tatttcttta ccttaattac
      661 gctataaaat tttcttctcg acaagcggcg tagctttgag tttatagctc ttttctgaag
      721 tctaattcct tctgttttta ttacaatcct gactaagctt cctcaaagtc aaggaaaaca
      781 aatgcagtta aaaagaaaat aatccttgta aacacgtttc atccttcata aaaaacgtac
      841 gctaataaag cttttttgtt tacagttaag caagctaatt tagaatttca gaattactct
      901 ttgcaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaag taataataat
      961 aattagccac gtcattttgc aaccagatta aaatccagga aatctcaatg aaattatttg
     1021 ggaggaagtt gtatatttct aaaatttagc agaacttata ttagcgtata tcttcgacac
     1081 acataaactt actgtgatat tacaaaatga ctcaattgag ttagaaataa aatcaaataa
     1141 ataaactaaa ttttcccaat actacaaagc aggatctgaa aggttctttt aaggattctc
     1201 gcttcatact ttttttgttt tgtattattt tgctcccgtt aattaggaaa taaactacca
     1261 tgcagattaa ttatgggtat aattgaaagt tcgtgcagtt ttattcttcg tcaaattaaa
     1321 tcaataattt agttctattt cgttttatta ctataaatcc tctaaagaac atttttgctt
     1381 gttttctttt taaagatctc attttaattt ttaatacccc gctatgatgt gattggatgg
     1441 gatcaaatgt gattggttaa aaaagtttga ataagtatca agaactgtca atcaaaataa
     1501 atgagcggcc aatcaggtta gttataaatc cttattagca acataatccc tacacataga
     1561 cagatgtacc atacataatg gaaaggtaaa attgatttgt gcccaaaagc atttaaaagt
     1621 ctcaatttac atccgaatga caccgtattt cagtcacctc gagatctgcg atctaagtaa
     1681 gcttggcatt ccggtactgt tggtaaagcc accatggaag acgccaaaaa cataaagaaa
     1741 ggcccggcgc cattctatcc gctggaagat ggaaccgctg gagagcaact gcataaggct
     1801 atgaagagat acgccctggt tcctggaaca attgctttta cagatgcaca tatcgaggtg
     1861 gacatcactt acgctgagta cttcgaaatg tccgttcggt tggcagaagc tatgaaacga
     1921 tatgggctga atacaaatca cagaatcgtc gtatgcagtg aaaactctct tcaattcttt
     1981 atgccggtgt tgggcgcgtt atttatcgga gttgcagttg cgcccgcgaa cgacatttat
     2041 aatgaacgtg aattgctcaa cagtatgggc atttcgcagc ctaccgtggt gttcgtttcc
     2101 aaaaaggggt tgcaaaaaat tttgaacgtg caaaaaaagc tcccaatcat ccaaaaaatt
     2161 attatcatgg attctaaaac ggattaccag ggatttcagt cgatgtacac gttcgtcaca
     2221 tctcatctac ctcccggttt taatgaatac gattttgtgc cagagtcctt cgatagggac
     2281 aagacaattg cactgatcat gaactcctct ggatctactg gtctgcctaa aggtgtcgct
     2341 ctgcctcata gaactgcctg cgtgagattc tcgcatgcca gagatcctat ttttggcaat
     2401 caaatcattc cggatactgc gattttaagt gttgttccat tccatcacgg ttttggaatg
     2461 tttactacac tcggatattt gatatgtgga tttcgagtcg tcttaatgta tagatttgaa
     2521 gaagagctgt ttctgaggag ccttcaggat tacaagattc aaagtgcgct gctggtgcca
     2581 accctattct ccttcttcgc caaaagcact ctgattgaca aatacgattt atctaattta
     2641 cacgaaattg cttctggtgg cgctcccctc tctaaggaag tcggggaagc ggttgccaag
     2701 aggttccatc tgccaggtat caggcaagga tatgggctca ctgagactac atcagctatt
     2761 ctgattacac ccgaggggga tgataaaccg ggcgcggtcg gtaaagttgt tccatttttt
     2821 gaagcgaagg ttgtggatct ggataccggg aaaacgctgg gcgttaatca aagaggcgaa
     2881 ctgtgtgtga gaggtcctat gattatgtcc ggttatgtaa acaatccgga agcgaccaac
     2941 gccttgattg acaaggatgg atggctacat tctggagaca tagcttactg ggacgaagac
     3001 gaacacttct tcatcgttga ccgcctgaag tctctgatta agtacaaagg ctatcaggtg
     3061 gctcccgctg aattggaatc catcttgctc caacacccca acatcttcga cgcaggtgtc
     3121 gcaggtcttc ccgacgatga cgccggtgaa cttcccgccg ccgttgttgt tttggagcac
     3181 ggaaagacga tgacggaaaa agagatcgtg gattacgtcg ccagtcaagt aacaaccgcg
     3241 aaaaagttgc gcggaggagt tgtgtttgtg gacgaagtac cgaaaggtct taccggaaaa
     3301 ctcgacgcaa gaaaaatcag agagatcctc ataaaggcca agaagggcgg aaagatcgcc
     3361 gtgtaattct agagtcgggg cggccggccg cttcgagcag acatgataag atacattgat
     3421 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     3481 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     3541 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     3601 aacctctaca aatgtggtaa aatcgataag gatccgtcga ccgatgccct tgagagcctt
     3661 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
     3721 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc
     3781 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3841 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3901 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3961 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4021 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4081 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4141 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4201 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4261 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4321 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4381 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4441 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4501 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4561 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4621 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4681 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4741 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4801 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4861 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4921 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4981 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5041 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5101 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5161 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5221 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5281 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5341 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5401 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5461 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5521 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5581 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5641 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     5701 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     5761 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     5821 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     5881 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     5941 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     6001 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     6061 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     6121 cgaattttaa caaaatatta acgcttacaa tttgccattc gccattcagg ctgcgcaact
     6181 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagcccaag ctaccatgat
     6241 aagtaagtaa tattaaggta cgggaggtac ttggagcggc cgcaataaaa tatctttatt
     6301 ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gtactaacat acgct
//
