LOCUS       sequence-378764-        5848 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to test the Hydra Zic4 promoter activity (partial deletion 
            D5).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5848)
  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 5848)
  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-D5:Luc
FEATURES             Location/Qualifiers
     source          1..5848
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     31..50
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      1201..1210
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1207..2859
                     /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(1273..1291)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2900..3021
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2911..2930)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2965..2984
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3269..3286)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3440..4028)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3520..3539)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4199..4990,4991..5059))
                     /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     4822..4841
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5060..5164)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5191..5646
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5278..5297)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5488..5509
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5777..5825
                     /note="synthetic polyadenylation signal"
     misc_feature    join(5839..5848,1..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 gtaccttttt caattttttt tttatccact
      121 tatttgtctg agctatttct ttaccttaat tacgctataa aattttcttc tcgacaagcg
      181 gcgtagcttt gagtttatag ctcttttctg aagtctaatt ccttctgttt ttattacaat
      241 cctgactaag cttcctcaaa gtcaaggaaa acaaatgcag ttaaaaagaa aataatcctt
      301 gtaaacacgt ttcatccttc ataaaaaacg tacgctaata aagctttttt gtttacagtt
      361 aagcaagcta atttagaatt tcagaattac tctttgcaaa aaaaaaaaaa aaaaaaaaaa
      421 aaaaaaaaaa aaaaaaaaaa aagtaataat aataattagc cacgtcattt tgcaaccaga
      481 ttaaaatcca ggaaatctca atgaaattat ttgggaggaa gttgtatatt tctaaaattt
      541 agcagaactt atattagcgt atatcttcga cacacataaa cttactgtga tattacaaaa
      601 tgactcaatt gagttagaaa taaaatcaaa taaataaact aaattttccc aatactacaa
      661 agcaggatct gaaaggttct tttaaggatt ctcgcttcat actttttttg ttttgtatta
      721 ttttgctccc gttaattagg aaataaacta ccatgcagat taattatggg tataattgaa
      781 agttcgtgca gttttattct tcgtcaaatt aaatcaataa tttagttcta tttcgtttta
      841 ttactataaa tcctctaaag aacatttttg cttgttttct ttttaaagat ctcattttaa
      901 tttttaatac cccgctatga tgtgattgga tgggatcaaa tgtgattggt taaaaaagtt
      961 tgaataagta tcaagaactg tcaatcaaaa taaatgagcg gccaatcagg ttagttataa
     1021 atccttatta gcaacataat ccctacacat agacagatgt accatacata atggaaaggt
     1081 aaaattgatt tgtgcccaaa agcatttaaa agtctcaatt tacatccgaa tgacaccgta
     1141 tttcagtcac ctcgagatct gcgatctaag taagcttggc attccggtac tgttggtaaa
     1201 gccaccatgg aagacgccaa aaacataaag aaaggcccgg cgccattcta tccgctggaa
     1261 gatggaaccg ctggagagca actgcataag gctatgaaga gatacgccct ggttcctgga
     1321 acaattgctt ttacagatgc acatatcgag gtggacatca cttacgctga gtacttcgaa
     1381 atgtccgttc ggttggcaga agctatgaaa cgatatgggc tgaatacaaa tcacagaatc
     1441 gtcgtatgca gtgaaaactc tcttcaattc tttatgccgg tgttgggcgc gttatttatc
     1501 ggagttgcag ttgcgcccgc gaacgacatt tataatgaac gtgaattgct caacagtatg
     1561 ggcatttcgc agcctaccgt ggtgttcgtt tccaaaaagg ggttgcaaaa aattttgaac
     1621 gtgcaaaaaa agctcccaat catccaaaaa attattatca tggattctaa aacggattac
     1681 cagggatttc agtcgatgta cacgttcgtc acatctcatc tacctcccgg ttttaatgaa
     1741 tacgattttg tgccagagtc cttcgatagg gacaagacaa ttgcactgat catgaactcc
     1801 tctggatcta ctggtctgcc taaaggtgtc gctctgcctc atagaactgc ctgcgtgaga
     1861 ttctcgcatg ccagagatcc tatttttggc aatcaaatca ttccggatac tgcgatttta
     1921 agtgttgttc cattccatca cggttttgga atgtttacta cactcggata tttgatatgt
     1981 ggatttcgag tcgtcttaat gtatagattt gaagaagagc tgtttctgag gagccttcag
     2041 gattacaaga ttcaaagtgc gctgctggtg ccaaccctat tctccttctt cgccaaaagc
     2101 actctgattg acaaatacga tttatctaat ttacacgaaa ttgcttctgg tggcgctccc
     2161 ctctctaagg aagtcgggga agcggttgcc aagaggttcc atctgccagg tatcaggcaa
     2221 ggatatgggc tcactgagac tacatcagct attctgatta cacccgaggg ggatgataaa
     2281 ccgggcgcgg tcggtaaagt tgttccattt tttgaagcga aggttgtgga tctggatacc
     2341 gggaaaacgc tgggcgttaa tcaaagaggc gaactgtgtg tgagaggtcc tatgattatg
     2401 tccggttatg taaacaatcc ggaagcgacc aacgccttga ttgacaagga tggatggcta
     2461 cattctggag acatagctta ctgggacgaa gacgaacact tcttcatcgt tgaccgcctg
     2521 aagtctctga ttaagtacaa aggctatcag gtggctcccg ctgaattgga atccatcttg
     2581 ctccaacacc ccaacatctt cgacgcaggt gtcgcaggtc ttcccgacga tgacgccggt
     2641 gaacttcccg ccgccgttgt tgttttggag cacggaaaga cgatgacgga aaaagagatc
     2701 gtggattacg tcgccagtca agtaacaacc gcgaaaaagt tgcgcggagg agttgtgttt
     2761 gtggacgaag taccgaaagg tcttaccgga aaactcgacg caagaaaaat cagagagatc
     2821 ctcataaagg ccaagaaggg cggaaagatc gccgtgtaat tctagagtcg gggcggccgg
     2881 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     2941 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     3001 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     3061 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat
     3121 aaggatccgt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc
     3181 gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg
     3241 acaggtgccg gcagcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3301 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3361 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3421 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3481 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3541 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3601 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3661 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3721 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3781 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3841 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3901 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3961 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4021 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4081 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4141 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4201 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4261 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4321 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4381 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4441 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4501 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4561 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4621 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4681 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4741 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4801 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4861 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4921 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4981 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5041 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5101 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5161 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
     5221 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     5281 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     5341 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     5401 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     5461 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     5521 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     5581 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     5641 caatttgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     5701 cttcgctatt acgccagccc aagctaccat gataagtaag taatattaag gtacgggagg
     5761 tacttggagc ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     5821 gtgtgaatcg atagtactaa catacgct
//
