LOCUS       sequence-378760-        6862 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to test the Hydra Zic4 promoter activity (partial deletion 
            D3).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6862)
  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 6862)
  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-D3:Luc
FEATURES             Location/Qualifiers
     source          1..6862
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     31..50
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      2215..2224
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2221..3873
                     /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(2287..2305)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3914..4035
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3925..3944)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3979..3998
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4283..4300)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4454..5042)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4534..4553)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5213..6004,6005..6073))
                     /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     5836..5855
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6074..6178)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6205..6660
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6292..6311)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6502..6523
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6791..6839
                     /note="synthetic polyadenylation signal"
     misc_feature    join(6853..6862,1..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 gtaccatgag tttattttga ggtgacccat
      121 gctgttctca ttaatctaaa accattggtt ttttgattaa aaggatcatc gctcaaataa
      181 agtgttttac tcaggcgtgt gataactgta ttactaagac ttcctcgata taaacttgca
      241 tcaaattatt tctgaataat gattattgtc aaaaacaaag acacagttga ctaaattcct
      301 aacttacttt gtattatttt ttgtctgtaa ttttatagac cgatatgtta cttttgctac
      361 atgtgttaga agttgtttga gaattcaaac tgagtgaaaa aaatggacaa ataaaacaaa
      421 ataaaacgcg tatgtcctgc taccttttta tagctgtttt aaataaagtt tcttttggta
      481 ttctaatgtt gtgacttcta aaaaaattat gcaatcaaag attatcgacg caacgataag
      541 tcaattaaag gaaaggggtg ctacatatga gacaacacat gtgtgtttta catttttaac
      601 cggtttaacg tcactccgaa caaagtttaa attaatgcga acgttttatt tttgtagcta
      661 aacatctgag ttaaaaatgc ttgcgcacgt tcttttaaca aaaacagctc atcttattgt
      721 tttacttttt attttttatt aaattaaaaa aaatataatt aaaaaatatt tttgtaagaa
      781 tcttaacaga tattttacga taaaatttct tatctaaagt atcgttttca ggttactcgc
      841 gtagacttat aattttcaat aaagattacc caatttgtct aattagcata acaattaatt
      901 ataagtaaat aaccggagat aataagtaaa taacctgaga taaaaacctc aaagagtaca
      961 gcagattgga aatcattttt ttttttttca agaaaccgcg cgcattcact cgtatacctt
     1021 aattgtataa tcttttagtt aaacaagcac caaaaccgga acaaaggtag tagtatacaa
     1081 tcaaagcaag ctttgatatc tctaatattt ttttcaattt tttttttatc cacttatttg
     1141 tctgagctat ttctttacct taattacgct ataaaatttt cttctcgaca agcggcgtag
     1201 ctttgagttt atagctcttt tctgaagtct aattccttct gtttttatta caatcctgac
     1261 taagcttcct caaagtcaag gaaaacaaat gcagttaaaa agaaaataat ccttgtaaac
     1321 acgtttcatc cttcataaaa aacgtacgct aataaagctt ttttgtttac agttaagcaa
     1381 gctaatttag aatttcagaa ttactctttg caaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     1441 aaaaaaaaaa aaaaaagtaa taataataat tagccacgtc attttgcaac cagattaaaa
     1501 tccaggaaat ctcaatgaaa ttatttggga ggaagttgta tatttctaaa atttagcaga
     1561 acttatatta gcgtatatct tcgacacaca taaacttact gtgatattac aaaatgactc
     1621 aattgagtta gaaataaaat caaataaata aactaaattt tcccaatact acaaagcagg
     1681 atctgaaagg ttcttttaag gattctcgct tcatactttt tttgttttgt attattttgc
     1741 tcccgttaat taggaaataa actaccatgc agattaatta tgggtataat tgaaagttcg
     1801 tgcagtttta ttcttcgtca aattaaatca ataatttagt tctatttcgt tttattacta
     1861 taaatcctct aaagaacatt tttgcttgtt ttctttttaa agatctcatt ttaattttta
     1921 ataccccgct atgatgtgat tggatgggat caaatgtgat tggttaaaaa agtttgaata
     1981 agtatcaaga actgtcaatc aaaataaatg agcggccaat caggttagtt ataaatcctt
     2041 attagcaaca taatccctac acatagacag atgtaccata cataatggaa aggtaaaatt
     2101 gatttgtgcc caaaagcatt taaaagtctc aatttacatc cgaatgacac cgtatttcag
     2161 tcacctcgag atctgcgatc taagtaagct tggcattccg gtactgttgg taaagccacc
     2221 atggaagacg ccaaaaacat aaagaaaggc ccggcgccat tctatccgct ggaagatgga
     2281 accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc tggaacaatt
     2341 gcttttacag atgcacatat cgaggtggac atcacttacg ctgagtactt cgaaatgtcc
     2401 gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag aatcgtcgta
     2461 tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt tatcggagtt
     2521 gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag tatgggcatt
     2581 tcgcagccta ccgtggtgtt cgtttccaaa aaggggttgc aaaaaatttt gaacgtgcaa
     2641 aaaaagctcc caatcatcca aaaaattatt atcatggatt ctaaaacgga ttaccaggga
     2701 tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa tgaatacgat
     2761 tttgtgccag agtccttcga tagggacaag acaattgcac tgatcatgaa ctcctctgga
     2821 tctactggtc tgcctaaagg tgtcgctctg cctcatagaa ctgcctgcgt gagattctcg
     2881 catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat tttaagtgtt
     2941 gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat atgtggattt
     3001 cgagtcgtct taatgtatag atttgaagaa gagctgtttc tgaggagcct tcaggattac
     3061 aagattcaaa gtgcgctgct ggtgccaacc ctattctcct tcttcgccaa aagcactctg
     3121 attgacaaat acgatttatc taatttacac gaaattgctt ctggtggcgc tcccctctct
     3181 aaggaagtcg gggaagcggt tgccaagagg ttccatctgc caggtatcag gcaaggatat
     3241 gggctcactg agactacatc agctattctg attacacccg agggggatga taaaccgggc
     3301 gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga taccgggaaa
     3361 acgctgggcg ttaatcaaag aggcgaactg tgtgtgagag gtcctatgat tatgtccggt
     3421 tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg gctacattct
     3481 ggagacatag cttactggga cgaagacgaa cacttcttca tcgttgaccg cctgaagtct
     3541 ctgattaagt acaaaggcta tcaggtggct cccgctgaat tggaatccat cttgctccaa
     3601 caccccaaca tcttcgacgc aggtgtcgca ggtcttcccg acgatgacgc cggtgaactt
     3661 cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga gatcgtggat
     3721 tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt gtttgtggac
     3781 gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga gatcctcata
     3841 aaggccaaga agggcggaaa gatcgccgtg taattctaga gtcggggcgg ccggccgctt
     3901 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     3961 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     4021 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     4081 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat
     4141 ccgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
     4201 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
     4261 gccggcagcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4321 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4381 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4441 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4501 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4561 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4621 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4681 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4741 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4801 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4861 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4921 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4981 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5041 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5101 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5161 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5221 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5281 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5341 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5401 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5461 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5521 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5581 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5641 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5701 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5761 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5821 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5881 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5941 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6001 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6061 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6121 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6181 catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     6241 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     6301 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     6361 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     6421 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     6481 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     6541 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     6601 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     6661 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     6721 tattacgcca gcccaagcta ccatgataag taagtaatat taaggtacgg gaggtacttg
     6781 gagcggccgc aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga
     6841 atcgatagta ctaacatacg ct
//
