LOCUS       sequence-378762-        5340 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to test the Hydra Zic4 promoter activity (partial deletion 
            D6).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5340)
  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 5340)
  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-D6:Luc
FEATURES             Location/Qualifiers
     source          1..5340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     31..50
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      693..702
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             699..2351
                     /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(765..783)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2392..2513
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2403..2422)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2457..2476
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2761..2778)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2932..3520)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3012..3031)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3691..4482,4483..4551))
                     /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     4314..4333
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4552..4656)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4683..5138
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4770..4789)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4980..5001
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5269..5317
                     /note="synthetic polyadenylation signal"
     misc_feature    join(5331..5340,1..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 gtaccctcaa ttgagttaga aataaaatca
      121 aataaataaa ctaaattttc ccaatactac aaagcaggat ctgaaaggtt cttttaagga
      181 ttctcgcttc atactttttt tgttttgtat tattttgctc ccgttaatta ggaaataaac
      241 taccatgcag attaattatg ggtataattg aaagttcgtg cagttttatt cttcgtcaaa
      301 ttaaatcaat aatttagttc tatttcgttt tattactata aatcctctaa agaacatttt
      361 tgcttgtttt ctttttaaag atctcatttt aatttttaat accccgctat gatgtgattg
      421 gatgggatca aatgtgattg gttaaaaaag tttgaataag tatcaagaac tgtcaatcaa
      481 aataaatgag cggccaatca ggttagttat aaatccttat tagcaacata atccctacac
      541 atagacagat gtaccataca taatggaaag gtaaaattga tttgtgccca aaagcattta
      601 aaagtctcaa tttacatccg aatgacaccg tatttcagtc acctcgagat ctgcgatcta
      661 agtaagcttg gcattccggt actgttggta aagccaccat ggaagacgcc aaaaacataa
      721 agaaaggccc ggcgccattc tatccgctgg aagatggaac cgctggagag caactgcata
      781 aggctatgaa gagatacgcc ctggttcctg gaacaattgc ttttacagat gcacatatcg
      841 aggtggacat cacttacgct gagtacttcg aaatgtccgt tcggttggca gaagctatga
      901 aacgatatgg gctgaataca aatcacagaa tcgtcgtatg cagtgaaaac tctcttcaat
      961 tctttatgcc ggtgttgggc gcgttattta tcggagttgc agttgcgccc gcgaacgaca
     1021 tttataatga acgtgaattg ctcaacagta tgggcatttc gcagcctacc gtggtgttcg
     1081 tttccaaaaa ggggttgcaa aaaattttga acgtgcaaaa aaagctccca atcatccaaa
     1141 aaattattat catggattct aaaacggatt accagggatt tcagtcgatg tacacgttcg
     1201 tcacatctca tctacctccc ggttttaatg aatacgattt tgtgccagag tccttcgata
     1261 gggacaagac aattgcactg atcatgaact cctctggatc tactggtctg cctaaaggtg
     1321 tcgctctgcc tcatagaact gcctgcgtga gattctcgca tgccagagat cctatttttg
     1381 gcaatcaaat cattccggat actgcgattt taagtgttgt tccattccat cacggttttg
     1441 gaatgtttac tacactcgga tatttgatat gtggatttcg agtcgtctta atgtatagat
     1501 ttgaagaaga gctgtttctg aggagccttc aggattacaa gattcaaagt gcgctgctgg
     1561 tgccaaccct attctccttc ttcgccaaaa gcactctgat tgacaaatac gatttatcta
     1621 atttacacga aattgcttct ggtggcgctc ccctctctaa ggaagtcggg gaagcggttg
     1681 ccaagaggtt ccatctgcca ggtatcaggc aaggatatgg gctcactgag actacatcag
     1741 ctattctgat tacacccgag ggggatgata aaccgggcgc ggtcggtaaa gttgttccat
     1801 tttttgaagc gaaggttgtg gatctggata ccgggaaaac gctgggcgtt aatcaaagag
     1861 gcgaactgtg tgtgagaggt cctatgatta tgtccggtta tgtaaacaat ccggaagcga
     1921 ccaacgcctt gattgacaag gatggatggc tacattctgg agacatagct tactgggacg
     1981 aagacgaaca cttcttcatc gttgaccgcc tgaagtctct gattaagtac aaaggctatc
     2041 aggtggctcc cgctgaattg gaatccatct tgctccaaca ccccaacatc ttcgacgcag
     2101 gtgtcgcagg tcttcccgac gatgacgccg gtgaacttcc cgccgccgtt gttgttttgg
     2161 agcacggaaa gacgatgacg gaaaaagaga tcgtggatta cgtcgccagt caagtaacaa
     2221 ccgcgaaaaa gttgcgcgga ggagttgtgt ttgtggacga agtaccgaaa ggtcttaccg
     2281 gaaaactcga cgcaagaaaa atcagagaga tcctcataaa ggccaagaag ggcggaaaga
     2341 tcgccgtgta attctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca
     2401 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     2461 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     2521 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     2581 agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc gtcgaccgat gcccttgaga
     2641 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     2701 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc
     2761 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2821 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2881 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2941 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3001 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3061 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3121 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3181 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3241 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3301 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3361 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3421 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3481 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3541 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3601 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3661 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3721 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3781 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3841 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3901 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3961 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4021 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4081 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4141 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4201 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4261 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4321 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4381 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4441 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4501 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4561 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4621 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4681 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4741 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4801 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4861 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4921 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4981 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5041 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5101 taacgcgaat tttaacaaaa tattaacgct tacaatttgc cattcgccat tcaggctgcg
     5161 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc ccaagctacc
     5221 atgataagta agtaatatta aggtacggga ggtacttgga gcggccgcaa taaaatatct
     5281 ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct
//
