LOCUS       sequence-378759-        7370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to test the Hydra Zic4 promoter activity (partial deletion 
            D2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7370)
  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 7370)
  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-D2:Luc
FEATURES             Location/Qualifiers
     source          1..7370
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     31..50
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      2723..2732
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2729..4381
                     /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(2795..2813)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    4422..4543
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4433..4452)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4487..4506
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4791..4808)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4962..5550)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5042..5061)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5721..6512,6513..6581))
                     /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     6344..6363
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6582..6686)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6713..7168
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6800..6819)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7010..7031
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    7299..7347
                     /note="synthetic polyadenylation signal"
     misc_feature    join(7361..7370,1..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 gtaccaaaac aaaaaaacaa cgtaattaca
      121 taacatgacg taatgcaact ttaatacgca tttgcagtga atactttatt atttatatat
      181 atatatatat actcacacat acgtatacat atatatgtat aatcaaatac gtaaatagat
      241 atagatataa tatatgaata tattataagt ttaatttatt ctattgttat tgatttttaa
      301 aagaaagtca gaaaatctta aactgaaagc aaaaaaaaaa ctttttcaat aatatacttt
      361 tatcattatt ttgttgttgt tgttatgatt gttattataa ttataataat aacttttatt
      421 attgttatta ttattattat tgtaatcgtt tatgaaaact tgatggttgc taaagattta
      481 aggtaaataa atcgccccct ctcgctattg atttgtttat tttaaataac gggcattggc
      541 gtgaaattaa tggcagctta tcatacgaat tgctatacat tcaaagaatt tttgttagta
      601 cccatgagtt tattttgagg tgacccatgc tgttctcatt aatctaaaac cattggtttt
      661 ttgattaaaa ggatcatcgc tcaaataaag tgttttactc aggcgtgtga taactgtatt
      721 actaagactt cctcgatata aacttgcatc aaattatttc tgaataatga ttattgtcaa
      781 aaacaaagac acagttgact aaattcctaa cttactttgt attatttttt gtctgtaatt
      841 ttatagaccg atatgttact tttgctacat gtgttagaag ttgtttgaga attcaaactg
      901 agtgaaaaaa atggacaaat aaaacaaaat aaaacgcgta tgtcctgcta cctttttata
      961 gctgttttaa ataaagtttc ttttggtatt ctaatgttgt gacttctaaa aaaattatgc
     1021 aatcaaagat tatcgacgca acgataagtc aattaaagga aaggggtgct acatatgaga
     1081 caacacatgt gtgttttaca tttttaaccg gtttaacgtc actccgaaca aagtttaaat
     1141 taatgcgaac gttttatttt tgtagctaaa catctgagtt aaaaatgctt gcgcacgttc
     1201 ttttaacaaa aacagctcat cttattgttt tactttttat tttttattaa attaaaaaaa
     1261 atataattaa aaaatatttt tgtaagaatc ttaacagata ttttacgata aaatttctta
     1321 tctaaagtat cgttttcagg ttactcgcgt agacttataa ttttcaataa agattaccca
     1381 atttgtctaa ttagcataac aattaattat aagtaaataa ccggagataa taagtaaata
     1441 acctgagata aaaacctcaa agagtacagc agattggaaa tcattttttt ttttttcaag
     1501 aaaccgcgcg cattcactcg tataccttaa ttgtataatc ttttagttaa acaagcacca
     1561 aaaccggaac aaaggtagta gtatacaatc aaagcaagct ttgatatctc taatattttt
     1621 ttcaattttt tttttatcca cttatttgtc tgagctattt ctttacctta attacgctat
     1681 aaaattttct tctcgacaag cggcgtagct ttgagtttat agctcttttc tgaagtctaa
     1741 ttccttctgt ttttattaca atcctgacta agcttcctca aagtcaagga aaacaaatgc
     1801 agttaaaaag aaaataatcc ttgtaaacac gtttcatcct tcataaaaaa cgtacgctaa
     1861 taaagctttt ttgtttacag ttaagcaagc taatttagaa tttcagaatt actctttgca
     1921 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaagtaata ataataatta
     1981 gccacgtcat tttgcaacca gattaaaatc caggaaatct caatgaaatt atttgggagg
     2041 aagttgtata tttctaaaat ttagcagaac ttatattagc gtatatcttc gacacacata
     2101 aacttactgt gatattacaa aatgactcaa ttgagttaga aataaaatca aataaataaa
     2161 ctaaattttc ccaatactac aaagcaggat ctgaaaggtt cttttaagga ttctcgcttc
     2221 atactttttt tgttttgtat tattttgctc ccgttaatta ggaaataaac taccatgcag
     2281 attaattatg ggtataattg aaagttcgtg cagttttatt cttcgtcaaa ttaaatcaat
     2341 aatttagttc tatttcgttt tattactata aatcctctaa agaacatttt tgcttgtttt
     2401 ctttttaaag atctcatttt aatttttaat accccgctat gatgtgattg gatgggatca
     2461 aatgtgattg gttaaaaaag tttgaataag tatcaagaac tgtcaatcaa aataaatgag
     2521 cggccaatca ggttagttat aaatccttat tagcaacata atccctacac atagacagat
     2581 gtaccataca taatggaaag gtaaaattga tttgtgccca aaagcattta aaagtctcaa
     2641 tttacatccg aatgacaccg tatttcagtc acctcgagat ctgcgatcta agtaagcttg
     2701 gcattccggt actgttggta aagccaccat ggaagacgcc aaaaacataa agaaaggccc
     2761 ggcgccattc tatccgctgg aagatggaac cgctggagag caactgcata aggctatgaa
     2821 gagatacgcc ctggttcctg gaacaattgc ttttacagat gcacatatcg aggtggacat
     2881 cacttacgct gagtacttcg aaatgtccgt tcggttggca gaagctatga aacgatatgg
     2941 gctgaataca aatcacagaa tcgtcgtatg cagtgaaaac tctcttcaat tctttatgcc
     3001 ggtgttgggc gcgttattta tcggagttgc agttgcgccc gcgaacgaca tttataatga
     3061 acgtgaattg ctcaacagta tgggcatttc gcagcctacc gtggtgttcg tttccaaaaa
     3121 ggggttgcaa aaaattttga acgtgcaaaa aaagctccca atcatccaaa aaattattat
     3181 catggattct aaaacggatt accagggatt tcagtcgatg tacacgttcg tcacatctca
     3241 tctacctccc ggttttaatg aatacgattt tgtgccagag tccttcgata gggacaagac
     3301 aattgcactg atcatgaact cctctggatc tactggtctg cctaaaggtg tcgctctgcc
     3361 tcatagaact gcctgcgtga gattctcgca tgccagagat cctatttttg gcaatcaaat
     3421 cattccggat actgcgattt taagtgttgt tccattccat cacggttttg gaatgtttac
     3481 tacactcgga tatttgatat gtggatttcg agtcgtctta atgtatagat ttgaagaaga
     3541 gctgtttctg aggagccttc aggattacaa gattcaaagt gcgctgctgg tgccaaccct
     3601 attctccttc ttcgccaaaa gcactctgat tgacaaatac gatttatcta atttacacga
     3661 aattgcttct ggtggcgctc ccctctctaa ggaagtcggg gaagcggttg ccaagaggtt
     3721 ccatctgcca ggtatcaggc aaggatatgg gctcactgag actacatcag ctattctgat
     3781 tacacccgag ggggatgata aaccgggcgc ggtcggtaaa gttgttccat tttttgaagc
     3841 gaaggttgtg gatctggata ccgggaaaac gctgggcgtt aatcaaagag gcgaactgtg
     3901 tgtgagaggt cctatgatta tgtccggtta tgtaaacaat ccggaagcga ccaacgcctt
     3961 gattgacaag gatggatggc tacattctgg agacatagct tactgggacg aagacgaaca
     4021 cttcttcatc gttgaccgcc tgaagtctct gattaagtac aaaggctatc aggtggctcc
     4081 cgctgaattg gaatccatct tgctccaaca ccccaacatc ttcgacgcag gtgtcgcagg
     4141 tcttcccgac gatgacgccg gtgaacttcc cgccgccgtt gttgttttgg agcacggaaa
     4201 gacgatgacg gaaaaagaga tcgtggatta cgtcgccagt caagtaacaa ccgcgaaaaa
     4261 gttgcgcgga ggagttgtgt ttgtggacga agtaccgaaa ggtcttaccg gaaaactcga
     4321 cgcaagaaaa atcagagaga tcctcataaa ggccaagaag ggcggaaaga tcgccgtgta
     4381 attctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     4441 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     4501 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     4561 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     4621 ctacaaatgt ggtaaaatcg ataaggatcc gtcgaccgat gcccttgaga gccttcaacc
     4681 cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct
     4741 tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact
     4801 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4861 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4921 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4981 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5041 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5101 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5161 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5221 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5281 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5341 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5401 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5461 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5521 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5581 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5641 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5701 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5761 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5821 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5881 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5941 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6001 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6061 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6121 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6181 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6241 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6301 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6361 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6421 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6481 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6541 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6601 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6661 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc
     6721 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     6781 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     6841 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     6901 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     6961 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     7021 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     7081 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     7141 tttaacaaaa tattaacgct tacaatttgc cattcgccat tcaggctgcg caactgttgg
     7201 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc ccaagctacc atgataagta
     7261 agtaatatta aggtacggga ggtacttgga gcggccgcaa taaaatatct ttattttcat
     7321 tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct
//
