LOCUS       sequence-378765-        4071 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to generate an ISH probe for Hydra Zic4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4071)
  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 4071)
  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: pGEM-T-Easy-Zic4
FEATURES             Location/Qualifiers
     source          1..4071
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      33..42
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      37..46
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1196..1214)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1197..1214)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1232..1248)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1232..1248)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1245..1267)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1256..1272
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1280..1286,1287..1304,1305..1310))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1325..1346
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1463..1480)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1634..2222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1714..1733)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2393..3184,3185..3253))
                     /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     3016..3035
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3254..3358)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3436..3891)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3573..3594)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3785..3804
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4017..4039
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4031..4048
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4032..4048
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(4055..4071,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(4055..4071,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgaattg ggcccgacgt cgcatgctcc cggccgccat ggcggccgcg ggaattcgat
       61 tatgccgatc acaatgtctc agatcacgtt gcttcacctc ctgggatttt taatgcacat
      121 tctgccatga tgagtccatc tcaggctgta ggaagtgtag gtacgtacgc ttcacgttca
      181 tctagtcaaa gtgaatacca acattcttct ggagtcggaa attctgattt tatgaatgga
      241 tataaaacaa tacacgagga accaccaata ttaactgcag cacagccaac tcattcaata
      301 ccaactatga tgaatgacac aaatacgcat gctgggagaa cacattgtat gtttgctagt
      361 ggtgcactgc atcataatac tgaaataaac tcacgatttc actctaattt gaatcaaaat
      421 cacggatatt cttctaaccc aagtacccct tcgtcttttt atcctctatc ccatcacgaa
      481 ttatcgcaca acagccacta tcctggaaaa tacccaaagc acgattatgc aagtgacgtt
      541 aagccagcat acaactatca gccactaaat tcatcttacg aatactacgg aagcccacag
      601 tacggatact caaatcttca acctcataac agttttcggc aacctcttaa tcaagttata
      661 tcatgcatgt ggcaagatca aaccacaaaa ggaaaaccgt gcggaaaaca gttttttgtt
      721 atgatggaca tagtgcagca tttagctgaa gatcacgtgg gcttaaatga ctcgacagaa
      781 catatatgtt attggaaaga ttgcccacga gcaggacttg ctttcaaagc taaatataaa
      841 cttattaatc acctaagagt acacacaggt gaaaaacctt ttccatgtcc attcccagga
      901 tgtggcaaac tatttgcaag atcggaaaat ttaaagatac ataagcgaac gcacaccggt
      961 gaaagaccgt ttgtgtgtga atttacaggt tgcggtagaa gatttgcaaa ctcttccgac
     1021 cgtaaaaaac acagtcacgt gcatacttcc gacaagccat atatttgtaa atacgatgga
     1081 tgcaataaaa cgtatacaca tccaagttcc cttcgaatca ctagtgaatt cgcggccgcc
     1141 tgcaggtcga ccatatggga gagctcccaa cgcgttggat gcatagcttg agtattctat
     1201 agtgtcacct aaatagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     1261 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     1321 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     1381 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1441 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1501 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     1561 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1621 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1681 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1741 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1801 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1861 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1921 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1981 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2041 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2101 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2161 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2221 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2281 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2341 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     2401 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     2461 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     2521 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     2581 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     2641 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     2701 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     2761 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     2821 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     2881 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     2941 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3001 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3061 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3121 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3181 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     3241 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     3301 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     3361 catttccccg aaaagtgcca cctgatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     3421 aaataccgca tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     3481 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     3541 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     3601 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     3661 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3721 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3781 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3841 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc
     3901 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     3961 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     4021 gtcacgacgt tgtaaaacga cggccagtga attgtaatac gactcactat a
//
