LOCUS       sequence-378746-        3589 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to generate an ISH probe for Hydra Nematocilin A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3589)
  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 3589)
  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-NematocilinA
FEATURES             Location/Qualifiers
     source          1..3589
                     /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(714..732)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(715..732)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(750..766)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(750..766)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(763..785)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    774..790
                     /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(798..804,805..822,823..828))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    843..864
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(981..998)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1152..1740)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1232..1251)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1911..2702,2703..2771))
                     /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     2534..2553
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2772..2876)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2954..3409)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3091..3112)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3303..3322
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3535..3557
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3549..3566
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3550..3566
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(3573..3589,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(3573..3589,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgaattg ggcccgacgt cgcatgctcc cggccgccat ggcggccgcg ggaattcgat
       61 tcgagagcac gatcttgttg atgaaaaact tcaacagcaa attgaaaata ttaaaaggca
      121 aacaacatac gatttcgttc aataccaaga agcttctgag aattcttacc agttacagct
      181 acaagaacat aaaaaccgaa tggcaaaaga aacacaagca cttgcgcaac aaaaagagga
      241 aaatattcat ttaaaagcaa ttattgaaga aatgaatgca aaaatatata aactggacgg
      301 aaaggtttct tcttaccatg aacaaaatac tattctaatt cacacacttg aagtcgaacg
      361 tcgcgctgct gctgcaacat gtcacgagct tgaaaagaaa cttcaagaac tccaagaaca
      421 ttacaacacc aaagtccgtg agttaaacat tgtcagcagt gtcaacatac caatcgactt
      481 agaacttgaa tcattatcgc aattaataga agctgaagca aaacgattag atgttgcttt
      541 atctaacccc tcaagtgaac ttgtttctac tgtacgtgga gaactggtgt caaaccgaag
      601 tcactatgta cacaacgcta gtccccgtaa tagaatcact agtgaattcg cggccgcctg
      661 caggtcgacc atatgggaga gctcccaacg cgttggatgc atagcttgag tattctatag
      721 tgtcacctaa atagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
      781 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
      841 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
      901 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
      961 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     1021 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     1081 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1141 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     1201 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     1261 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1321 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     1381 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     1441 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     1501 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     1561 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     1621 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     1681 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     1741 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     1801 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     1861 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     1921 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     1981 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     2041 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     2101 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     2161 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     2221 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     2281 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     2341 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     2401 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     2461 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     2521 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     2581 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     2641 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     2701 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     2761 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     2821 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     2881 tttccccgaa aagtgccacc tgatgcggtg tgaaataccg cacagatgcg taaggagaaa
     2941 ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     3001 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     3061 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     3121 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     3181 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     3241 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     3301 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     3361 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc cattcgccat
     3421 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     3481 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     3541 cacgacgttg taaaacgacg gccagtgaat tgtaatacga ctcactata
//
