LOCUS       sequence-378798-        3576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to generate an ISH probe for Hydra Sp5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3576)
  AUTHORS   Vogg MC, Beccari L, Iglesias Olle L, Rampon C, Vriz S, Perruchoud C,
            Wenger Y, Galliot B
  TITLE     An evolutionarily-conserved Wnt3/beta-catenin/Sp5 feedback loop 
            restricts head organizer activity in Hydra.
  JOURNAL   Nat Commun. 2019 Jan 18;10(1):312. doi: 10.1038/s41467-018-08242-2.
  PUBMED    30659200
REFERENCE   2  (bases 1 to 3576)
  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: "Nat 
            Commun."; date: "2019-01-18"; volume: "10(1):312. doi"; pages: " 
            10.1038/s41467-018-08242-2"
COMMENT     Sequence Label: pGEM-T-Easy-HySp5-557
FEATURES             Location/Qualifiers
     source          1..3576
                     /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(701..719)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(702..719)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(737..753)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(737..753)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(750..772)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    761..777
                     /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(785..791,792..809,810..815))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    830..851
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(968..985)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1139..1727)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1219..1238)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1898..2689,2690..2758))
                     /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     2521..2540
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2759..2863)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2941..3396)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3078..3099)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3290..3309
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3522..3544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3536..3553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3537..3553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(3560..3576,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(3560..3576,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgaattg ggcccgacgt cgcatgctcc cggccgccat ggcggccgcg ggaattcgat
       61 ttaaggtgac tagttttacc ataaaccttt ccgcaccctg gaatatgaca tacgtgctgc
      121 ttgggtttat taggttcaga ttgctgtcct gatatgcagc ttggacattt acagcgacga
      181 catcttcggg ttgcaacgct gttcggtaca actgcatgtg attgcgctgg aattgatgta
      241 gacattgaat atatgtggga tgttgtagtt aaatgtggtg aagatgagaa tatacgattg
      301 ttatatgatt gtgtagagga gaccatgtta tcgtgtcgaa taattaaatt aggttttgct
      361 tgagaagagt cgacgttttt gtaaacgtag tttacaaaac gcgttggatc agcttgagaa
      421 tgttcggaat attcttttat gtggtgctcg ttaattgata ctttagaagg atgagattga
      481 ataatatttt gaggaaataa ccatgagatt ggcgaggcag gcggagaaag caaaactgga
      541 gatattttat ttattccatt acacttttga ccagctgaac atgcgagaca tggtggtgga
      601 gaaagttttt gtgagtaatt aatcactagt gaattcgcgg ccgcctgcag gtcgaccata
      661 tgggagagct cccaacgcgt tggatgcata gcttgagtat tctatagtgt cacctaaata
      721 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
      781 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
      841 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
      901 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
      961 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1021 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1081 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1141 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1201 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1261 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1321 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1381 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     1441 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     1501 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     1561 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     1621 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     1681 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     1741 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     1801 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     1861 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     1921 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     1981 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2041 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2101 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2161 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2221 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2281 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     2341 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     2401 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     2461 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     2521 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     2581 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     2641 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     2701 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     2761 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     2821 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     2881 tgccacctga tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     2941 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     3001 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     3061 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     3121 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     3181 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     3241 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3301 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3361 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat tcgccattca ggctgcgcaa
     3421 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     3481 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     3541 aacgacggcc agtgaattgt aatacgactc actata
//
