LOCUS       sequence-385732-        5004 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid that encodes for Hydra Sp5, lacking the SP box.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5004)
  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 5004)
  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: HySp5-310
FEATURES             Location/Qualifiers
     source          1..5004
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             997..1023
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1056..1190
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1080..1099)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1134..1153
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1303..1323)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1303..1321)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1342..1358)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1342..1358)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1355..1377)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1366..1382
                     /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(1390..1396,1397..1414,1415..1420))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1435..1456
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1573..1590)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1744..2332)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1824..1843)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2503..3294,3295..3363))
                     /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     3126..3145
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3364..3468)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3494..3949)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3631..3652)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3843..3862
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4020..5004
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccgccgcc accatgacaa acgtgatgca ggagaaggcc
      121 accctgaatt actctcagaa gctgagccct ccctcctgcc tggcatgtag cgccggacag
      181 aagtgcaacg gcatcaataa gatctctcct gtgctgctgt ctccacccgc aagcccaatc
      241 tcctggctgt tcccccagaa catcatccag tctcacccaa gcaaggtgtc catcaatgag
      301 caccacatca aggagtattc tgagcacagc caggccgacc ccacacggtt tgtgaactac
      361 gtgtataaga atgtggatag ctcccaggcc aagcctaacc tgatcatcag gcacgacaat
      421 atggtgtcta gcacccagtc ctacaacaat cgcatcttct cctctagccc ccacctgacc
      481 acaaccagcc acatctattc catgtctaca agcattcctg cacagagcca cgcagtggtg
      541 ccaaattctg tggcaacccg gagatgcagg cgctgcaagt gtccaaactg tatctccggc
      601 cagcagtctg agcctaataa gccaaagcag cacgtgtgcc acatccccgg ctgtggcaag
      661 gtgtacggca agacatccca cctgaaagca cacctgagat ggcacgcagg cctgcggccc
      721 ttcgtgtgca actggctgtt ctgtaataag agctttaccc ggtccgatga gctgcagcgg
      781 cacctgagaa cacacaccgg cgagaagaga ttcgcctgcc aggactgtgg caagaggttt
      841 acacgctctg atcacctgag caagcacatg aagacccacc agaacaagaa gcaggagaat
      901 acatttgtga aggataccgt gatcgaagtg atcaaggaca acgtggatga gaattgtgac
      961 gagagcgtga tggagctgga ggtgaacgtg gagaattacc cctatgacgt gcctgattat
     1021 gcctaatcta gaactatagt gagtcgtatt acgtagatcc agacatgata agatacattg
     1081 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     1141 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1201 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taattcgcgg
     1261 ccgcggcgcc aatgcattgg gcccggtacc cagcttttgt tccctttagt gagggttaat
     1321 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     1381 aattccacac aacatacgag ccgggagcat aaagtgtaaa gcctggggtg cctaatgagt
     1441 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     1501 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1561 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1621 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1681 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1741 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1801 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1861 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1921 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1981 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2041 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2101 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2161 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2221 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2281 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2341 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2401 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2461 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2521 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2581 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2641 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2701 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2761 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2821 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     2881 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     2941 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3001 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3061 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3121 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3181 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3241 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3301 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3361 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3421 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3481 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     3541 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     3601 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     3661 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     3721 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     3781 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     3841 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     3901 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
     3961 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     4021 tcgaccgcca attcaatatg gcgtatatgg actcatgcca attcaatatg gtggatctgg
     4081 acctgtgcca attcaatatg gcgtatatgg actcgtgcca attcaatatg gtggatctgg
     4141 accccagcca attcaatatg gcggacttgg caccatgcca attcaatatg gcggacctgg
     4201 cactgtgcca actggggagg ggtctacttg gcacggtgcc aagtttgagg aggggtcttg
     4261 gccctgtgcc aagtccgcca tattgaattg gcatggtgcc aataatggcg gccatattgg
     4321 ctatatgcca ggatcaatat ataggcaata tccaatatgg ccctatgcca atatggctat
     4381 tggccaggtt caatactatg tattggccct atgccatata gtattccata tatgggtttt
     4441 cctattgacg tagatagccc ctcccaatgg gcggtcccat ataccatata tggggcttcc
     4501 taataccgcc catagccact cccccattga cgtcaatggt ctctatatat ggtctttcct
     4561 attgacgtca tatgggcggt cctattgacg tatatggcgc ctcccccatt gacgtcaatt
     4621 acggtaaatg gcccgcctgg ctcaatgccc attgacgtca ataggaccac ccaccattga
     4681 cgtcaatggg atggctcatt gcccattcat atccgttctc acgcccccta ttgacgtcaa
     4741 tgacggtaaa tggcccactt ggcagtacat caatatctat taatagtaac ttggcaagta
     4801 cattactatt ggaagtacgc cagggtacat tggcagtact cccattgacg tcaatggcgg
     4861 taaatggccc gcgatggctg ccaagtacat ccccattgac gtcaatgggg aggggcaatg
     4921 acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg ggaggtctat
     4981 ataagcaatg ctcgtttagg gaac
//
