LOCUS       sequence-378792-        4634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid that encodes for Hydra Sp5, lacking the SP box and DNA 
            binding domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4634)
  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 4634)
  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: HySp5-227
FEATURES             Location/Qualifiers
     source          1..4634
                     /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             748..774
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    807..941
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(831..850)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     885..904
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1054..1074)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1054..1072)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1093..1109)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1093..1109)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1106..1128)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1117..1133
                     /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(1141..1147,1148..1165,1166..1171))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1186..1207
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1324..1341)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1495..2083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1575..1594)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2254..3045,3046..3114))
                     /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     2877..2896
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3115..3219)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3245..3700)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3382..3403)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3594..3613
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
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 cagaacaaga agcaggagaa tacatttgtg
      661 aaggataccg tgatcgaagt gatcaaggac aacgtggatg agaattgtga cgagagcgtg
      721 atggagctgg aggtgaacgt ggagaattac ccctatgacg tgcctgatta tgcctaatct
      781 agaactatag tgagtcgtat tacgtagatc cagacatgat aagatacatt gatgagtttg
      841 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
      901 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
      961 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaattcgcg gccgcggcgc
     1021 caatgcattg ggcccggtac ccagcttttg ttccctttag tgagggttaa ttgcgcgctt
     1081 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     1141 caacatacga gccgggagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     1201 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     1261 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     1321 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     1381 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     1441 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1501 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1561 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1621 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1681 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1741 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1801 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1861 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1921 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1981 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2041 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2101 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2161 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2221 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2281 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     2341 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     2401 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     2461 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     2521 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     2581 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     2641 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     2701 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     2761 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     2821 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     2881 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     2941 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3001 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3061 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3121 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3181 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     3241 acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     3301 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     3361 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     3421 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     3481 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     3541 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     3601 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     3661 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg
     3721 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gtcgaccgcc
     3781 aattcaatat ggcgtatatg gactcatgcc aattcaatat ggtggatctg gacctgtgcc
     3841 aattcaatat ggcgtatatg gactcgtgcc aattcaatat ggtggatctg gaccccagcc
     3901 aattcaatat ggcggacttg gcaccatgcc aataatggcg gccatattgg ctatatgcca
     3961 ggatcaatat ataggcaata tccaatatgg ccctatgcca atatggctat tggccaggtt
     4021 caatactatg tattggccct atgccatata gtattccata tatgggtttt cctattgacg
     4081 tagatagccc ctcccaatgg gcggtcccat ataccatata tggggcttcc taataccgcc
     4141 catagccact cccccattga cgtcaatggt ctctatatat ggtctttcct attgacgtca
     4201 tatgggcggt cctattgacg tatatggcgc ctcccccatt gacgtcaatt acggtaaatg
     4261 gcccgcctgg ctcaatgccc attgacgtca ataggaccac ccaccattga cgtcaatggg
     4321 atggctcatt gcccattcat atccgttctc acgcccccta ttgacgtcaa tgacggtaaa
     4381 tggcccactt ggcagtacat caatatctat taatagtaac ttggcaagta cattactatt
     4441 ggaagtacgc cagggtacat tggcagtact cccattgacg tcaatggcgg taaatggccc
     4501 gcgatggctg ccaagtacat ccccattgac gtcaatgggg aggggcaatg acgcaaatgg
     4561 gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg ggaggtctat ataagcaatg
     4621 ctcgtttagg gaac
//
