LOCUS       sequence-378728-        5334 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid that encodes for Hydra Sp5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5334)
  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 5334)
  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: HySp5-420
FEATURES             Location/Qualifiers
     source          1..5334
                     /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             1327..1353
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1386..1520
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1410..1429)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1464..1483
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1633..1653)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1633..1651)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1672..1688)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1672..1688)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1685..1707)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1696..1712
                     /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(1720..1726,1727..1744,1745..1750))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1765..1786
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1903..1920)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2074..2662)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2154..2173)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2833..3624,3625..3693))
                     /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     3456..3475
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3694..3798)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3824..4279)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3961..3982)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4173..4192
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4350..5334
                     /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 accatgtctc cccctagcag ggtgcctacc
      121 acaatcagct ccaacttcaa gtcccagcac cactgcctga aggagcacat caagtattct
      181 cccctggccc tgctggcagc aacctgtaag aagatcggcc ggcctatctc tccactggag
      241 cagacaagcc caaagaagct gtttcagccc tggaaccaca ccatcgagag ccacaattat
      301 gagacaccca tctcccctaa ctctaaggtg cggcacttcc tggagacaaa cttcagcctg
      361 ccaccctccc ctccactgaa gtccgagatc atcaaggtgc cccctacaat cagaccaatg
      421 cccatgacaa acgtgatgca ggagaaggcc accctgaatt actctcagaa gctgagccct
      481 ccctcctgcc tggcatgtag cgccggacag aagtgcaacg gcatcaataa gatctctcct
      541 gtgctgctgt ctccacccgc aagcccaatc tcctggctgt tcccccagaa catcatccag
      601 tctcacccaa gcaaggtgtc catcaatgag caccacatca aggagtattc tgagcacagc
      661 caggccgacc ccacacggtt tgtgaactac gtgtataaga atgtggatag ctcccaggcc
      721 aagcctaacc tgatcatcag gcacgacaat atggtgtcta gcacccagtc ctacaacaat
      781 cgcatcttct cctctagccc ccacctgacc acaaccagcc acatctattc catgtctaca
      841 agcattcctg cacagagcca cgcagtggtg ccaaattctg tggcaacccg gagatgcagg
      901 cgctgcaagt gtccaaactg tatctccggc cagcagtctg agcctaataa gccaaagcag
      961 cacgtgtgcc acatccccgg ctgtggcaag gtgtacggca agacatccca cctgaaagca
     1021 cacctgagat ggcacgcagg cctgcggccc ttcgtgtgca actggctgtt ctgtaataag
     1081 agctttaccc ggtccgatga gctgcagcgg cacctgagaa cacacaccgg cgagaagaga
     1141 ttcgcctgcc aggactgtgg caagaggttt acacgctctg atcacctgag caagcacatg
     1201 aagacccacc agaacaagaa gcaggagaat acatttgtga aggataccgt gatcgaagtg
     1261 atcaaggaca acgtggatga gaattgtgac gagagcgtga tggagctgga ggtgaacgtg
     1321 gagaattacc cctatgacgt gcctgattat gcctaatcta gaactatagt gagtcgtatt
     1381 acgtagatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     1441 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     1501 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     1561 gggaggtgtg ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg gcccggtacc
     1621 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     1681 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccgggagcat
     1741 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     1801 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     1861 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     1921 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     1981 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2041 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2101 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2161 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2221 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2281 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2341 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2401 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2461 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2521 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2581 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2641 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2701 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2761 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2821 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2881 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     2941 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3001 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3061 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3121 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3181 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3241 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3301 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3361 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3421 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     3481 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3541 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     3601 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     3661 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     3721 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     3781 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
     3841 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     3901 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     3961 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     4021 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     4081 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     4141 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     4201 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     4261 gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     4321 tcggtgcggg cctcttcgct attacgccag tcgaccgcca attcaatatg gcgtatatgg
     4381 actcatgcca attcaatatg gtggatctgg acctgtgcca attcaatatg gcgtatatgg
     4441 actcgtgcca attcaatatg gtggatctgg accccagcca attcaatatg gcggacttgg
     4501 caccatgcca attcaatatg gcggacctgg cactgtgcca actggggagg ggtctacttg
     4561 gcacggtgcc aagtttgagg aggggtcttg gccctgtgcc aagtccgcca tattgaattg
     4621 gcatggtgcc aataatggcg gccatattgg ctatatgcca ggatcaatat ataggcaata
     4681 tccaatatgg ccctatgcca atatggctat tggccaggtt caatactatg tattggccct
     4741 atgccatata gtattccata tatgggtttt cctattgacg tagatagccc ctcccaatgg
     4801 gcggtcccat ataccatata tggggcttcc taataccgcc catagccact cccccattga
     4861 cgtcaatggt ctctatatat ggtctttcct attgacgtca tatgggcggt cctattgacg
     4921 tatatggcgc ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc
     4981 attgacgtca ataggaccac ccaccattga cgtcaatggg atggctcatt gcccattcat
     5041 atccgttctc acgcccccta ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat
     5101 caatatctat taatagtaac ttggcaagta cattactatt ggaagtacgc cagggtacat
     5161 tggcagtact cccattgacg tcaatggcgg taaatggccc gcgatggctg ccaagtacat
     5221 ccccattgac gtcaatgggg aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg
     5281 ggcggtaggc gtgcctaatg ggaggtctat ataagcaatg ctcgtttagg gaac
//
