LOCUS       sequence-378744-        5085 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid that encodes for Hydra Sp5, lacking the DNA binding domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5085)
  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 5085)
  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: HySp5-337
FEATURES             Location/Qualifiers
     source          1..5085
                     /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             1078..1104
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1137..1271
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1161..1180)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1215..1234
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1384..1404)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1384..1402)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1423..1439)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1423..1439)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1436..1458)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1447..1463
                     /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(1471..1477,1478..1495,1496..1501))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1516..1537
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1654..1671)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1825..2413)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1905..1924)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2584..3375,3376..3444))
                     /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     3207..3226
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3445..3549)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3575..4030)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3712..3733)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3924..3943
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4101..5085
                     /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 cagaacaaga agcaggagaa tacatttgtg aaggataccg tgatcgaagt gatcaaggac
     1021 aacgtggatg agaattgtga cgagagcgtg atggagctgg aggtgaacgt ggagaattac
     1081 ccctatgacg tgcctgatta tgcctaatct agaactatag tgagtcgtat tacgtagatc
     1141 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     1201 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     1261 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     1321 gggaggtttt ttaattcgcg gccgcggcgc caatgcattg ggcccggtac ccagcttttg
     1381 ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt
     1441 gtgaaattgt tatccgctca caattccaca caacatacga gccgggagca taaagtgtaa
     1501 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     1561 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     1621 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     1681 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     1741 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1801 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1861 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1921 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1981 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2041 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2101 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2161 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2221 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     2281 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2341 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2401 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2461 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     2521 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     2581 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     2641 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     2701 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     2761 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     2821 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     2881 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     2941 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3001 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     3061 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     3121 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     3181 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     3241 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     3301 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     3361 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     3421 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     3481 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     3541 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
     3601 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accgataggc cgaaatcggc
     3661 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3721 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     3781 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     3841 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     3901 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     3961 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4021 cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     4081 gcctcttcgc tattacgcca gtcgaccgcc aattcaatat ggcgtatatg gactcatgcc
     4141 aattcaatat ggtggatctg gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc
     4201 aattcaatat ggtggatctg gaccccagcc aattcaatat ggcggacttg gcaccatgcc
     4261 aattcaatat ggcggacctg gcactgtgcc aactggggag gggtctactt ggcacggtgc
     4321 caagtttgag gaggggtctt ggccctgtgc caagtccgcc atattgaatt ggcatggtgc
     4381 caataatggc ggccatattg gctatatgcc aggatcaata tataggcaat atccaatatg
     4441 gccctatgcc aatatggcta ttggccaggt tcaatactat gtattggccc tatgccatat
     4501 agtattccat atatgggttt tcctattgac gtagatagcc cctcccaatg ggcggtccca
     4561 tataccatat atggggcttc ctaataccgc ccatagccac tcccccattg acgtcaatgg
     4621 tctctatata tggtctttcc tattgacgtc atatgggcgg tcctattgac gtatatggcg
     4681 cctcccccat tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc cattgacgtc
     4741 aataggacca cccaccattg acgtcaatgg gatggctcat tgcccattca tatccgttct
     4801 cacgccccct attgacgtca atgacggtaa atggcccact tggcagtaca tcaatatcta
     4861 ttaatagtaa cttggcaagt acattactat tggaagtacg ccagggtaca ttggcagtac
     4921 tcccattgac gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca tccccattga
     4981 cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat gggcggtagg
     5041 cgtgcctaat gggaggtcta tataagcaat gctcgtttag ggaac
//
