LOCUS       sequence-378773-        5212 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid that encodes for Zebrafish Sp5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5212)
  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 5212)
  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: ZfSp5a-377
FEATURES             Location/Qualifiers
     source          1..5212
                     /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"
     regulatory      95..104
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1205..1231
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1264..1398
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1288..1307)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1342..1361
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1511..1531)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1511..1529)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1550..1566)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1550..1566)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1563..1585)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1574..1590
                     /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(1598..1604,1605..1622,1623..1628))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1643..1664
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1781..1798)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1952..2540)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2032..2051)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2711..3502,3503..3571))
                     /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     3334..3353
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3572..3676)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3702..4157)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3839..3860)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4051..4070
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4228..5212
                     /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 atcccatcga tgccgccacc atggcagctg tggctgtgct
      121 gaggaacgac acactacagg cgtttctgca ggatcgcact ccgaactcct ctccggaaaa
      181 cagcaaacat tctcctctgg cacttttagc agcgacgtgt aaccggatcg gtcaccacca
      241 cggatccact cccacggact ttattcaagt tccctacgac accaccctcg gctccccgtc
      301 acggattttc cacccatgga gtaatgaggc caatcatcaa tccacgcttt ccagcaaccc
      361 ttcctttgga ctatcctcta aatctcacct gcagagctcc tacgcgtccc accatgagct
      421 cccactgacc cctccggcag atcctacata tccgtatgat ttctctccgg ttaagatgtt
      481 gccatgttca atgcaatctt tacagtccac ctgcccgccg acttatgtac ctgcggttac
      541 ctacgcagca cctgccccca tccctccagc catgcctagc tttgtcccgg gacactcagg
      601 cctggtgcat cagcagcaga gacagctatc cccaaatcca ggggaggata taccatggtg
      661 gagccttcag caagggaatc ctgtagccca ttcggtgcac ccacaccgct tccccatcca
      721 aagaggctta gtgttgggac acacagattt cgcacagtac caaactcaaa tcgcagcctt
      781 gctgcacacc aaatctccgc tggctaccgc ccggcgctgc cggagatgca gatgtccgaa
      841 ctgtcagtcg tccagctcta gtgatgagcc cggaaagaag aaacagcaca tctgccacat
      901 ccccggatgc gggaaggttt acggtaaaac atcccacctg aaggcgcacc tgcgctggca
      961 ctcaggagag agacccttcg tgtgtaactg gctcttctgc gggaagagtt tcacccggtc
     1021 agatgagtta cagcggcacc tgcggacaca caccggcgaa aaacgctttg tgtgtccgga
     1081 ctgctgtaaa aggttcatga ggagcgacca tttggccaaa cacgtcaaga cgcaccagaa
     1141 caagaagagc aaaagtcacg acaaaacctt cgattatgtg aaaagagaag atctgagacc
     1201 cgtgtaccca tacgatgttc cagattacgc ttagtctaga actatagtga gtcgtattac
     1261 gtagatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     1321 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     1381 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     1441 gaggtgtggg aggtttttta attcgcggcc gcggcgccaa tgcattgggc ccggtaccca
     1501 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     1561 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc gggagcataa
     1621 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     1681 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     1741 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     1801 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     1861 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     1921 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1981 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2041 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2101 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2161 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2221 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2281 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2341 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2401 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2461 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2521 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2581 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     2641 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     2701 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     2761 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     2821 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     2881 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     2941 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3001 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3061 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3121 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3181 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3241 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3301 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     3361 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3421 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3481 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     3541 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     3601 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     3661 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     3721 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     3781 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     3841 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     3901 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     3961 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     4021 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     4081 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     4141 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
     4201 ggtgcgggcc tcttcgctat tacgccagtc gaccgccaat tcaatatggc gtatatggac
     4261 tcatgccaat tcaatatggt ggatctggac ctgtgccaat tcaatatggc gtatatggac
     4321 tcgtgccaat tcaatatggt ggatctggac cccagccaat tcaatatggc ggacttggca
     4381 ccatgccaat tcaatatggc ggacctggca ctgtgccaac tggggagggg tctacttggc
     4441 acggtgccaa gtttgaggag gggtcttggc cctgtgccaa gtccgccata ttgaattggc
     4501 atggtgccaa taatggcggc catattggct atatgccagg atcaatatat aggcaatatc
     4561 caatatggcc ctatgccaat atggctattg gccaggttca atactatgta ttggccctat
     4621 gccatatagt attccatata tgggttttcc tattgacgta gatagcccct cccaatgggc
     4681 ggtcccatat accatatatg gggcttccta ataccgccca tagccactcc cccattgacg
     4741 tcaatggtct ctatatatgg tctttcctat tgacgtcata tgggcggtcc tattgacgta
     4801 tatggcgcct cccccattga cgtcaattac ggtaaatggc ccgcctggct caatgcccat
     4861 tgacgtcaat aggaccaccc accattgacg tcaatgggat ggctcattgc ccattcatat
     4921 ccgttctcac gccccctatt gacgtcaatg acggtaaatg gcccacttgg cagtacatca
     4981 atatctatta atagtaactt ggcaagtaca ttactattgg aagtacgcca gggtacattg
     5041 gcagtactcc cattgacgtc aatggcggta aatggcccgc gatggctgcc aagtacatcc
     5101 ccattgacgt caatggggag gggcaatgac gcaaatgggc gttccattga cgtaaatggg
     5161 cggtaggcgt gcctaatggg aggtctatat aagcaatgct cgtttaggga ac
//
