LOCUS       sequence-378775-        5177 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid that encodes for Zebrafish Sp5-like.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5177)
  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 5177)
  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: ZfSp5l1
FEATURES             Location/Qualifiers
     source          1..5177
                     /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      91..100
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             403..432
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             406..435
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1168..1194
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1229..1363
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1253..1272)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1307..1326
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1476..1496)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1476..1494)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1515..1531)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1515..1531)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1528..1550)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1539..1555
                     /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(1563..1569,1570..1587,1588..1593))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1608..1629
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1746..1763)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1917..2505)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1997..2016)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2676..3467,3468..3536))
                     /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     3299..3318
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3537..3641)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3667..4122)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3804..3825)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4016..4035
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4193..5177
                     /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 atccgcggcc gccaccatgg ctgcattaac cttatccaga
      121 gcggacaatt tcctccacaa tttcttacag gaccgcactc ccagctcatc tccagaaagt
      181 ggtccaaaca ccctctcctt tctggccacc acatgcagtc aggcttggca ggaaggctcg
      241 cagctccctt acgagggtcc ggtgggctcc gcatccagca tgttccagct ctggagcaac
      301 gacgtggccc cgaactccag cttaagcgct caccagatga ccttcaccgt gcccaaaatg
      361 cagtttccca gccacatgca acccactctg ggctcgcact cccatcacca ccatcaccac
      421 catcatcacc accacgagct tcccctcact cctccagcgg agcctccgtc tgcatattca
      481 ttcgagcttt ccccggttaa aatgcaaggg aatgcgccat actacacgca gcacaacgcc
      541 gtcagtcaaa actttccaag cttcctccaa aacccctcag gaagagctca cctgcctgga
      601 ggtcacgttg aggatggcca gcagtggtgg agtctccccc agggtaacag cgctccctcc
      661 gggcaccctt tctccttggg acggcagctg gttttgggcc atcagcctca aattgcagca
      721 cttctgcagg gaacctccaa aggtctgttg agctctactc gtcgctgtcg ccgatgcaaa
      781 tgcccaaact gccagtccac aggaaacgga ggtgctgcat tggagtttgg aaagaaaaga
      841 ctacacattt gtcacattcc agattgtggg aaggtttata aaaaaacctc tcacctcaaa
      901 gcgcacctgc gctggcatgc tggtgaacgg cccttcattt gcaactggct cttctgcggg
      961 aagagcttca cgcgttcaga tgagctgcag aggcatctcc gtacacacac cggggagaag
     1021 cgtttcggct gtcagcagtg tggaaaacgg tttatgcgga gcgatcatct ctccaagcat
     1081 gtgaagaccc atcagagccg gaagagccgg tccagtcagc ctcctcacag cgggactgat
     1141 gctctgctga gcaacatcaa gagagagtac ccctatgacg tgcctgatta tgcctaagct
     1201 ctagaactat agtgagtcgt attacgtaga tccagacatg ataagataca ttgatgagtt
     1261 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     1321 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     1381 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg cggccgcggc
     1441 gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc
     1501 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     1561 cacaacatac gagccgggag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     1621 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     1681 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     1741 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     1801 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     1861 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1921 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1981 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2041 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2101 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2161 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2221 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2281 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2341 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     2401 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     2461 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     2521 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     2581 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     2641 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     2701 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     2761 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     2821 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     2881 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     2941 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3001 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3061 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3121 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     3181 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     3241 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     3301 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     3361 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     3421 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     3481 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     3541 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     3601 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     3661 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     3721 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     3781 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     3841 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     3901 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     3961 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     4021 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     4081 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     4141 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagtcgaccg
     4201 ccaattcaat atggcgtata tggactcatg ccaattcaat atggtggatc tggacctgtg
     4261 ccaattcaat atggcgtata tggactcgtg ccaattcaat atggtggatc tggaccccag
     4321 ccaattcaat atggcggact tggcaccatg ccaattcaat atggcggacc tggcactgtg
     4381 ccaactgggg aggggtctac ttggcacggt gccaagtttg aggaggggtc ttggccctgt
     4441 gccaagtccg ccatattgaa ttggcatggt gccaataatg gcggccatat tggctatatg
     4501 ccaggatcaa tatataggca atatccaata tggccctatg ccaatatggc tattggccag
     4561 gttcaatact atgtattggc cctatgccat atagtattcc atatatgggt tttcctattg
     4621 acgtagatag cccctcccaa tgggcggtcc catataccat atatggggct tcctaatacc
     4681 gcccatagcc actcccccat tgacgtcaat ggtctctata tatggtcttt cctattgacg
     4741 tcatatgggc ggtcctattg acgtatatgg cgcctccccc attgacgtca attacggtaa
     4801 atggcccgcc tggctcaatg cccattgacg tcaataggac cacccaccat tgacgtcaat
     4861 gggatggctc attgcccatt catatccgtt ctcacgcccc ctattgacgt caatgacggt
     4921 aaatggccca cttggcagta catcaatatc tattaatagt aacttggcaa gtacattact
     4981 attggaagta cgccagggta cattggcagt actcccattg acgtcaatgg cggtaaatgg
     5041 cccgcgatgg ctgccaagta catccccatt gacgtcaatg gggaggggca atgacgcaaa
     5101 tgggcgttcc attgacgtaa atgggcggta ggcgtgccta atgggaggtc tatataagca
     5161 atgctcgttt agggaac
//
