LOCUS       sequence-191871-        3652 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dsRNA and riboprobe synthesis for Schmidtea mediterranea Wnt1 (also 
            known as WntP-1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3652)
  AUTHORS   Roberts-Galbraith RH, Newmark PA
  TITLE     Follistatin antagonizes activin signaling and acts with notum to 
            direct planarian head regeneration.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Jan 22;110(4):1363-8. doi: 
            10.1073/pnas.1214053110. Epub 2013 Jan 7.
  PUBMED    23297191
REFERENCE   2  (bases 1 to 3652)
  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"; doi: 
            "10.1073/pnas.1214053110"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2013-01-22- 22"; volume: "110"; issue: "4"; pages: "1363-8"
COMMENT     Sequence Label: pRRG227-Wnt1
FEATURES             Location/Qualifiers
     source          1..3652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      9..597
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     498..517
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     751..768
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(780..827)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        841..859
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        863..881
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     863..880
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(1310..1328)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(1332..1350)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1364..1411
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             1818..2612
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1872..1891)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2482..2501
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2630..2698,2699..3490)
                     /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     complement(2848..2867)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
       61 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
      121 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
      181 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
      241 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
      301 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
      361 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
      421 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
      481 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
      541 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
      601 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
      661 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
      721 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcacgcgtc
      781 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctaggcg cgccggatcc
      841 taatacgact cactataggg agatttaggt gacactatag aagtgacctt tgtggcaaat
      901 tttggtcgta caagatccac ttgtgccctg gcatacacac ttcacgttca tacttttgac
      961 tgcaactctt ctacccgttc ccgtgttgtg aaggttcatt aatgtctttt tattccaatc
     1021 tggaatgtct ggatcaaaca tttttctcgc aaatcttcga ccgaattgta cgttgtcgtc
     1081 acatccttgc caaatccagt tggtttgaga tatccgtcct ttattattgc acggacagtg
     1141 tgataactta aacgaacaag cctcagcgac agtttgagcg acactagcac taagcatcgc
     1201 ataaatgaaa gcggtttcgg gaaatccttt gagcataatg tcaccaaaaa gtaaggcgga
     1261 tggattattc aaattcggtg tcggacagtt ccagcgatga taaggtctcc ctttagtgag
     1321 ggttaattct ccctatagtg agtcgtatta ggatccggcg cgcctagcat aaccccttgg
     1381 ggcctctaaa cgggtcttga ggggtttttt gacgcgtcga tttcggccta ttggttaaaa
     1441 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaattcaggg cgcaagggct
     1501 gctaaaggaa gcggaacacg tagaaagcca gtccgcagaa acggtgctga ccccggatga
     1561 atgtcagcta ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag
     1621 cttgcagtgg gcttacatgg cgatagctag actgggcggt tttatggaca gcaagcgaac
     1681 cggaattgcc agctggggcg ccctctggta aggttgggaa gccctgcaaa gtaaactgga
     1741 tggctttctt gccgccaatg atctgatggc gcaggggatc aagatctgat caagagacag
     1801 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     1861 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     1921 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     1981 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     2041 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     2101 gcgaagtgcc ggggcaggat ctcctgtcat cccaccttgc tcctgccgag aaagtatcca
     2161 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     2221 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     2281 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     2341 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     2401 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     2461 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     2521 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     2581 ccttctatcg ccttcttgac gagttcttct gaattgaaaa aggaagagta tgagtattca
     2641 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     2701 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     2761 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     2821 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     2881 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     2941 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     3001 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     3061 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3121 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     3181 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     3241 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     3301 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     3361 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     3421 ccaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     3481 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     3541 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     3601 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aa
//
