LOCUS       sequence-191872-        4041 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dsRNA and riboprobe synthesis for Schmidtea mediterranea Smedwi-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4041)
  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 4041)
  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: pRRG757-smedwi-1
FEATURES             Location/Qualifiers
     source          1..4041
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      10..598
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     499..518
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     752..769
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(781..828)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        842..860
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        864..882
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     864..881
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(1700..1718)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(1722..1740)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1754..1801
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             2208..3002
                     /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(2262..2281)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2872..2891
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3020..3088,3089..3880)
                     /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(3238..3257)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
       61 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
      121 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
      181 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
      241 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
      301 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
      361 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
      421 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
      481 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
      541 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
      601 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      661 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
      721 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcacgcgt
      781 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctaggc gcgccggatc
      841 ctaatacgac tcactatagg gagatttagg tgacactata gaagtgacct tcgttcctaa
      901 aggcacagct catcgttttg gtgtactggt tgttgatcga aaccctagtc acttcaacaa
      961 ttttattgaa gatgttaaaa gagaaatagg aagattaaga atcaattaca caatggacag
     1021 tatttcttca tgtcgatcca atgaagttga agacgcttta acagatttta ttcgtgtaag
     1081 caaggtacat atggcattag tgttcattcc tgatgataaa gtttacgcaa aagtaaagaa
     1141 ttttacaatg tctacaggtc tattgactca gtgtgtgacg cagagaaacg gaagtaatag
     1201 agatgatcga cgtcgtaaaa cggttgcaga taaatcagtt atgcaaatgt tttctaaatt
     1261 aggatacgac ccatggggta ttaatcttaa aatggcaccg actatgattg taggattgga
     1321 tacttttcat agtaaaaccg gtaaaagatc tgttcaagca tccgtatttt caattagtgc
     1381 caaattttct cagtatatca gttttgtcaa ttcatctaaa ggtaaaaatg aatttcacga
     1441 aaatttgggc aaaaacttcc taacggccct tacaacattc caaaacaaat ttaacactat
     1501 gcctcttcgt ttgattattt atcgtgacgg tgttggagat tcacagttag catttaccaa
     1561 aaagttcgaa actgatgctg ttatgaaaat gatagagaaa atttacgaaa atcaaacttt
     1621 gccccaaatt atttatgttg tggttaagaa gagaattagt gttaaatttt tcaaagatgg
     1681 ggctaatcca aaaggtctcc ctttagtgag ggttaattct ccctatagtg agtcgtatta
     1741 ggatccggcg cgcctagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     1801 gacgcgtcga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     1861 aattttaaca aaattcaggg cgcaagggct gctaaaggaa gcggaacacg tagaaagcca
     1921 gtccgcagaa acggtgctga ccccggatga atgtcagcta ctgggctatc tggacaaggg
     1981 aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag
     2041 actgggcggt tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta
     2101 aggttgggaa gccctgcaaa gtaaactgga tggctttctt gccgccaatg atctgatggc
     2161 gcaggggatc aagatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     2221 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     2281 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     2341 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     2401 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     2461 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     2521 cccaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     2581 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     2641 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     2701 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     2761 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     2821 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     2881 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     2941 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     3001 gaattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     3061 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     3121 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3181 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     3241 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     3301 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3361 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3421 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3481 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3541 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3601 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3661 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3721 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3781 ccgtatcgta gttatctaca cgacggggag ccaggcaact atggatgaac gaaatagaca
     3841 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3901 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3961 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4021 agaccccgta gaaaagatca a
//
