LOCUS       sequence-191886-        3973 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dsRNA and riboprobe synthesis for Schmidtea mediterranea eyes absent
            (eya).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3973)
  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 3973)
  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: pRRG272-eya
FEATURES             Location/Qualifiers
     source          1..3973
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      18..606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     507..526
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     760..777
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(789..836)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        850..868
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        872..890
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     872..889
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(1640..1658)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(1662..1680)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1694..1741
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             2148..2942
                     /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(2202..2221)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2812..2831
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2960..3028,3029..3820)
                     /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(3178..3197)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
       61 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
      121 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
      181 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
      241 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
      301 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
      361 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
      421 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
      481 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
      541 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
      601 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
      661 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
      721 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
      781 gcacgcgtca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctaggcgc
      841 gccggatcct aatacgactc actataggga gatttaggtg acactataga agtgacctta
      901 taaatttgcc tccggctctt ccgtcgactg caggcaacgc atctgtgaca gaaactgcca
      961 tctctctaaa agcgtcggaa tctcctggcc tttcgaaaga cgactcagaa agtcaatcta
     1021 aatcaccgca aagttgcaat ggcaattcgg aaacgaacaa caatccgggc ccgaaaatcc
     1081 agtcagacat tgccaatgaa aacgaactgg aaagaatttt tgtttgggat ttggacgaga
     1141 ctatcataat ttttcattct ctccttacag gaatttacgc tcagaaatac caaaaagatg
     1201 cttcaactgc agtcgctctc ggtttacgga tggaggaact tattttcagt ttagctgaca
     1261 atcacatgtt tttcaatgat ttagaagaat gcgatcaagt acacattgat gatgtgtcat
     1321 cagatgataa tggacaaaac ttaagcaatt acaattttct tgtcgacgga tttgctgggc
     1381 ctgctagtat ttgcaatgga gcatctacag tgatccaagg tggaccactg ggacttcctg
     1441 gaaacaccag tgtgcgcgga ggagttgatt ggatgaggaa attagcgttc cgttatcgac
     1501 gaataaaaga actgtacaat tcttacagaa acaatgtagg aggtttatta ggcacacaaa
     1561 aacgggaaca ctggctacaa ttaaggcaag acattgaaac tcttacagac aattggctta
     1621 cgttggcttt gaaggtctcc ctttagtgag ggttaattct ccctatagtg agtcgtatta
     1681 ggatccggcg cgcctagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     1741 gacgcgtcga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     1801 aattttaaca aaattcaggg cgcaagggct gctaaaggaa gcggaacacg tagaaagcca
     1861 gtccgcagaa acggtgctga ccccggatga atgtcagcta ctgggctatc tggacaaggg
     1921 aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag
     1981 actgggcggt tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta
     2041 aggttgggaa gccctgcaaa gtaaactgga tggctttctt gccgccaatg atctgatggc
     2101 gcaggggatc aagatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     2161 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     2221 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     2281 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     2341 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     2401 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     2461 cccaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     2521 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     2581 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     2641 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     2701 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     2761 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     2821 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     2881 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     2941 gaattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     3001 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     3061 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3121 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     3181 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     3241 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3301 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3361 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3421 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3481 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3541 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3601 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3661 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3721 ccgtatcgta gttatctaca cgacggggag ccaggcaact atggatgaac gaaatagaca
     3781 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3841 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3901 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3961 agaccccgta gaa
//
