LOCUS       sequence-191770-        3755 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dsRNA and riboprobe synthesis for Schmidtea mediterranea 
            follistatin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3755)
  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 3755)
  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: pRRG88-follistatin
FEATURES             Location/Qualifiers
     source          1..3755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..600
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     501..520
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     754..771
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(783..830)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        844..862
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        866..884
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     866..883
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(1416..1434)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(1438..1456)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1470..1517
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             1924..2718
                     /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(1978..1997)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2588..2607
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2736..2804,2805..3596)
                     /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(2954..2973)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
       61 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
      121 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
      181 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
      241 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
      301 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
      361 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
      421 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
      481 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
      541 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
      601 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
      661 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      721 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcacgc
      781 gtcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctag gcgcgccgga
      841 tcctaatacg actcactata gggagattta ggtgacacta tagaagtgac ctttatgtcc
      901 ggtcggagag ttttgtataa tgcgagacga agacagtatc tgttcttctg aatgcatttg
      961 ttcggaggag caaaaatcaa gcggaccagt gtgcaccgca actggtcgac gctacaaaaa
     1021 taaatgcgag cttcatttgg aatcgtgttg gaccaattct cagcaaaaca ttattccatg
     1081 cgcaaaattc ggatgcgaac gacttaattg tcacaaaaga aacaaaaaat gcattttgac
     1141 tttaaatggg cagagtaaat gtgtaaataa aaaaatttgc gttggacaat tgtatgaccc
     1201 gatttgtgga gttgataatg tcacgtattc aagtcaatgt catttagagg aatctagcat
     1261 tcgccaggcg aaagaaatcc agattgccta tagtggagtt tgcagagctg acgcaaattg
     1321 tatgaatgtc gaatgtcaag aaagggacat gacgtgcatt actcaccaca gaaatggatc
     1381 accagtttgt atcaaatgcc ccacctcaag gtctcccttt agtgagggtt aattctccct
     1441 atagtgagtc gtattaggat ccggcgcgcc tagcataacc ccttggggcc tctaaacggg
     1501 tcttgagggg ttttttgacg cgtcgatttc ggcctattgg ttaaaaaatg agctgattta
     1561 acaaaaattt aacgcgaatt ttaacaaaat tcagggcgca agggctgcta aaggaagcgg
     1621 aacacgtaga aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg
     1681 gctatctgga caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt
     1741 acatggcgat agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct
     1801 ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg
     1861 ccaatgatct gatggcgcag gggatcaaga tctgatcaag agacaggatg aggatcgttt
     1921 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     1981 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     2041 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     2101 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     2161 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     2221 caggatctcc tgtcatccca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     2281 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     2341 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     2401 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     2461 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     2521 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     2581 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     2641 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     2701 cttgacgagt tcttctgaat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     2761 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     2821 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     2881 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     2941 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     3001 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3061 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3121 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3181 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3241 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     3301 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3361 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3421 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3481 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagccag gcaactatgg
     3541 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     3601 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     3661 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     3721 cgttccactg agcgtcagac cccgtagaaa agatc
//
