LOCUS       sequence-191876-        3628 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dsRNA and riboprobe synthesis for Schmidtea mediterranea Smad2/3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3628)
  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 3628)
  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: pRRG223-smad2/3
FEATURES             Location/Qualifiers
     source          1..3628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..595
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     496..515
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     749..766
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(778..825)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        839..857
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        861..879
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     861..878
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(1284..1302)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(1306..1324)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1338..1385
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             1792..2586
                     /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(1846..1865)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2456..2475
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2604..2672,2673..3464)
                     /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(2822..2841)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
       61 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
      121 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
      181 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
      241 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
      301 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
      361 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
      421 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
      481 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
      541 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
      601 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
      661 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
      721 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cacgcgtcaa
      781 aaaacccctc aagacccgtt tagaggcccc aaggggttat gctaggcgcg ccggatccta
      841 atacgactca ctatagggag atttaggtga cactatagaa gtgaccttca tggaaacaag
      901 cattgtggaa aacactctca atgaagaggt tacatctgtt tcctatcagg agccccaata
      961 ttggtgcagt attcgatatt atgagctatc ggtggctttt ggtgaaactt ttcattgctc
     1021 gacatcatgt cttaccgtag atggatttac cgatcccgcc caatctgata gattctgtct
     1081 tggtttagtt agtaatgtca atcgtaatcc acaaacggag ttagctcgaa gacatattgg
     1141 taaaggtctc aggttgtatt atattggggg agaagtattt gctgaatgcc taagtgaaag
     1201 tgcgatcttt gtacaatccc ctaattgtaa ttatctcaat ggctggcacc ctgctacagt
     1261 ttgcaagatc ccaccaaggt ctccctttag tgagggttaa ttctccctat agtgagtcgt
     1321 attaggatcc ggcgcgccta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     1381 ttttgacgcg tcgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     1441 cgcgaatttt aacaaaattc agggcgcaag ggctgctaaa ggaagcggaa cacgtagaaa
     1501 gccagtccgc agaaacggtg ctgaccccgg atgaatgtca gctactgggc tatctggaca
     1561 agggaaaacg caagcgcaaa gagaaagcag gtagcttgca gtgggcttac atggcgatag
     1621 ctagactggg cggttttatg gacagcaagc gaaccggaat tgccagctgg ggcgccctct
     1681 ggtaaggttg ggaagccctg caaagtaaac tggatggctt tcttgccgcc aatgatctga
     1741 tggcgcaggg gatcaagatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     1801 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     1861 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     1921 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     1981 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     2041 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     2101 tcatcccacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     2161 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     2221 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     2281 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     2341 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     2401 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     2461 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     2521 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     2581 ttctgaattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     2641 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     2701 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     2761 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     2821 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     2881 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     2941 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3001 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     3061 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3121 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     3181 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     3241 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3301 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     3361 cctcccgtat cgtagttatc tacacgacgg ggagccaggc aactatggat gaacgaaata
     3421 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     3481 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     3541 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3601 cgtcagaccc cgtagaaaag atcaaagg
//
