LOCUS       sequence-191878-        3555 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dsRNA and riboprobe synthesis for Schmidtea mediterranea h.72.4d.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3555)
  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 3555)
  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: pRRG318-h.72.4d
FEATURES             Location/Qualifiers
     source          1..3555
                     /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(1213..1231)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(1235..1253)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1267..1314
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             1721..2515
                     /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(1775..1794)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2385..2404
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2533..2601,2602..3393)
                     /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(2751..2770)
                     /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 caatgaatca
      901 atagaatgtg gaatattcat catattctca tcagatagac tcacaccgac tgttggtagc
      961 atcggtttgg atttctcatt ttcacaccga ctataaggct tcatctgata attcggaacc
     1021 cccggagagc aaaattggtt atctggtttt gagttggatt tggaattcat cagcgattgg
     1081 tttaaaaggc gattttttaa aatttaataa atcgattaaa ttgaattgtg aaaaataaaa
     1141 tattgttgat ttttgttgag cgaaaaactg attgaaaatc aagaacaaag ttacaaatca
     1201 aaaaaaggtc tccctttagt gagggttaat tctccctata gtgagtcgta ttaggatccg
     1261 gcgcgcctag cataacccct tggggcctct aaacgggtct tgaggggttt tttgacgcgt
     1321 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     1381 acaaaattca gggcgcaagg gctgctaaag gaagcggaac acgtagaaag ccagtccgca
     1441 gaaacggtgc tgaccccgga tgaatgtcag ctactgggct atctggacaa gggaaaacgc
     1501 aagcgcaaag agaaagcagg tagcttgcag tgggcttaca tggcgatagc tagactgggc
     1561 ggttttatgg acagcaagcg aaccggaatt gccagctggg gcgccctctg gtaaggttgg
     1621 gaagccctgc aaagtaaact ggatggcttt cttgccgcca atgatctgat ggcgcagggg
     1681 atcaagatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     1741 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     1801 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     1861 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     1921 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     1981 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catcccacct
     2041 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     2101 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     2161 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     2221 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     2281 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     2341 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     2401 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     2461 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgaattga
     2521 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     2581 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     2641 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     2701 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     2761 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     2821 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     2881 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     2941 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     3001 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     3061 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     3121 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     3181 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     3241 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     3301 gtagttatct acacgacggg gagccaggca actatggatg aacgaaatag acagatcgct
     3361 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     3421 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     3481 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3541 gtagaaaaga tcaaa
//
