LOCUS       sequence-191874-        3694 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dsRNA and riboprobe synthesis for Schmidtea mediterranea Bmp4 (also 
            known as bmp).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3694)
  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 3694)
  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: pRRG199-bmp4
FEATURES             Location/Qualifiers
     source          1..3694
                     /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(1353..1371)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(1375..1393)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1407..1454
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             1861..2655
                     /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(1915..1934)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2525..2544
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2673..2741,2742..3533)
                     /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(2891..2910)
                     /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 tcggattttg
      901 gaggaatagg tcatctcaca caagtaccga tttgaaattt cttatacaaa aaatgaatca
      961 actttattta tactatgcaa atatcgttac attttatggt aataaagctc ctttgtctac
     1021 atataaccaa gaggaatcta caattcctaa agatgaaaat attcgtttaa agagagttaa
     1081 acgtgataaa actgattata ttcaaagttt ggaagatgac tgtcaaagat attctttaat
     1141 tgttaccttc aaagaagtag gatggagtaa atggatcata gcccctcaga attacaatgc
     1201 gtattattgc aaaggtaact gtccttaccc gttatcagat aattttaatg ccacaaacca
     1261 tgcaattatt caattattaa ttcatggtct aaaagacatt tctattccaa aaccgtgctg
     1321 tgtgccctac aatttacgcc cagaaaggtc tccctttagt gagggttaat tctccctata
     1381 gtgagtcgta ttaggatccg gcgcgcctag cataacccct tggggcctct aaacgggtct
     1441 tgaggggttt tttgacgcgt cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     1501 aaaatttaac gcgaatttta acaaaattca gggcgcaagg gctgctaaag gaagcggaac
     1561 acgtagaaag ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag ctactgggct
     1621 atctggacaa gggaaaacgc aagcgcaaag agaaagcagg tagcttgcag tgggcttaca
     1681 tggcgatagc tagactgggc ggttttatgg acagcaagcg aaccggaatt gccagctggg
     1741 gcgccctctg gtaaggttgg gaagccctgc aaagtaaact ggatggcttt cttgccgcca
     1801 atgatctgat ggcgcagggg atcaagatct gatcaagaga caggatgagg atcgtttcgc
     1861 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     1921 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     1981 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     2041 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     2101 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     2161 gatctcctgt catcccacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     2221 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     2281 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     2341 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     2401 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     2461 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     2521 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     2581 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     2641 gacgagttct tctgaattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2701 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2761 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     2821 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     2881 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     2941 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3001 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3061 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     3121 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     3181 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     3241 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     3301 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3361 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3421 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagccaggca actatggatg
     3481 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3541 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3601 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3661 tccactgagc gtcagacccc gtagaaaaga tcaa
//
