LOCUS       sequence-191880-        3603 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dsRNA and riboprobe synthesis for Schmidtea mediterranea nemo-like 
            kinase (also known as h.109.4e).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3603)
  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 3603)
  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: pRRG319-nlk
FEATURES             Location/Qualifiers
     source          1..3603
                     /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(1264..1282)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(1286..1304)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1318..1365
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             1772..2566
                     /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(1826..1845)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2436..2455
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2584..2652,2653..3444)
                     /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(2802..2821)
                     /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 cttcaaatgg
      901 ttgttacgct ccaattcttt tgaacaataa atgcgaagat aatactttat gggagagacc
      961 tttgggatat ggagctttcg gagtagtttg gtcagttttt gatcctagaa ctcgtaaaaa
     1021 agcagcactg aaacgaatac cgaatatatt tcacaataca gttaccgcca aacgattata
     1081 ccgtgaactt ttcattcttt cgcacatcaa acatgagaat ataatttctc ttgtggatat
     1141 tgtcaaatcg gatcacagct ttattaatga cgaaatatac atattatgtg attttatgca
     1201 aactgatctc cataaaatta ttgtttctcc tcaagcgttg tcagtcgacc atgtcaaggt
     1261 ctccctttag tgagggttaa ttctccctat agtgagtcgt attaggatcc ggcgcgccta
     1321 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgacgcg tcgatttcgg
     1381 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaattc
     1441 agggcgcaag ggctgctaaa ggaagcggaa cacgtagaaa gccagtccgc agaaacggtg
     1501 ctgaccccgg atgaatgtca gctactgggc tatctggaca agggaaaacg caagcgcaaa
     1561 gagaaagcag gtagcttgca gtgggcttac atggcgatag ctagactggg cggttttatg
     1621 gacagcaagc gaaccggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg
     1681 caaagtaaac tggatggctt tcttgccgcc aatgatctga tggcgcaggg gatcaagatc
     1741 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     1801 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     1861 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     1921 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     1981 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     2041 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatcccacc ttgctcctgc
     2101 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     2161 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     2221 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     2281 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     2341 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     2401 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     2461 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     2521 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgaattg aaaaaggaag
     2581 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     2641 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     2701 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     2761 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     2821 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     2881 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2941 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     3001 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3061 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     3121 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     3181 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     3241 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3301 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     3361 tacacgacgg ggagccaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3421 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     3481 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3541 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3601 atc
//
