LOCUS       sequence-191824-        4340 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dsRNA and riboprobe synthesis for Schmidtea mediterranea Ndk (nou 
            darake).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4340)
  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 4340)
  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: pRRG304-Ndk
FEATURES             Location/Qualifiers
     source          1..4340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      16..604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     505..524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     758..775
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(787..834)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        848..866
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        870..888
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     870..887
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(2005..2023)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(2027..2045)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2059..2106
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             2513..3307
                     /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(2567..2586)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3177..3196
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3325..3393,3394..4185)
                     /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(3543..3562)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
       61 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
      121 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
      181 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
      241 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
      301 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
      361 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
      421 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
      481 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
      541 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
      601 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
      661 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
      721 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      781 acgcgtcaaa aaacccctca agacccgttt agaggcccca aggggttatg ctaggcgcgc
      841 cggatcctaa tacgactcac tatagggaga tttaggtgac actatagaag tgaccttatc
      901 gacatatgac aataccgatt gtcaaaccct tcaatcctga cattacgtta aatagcggtg
      961 aagaattgaa gttggagtgt ccgattgtga tgtcaagaga aggaaccgac gtgaattatc
     1021 catccaatct catgtatgaa tggaacataa agggtcaaac cgaagtctca gttcaaatga
     1081 atacgaaata ttctttcatt gaaaataatc ggattttgaa attaacgaag cccgtcacga
     1141 gtgaagacag tgggaaatat ttttgcaaag gggtcactgg attcggctca agagaagtgg
     1201 ccttcaatgt cgaagtcatt gattcatctc aagagaaatt gtgtgctgtg aaaactccga
     1261 aagattctaa atatgaacct ccgtgtttcg ttgatccaat aatgagaact ggtaaaataa
     1321 caattcgtag agaagtagga tcaacagtaa aattgtcgtg tgaagttaga ggtagccaac
     1381 caatgaagta tacttggttg agtggaaaga cattggcgaa ttggattctt aacacggagg
     1441 gtatcagagg tcctcattta acaataccga atttgaaaac tcactatact ggtcattata
     1501 catgtcaagt gcataatcaa gctggcggat tgaactttac gtatcattta atagtcacag
     1561 atccaccaaa taactcacca aaaatcttga cagacatttc aaattatact tttcgttcaa
     1621 aagaaaatat ttcaatcatt gtggaaatag aatgcgaatg caaagattta ataataaaat
     1681 ggttgaaaag ggttgacaga ttagatccgt caaataaaga gcattcttat ggactgagtt
     1741 taccgaatgc gatggaaagt gaaaagggag aatattacct cacaatagat accggtgtca
     1801 caacagtacg agttcagcct aacctgttta aaactgtgct caatatggaa tctgtgggaa
     1861 ctcatcacag tgggaaatac gttgtcacag ctataaacaa aattgtcaat cttaaaagct
     1921 ttgaatataa agttgtattt ataactgtag aattagataa agatataatt ggagttaagg
     1981 cctaaggatc ctcgataagg tctcccttta gtgagggtta attctcccta tagtgagtcg
     2041 tattaggatc cggcgcgcct agcataaccc cttggggcct ctaaacgggt cttgaggggt
     2101 tttttgacgc gtcgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     2161 acgcgaattt taacaaaatt cagggcgcaa gggctgctaa aggaagcgga acacgtagaa
     2221 agccagtccg cagaaacggt gctgaccccg gatgaatgtc agctactggg ctatctggac
     2281 aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc agtgggctta catggcgata
     2341 gctagactgg gcggttttat ggacagcaag cgaaccggaa ttgccagctg gggcgccctc
     2401 tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc caatgatctg
     2461 atggcgcagg ggatcaagat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     2521 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     2581 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     2641 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     2701 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     2761 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     2821 gtcatcccac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     2881 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     2941 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     3001 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     3061 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     3121 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     3181 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     3241 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     3301 cttctgaatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     3361 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     3421 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     3481 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     3541 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     3601 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     3661 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     3721 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     3781 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     3841 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     3901 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     3961 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     4021 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     4081 ccctcccgta tcgtagttat ctacacgacg gggagccagg caactatgga tgaacgaaat
     4141 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     4201 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4261 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4321 gcgtcagacc ccgtagaaaa
//
