LOCUS       sequence-191877-        3683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dsRNA and riboprobe synthesis for Schmidtea mediterranea Ras-related
            (also known as h.18.4a).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3683)
  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 3683)
  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: pRRG245-Ras-related
FEATURES             Location/Qualifiers
     source          1..3683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      11..599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     500..519
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     753..770
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(782..829)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        843..861
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        865..883
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     865..882
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(1343..1361)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(1365..1383)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1397..1444
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             1851..2645
                     /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(1905..1924)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2515..2534
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2663..2731,2732..3523)
                     /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(2881..2900)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
       61 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
      121 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
      181 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
      241 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
      301 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
      361 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
      421 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
      481 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
      541 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
      601 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
      661 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
      721 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcacgcg
      781 tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagg cgcgccggat
      841 cctaatacga ctcactatag ggagatttag gtgacactat agaagtgacc ttttgagatt
      901 acgaagtcga cgaatcttaa ttttctatca cattctaatg ccatttagaa aaagaaaaaa
      961 agtttttcat tctacctaaa gttcatagaa atactatctg aaaaatttag ttcatgaaaa
     1021 actgataaat aataaaatat tgaaatatct ttgtagaggg agggaggatg aatttcttca
     1081 ttggaatgtt ttgagtagaa aatgtttcaa aaaacaacaa aaaataattc aaattcccac
     1141 gaattacatt acgcaacact gattgcgtcg gaacgcattg tgcatagccc tagcagcgtt
     1201 cggtttatta ttattgtccc gactgtcaat tagtctcatg agctctttga aaactgtcaa
     1261 aacacgccaa ttgtattttg ttgagcattc gattaataca cagcctttcc atggcttttt
     1321 gacgatagcg cacaaaggtc tccctttagt gagggttaat tctccctata gtgagtcgta
     1381 ttaggatccg gcgcgcctag cataacccct tggggcctct aaacgggtct tgaggggttt
     1441 tttgacgcgt cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     1501 gcgaatttta acaaaattca gggcgcaagg gctgctaaag gaagcggaac acgtagaaag
     1561 ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag ctactgggct atctggacaa
     1621 gggaaaacgc aagcgcaaag agaaagcagg tagcttgcag tgggcttaca tggcgatagc
     1681 tagactgggc ggttttatgg acagcaagcg aaccggaatt gccagctggg gcgccctctg
     1741 gtaaggttgg gaagccctgc aaagtaaact ggatggcttt cttgccgcca atgatctgat
     1801 ggcgcagggg atcaagatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     1861 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     1921 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     1981 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     2041 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     2101 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     2161 catcccacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     2221 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     2281 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     2341 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     2401 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     2461 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     2521 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     2581 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     2641 tctgaattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2701 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2761 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2821 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     2881 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     2941 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3001 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3061 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3121 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3181 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     3241 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3301 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3361 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3421 ctcccgtatc gtagttatct acacgacggg gagccaggca actatggatg aacgaaatag
     3481 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3541 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3601 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3661 gtcagacccc gtagaaaaga tca
//
