LOCUS       sequence-192134-        4236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dsRNA and riboprobe synthesis for Schmidtea mediterranea F-spondin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4236)
  AUTHORS   Roberts-Galbraith RH, Brubacher JL, Newmark PA
  TITLE     A functional genomics screen in planarians reveals regulators of 
            whole-brain regeneration.
  JOURNAL   Elife. 2016 Sep 9;5. pii: e17002. doi: 10.7554/eLife.17002.
  PUBMED    27612384
REFERENCE   2  (bases 1 to 4236)
  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"; journalName: "Elife."; 
            date: "2016-09-9"; volume: "5. pii: e17002. doi"; pages: " 
            10.7554/eLife.17002"
COMMENT     Sequence Label: pRRG882-F-spondin
FEATURES             Location/Qualifiers
     source          1..4236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..595
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     496..515
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     749..766
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(778..825)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        839..857
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        861..879
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     861..878
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(1892..1910)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(1914..1932)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1946..1993
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             2400..3194
                     /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(2454..2473)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3064..3083
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3212..3280,3281..4072)
                     /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(3430..3449)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
       61 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
      121 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
      181 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
      241 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
      301 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
      361 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
      421 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
      481 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
      541 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
      601 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
      661 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
      721 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cacgcgtcaa
      781 aaaacccctc aagacccgtt tagaggcccc aaggggttat gctaggcgcg ccggatccta
      841 atacgactca ctatagggag atttaggtga cactatagaa gtgaccttaa acaaaagagc
      901 atggttcatt ggaccaatct tataggggcc acacatactg gcgattactt tatatattat
      961 aacggtggtc aagcaagcca ggcagttcaa agtatttgtt catatggaga tagcacggtt
     1021 ttgaaacagc agttcgcaga agagacaata aaaccgcatt tgaagagtta ttttaatact
     1081 tcaggcatgt ggaatgaaga cgaaattgaa cagagtagat cgggattcat atcggttaat
     1141 cgtactcatc atttctttag ctttctcaca atgatgggcc caagtcctga ttggtgtact
     1201 ggagtagcag ctgtagacat gtgtatgaat gattgtacat ggaaggatga tttcaccatg
     1261 gatctattcc catttgatgc tggaattaaa aacggtttca catatttccc agaaaatccg
     1321 gacaggcaag acattcctga tccaattaga acaatcaaca ctacatatat gacccaattc
     1381 cctttcacag caaatgttcc agttgcccgg gtaatttatc aaaagatcaa accaaaacac
     1441 aattgggagt gcaccaagat gaatgcaaaa caagtaaaag aatttgatga aatgaattca
     1501 aacaaaaacg gaaattctca attcgatgga agtagtttga cgaaaaaacg ccgacacatt
     1561 aacaagaaaa gtccccttga tgatcccact ttggccaata tggctacatt tttgtgtata
     1621 acatctccat ggagtgagtg gactaaatgt tcagttacgt gtggcattgg gacagagatg
     1681 agatctcgag acatgttgaa aaatgctaaa acagaattat gtcgtcactt acctttgctt
     1741 gaaagtcgga catgcgaagg aagaaaacgt tcgtgtgatt tctcagcact atgtagcctt
     1801 ctttcctggt caagatgggg tccatgcaat gcaacttgtg agatgcacgg ataccaacag
     1861 agatctcgaa tgtttgcaag attaaggtct ccctttagtg agggttaatt ctccctatag
     1921 tgagtcgtat taggatccgg cgcgcctagc ataacccctt ggggcctcta aacgggtctt
     1981 gaggggtttt ttgacgcgtc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     2041 aaatttaacg cgaattttaa caaaattcag ggcgcaaggg ctgctaaagg aagcggaaca
     2101 cgtagaaagc cagtccgcag aaacggtgct gaccccggat gaatgtcagc tactgggcta
     2161 tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt agcttgcagt gggcttacat
     2221 ggcgatagct agactgggcg gttttatgga cagcaagcga accggaattg ccagctgggg
     2281 cgccctctgg taaggttggg aagccctgca aagtaaactg gatggctttc ttgccgccaa
     2341 tgatctgatg gcgcagggga tcaagatctg atcaagagac aggatgagga tcgtttcgca
     2401 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     2461 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     2521 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     2581 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     2641 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     2701 atctcctgtc atcccacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     2761 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     2821 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     2881 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     2941 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     3001 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     3061 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     3121 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     3181 acgagttctt ctgaattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     3241 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     3301 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     3361 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     3421 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     3481 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     3541 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     3601 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     3661 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     3721 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     3781 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     3841 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     3901 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     3961 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agccaggcaa ctatggatga
     4021 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     4081 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     4141 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     4201 ccactgagcg tcagaccccg tagaaaagat caaagg
//
