LOCUS       sequence-192136-        3649 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dsRNA and riboprobe synthesis for Schmidtea mediterranea Arrowhead.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3649)
  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 3649)
  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: pRRG142-Arrowhead
FEATURES             Location/Qualifiers
     source          1..3649
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      8..596
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     497..516
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     750..767
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(779..826)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        840..858
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        862..880
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     862..879
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(1306..1324)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(1328..1346)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1360..1407
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             1814..2608
                     /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(1868..1887)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2478..2497
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2626..2694,2695..3486)
                     /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(2844..2863)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
       61 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
      121 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
      181 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
      241 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
      301 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
      361 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
      421 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
      481 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
      541 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
      601 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
      661 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
      721 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcacgcgtca
      781 aaaaacccct caagacccgt ttagaggccc caaggggtta tgctaggcgc gccggatcct
      841 aatacgactc actataggga gatttaggtg acactataga agtgaccttt tttcagtttt
      901 cccgttacaa atttctaaaa atgctcatga caaaacaaac gatcttcgat taagctacat
      961 ttttctcctt tagtaagttg cctcgtacaa atacagcaag tgaagcatga tatatgaaat
     1021 ggaaaatcat tcaatctatg aacccatgca tctccagaaa ccaaactgtt acatctttga
     1081 caatatgtca ttcttttttt acaatcttta caaatcactt tcccttttct tatccaacac
     1141 tgttcatctg attctgtcaa aagtgaagaa cacttgcaac aaattaagca atgtaaatga
     1201 aatctttctt taaatgtttc tttattaact tgctttaggt gataaaatga ttcctttaag
     1261 agtagaaagt cattacaacc aaaacacttt ccatcaaaag gtctcccttt agtgagggtt
     1321 aattctccct atagtgagtc gtattaggat ccggcgcgcc tagcataacc ccttggggcc
     1381 tctaaacggg tcttgagggg ttttttgacg cgtcgatttc ggcctattgg ttaaaaaatg
     1441 agctgattta acaaaaattt aacgcgaatt ttaacaaaat tcagggcgca agggctgcta
     1501 aaggaagcgg aacacgtaga aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt
     1561 cagctactgg gctatctgga caagggaaaa cgcaagcgca aagagaaagc aggtagcttg
     1621 cagtgggctt acatggcgat agctagactg ggcggtttta tggacagcaa gcgaaccgga
     1681 attgccagct ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa actggatggc
     1741 tttcttgccg ccaatgatct gatggcgcag gggatcaaga tctgatcaag agacaggatg
     1801 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     1861 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     1921 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     1981 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     2041 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     2101 agtgccgggg caggatctcc tgtcatccca ccttgctcct gccgagaaag tatccatcat
     2161 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     2221 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     2281 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     2341 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     2401 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     2461 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     2521 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     2581 ctatcgcctt cttgacgagt tcttctgaat tgaaaaagga agagtatgag tattcaacat
     2641 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     2701 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     2761 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     2821 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     2881 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     2941 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3001 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     3061 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     3121 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     3181 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     3241 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     3301 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     3361 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagccag
     3421 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     3481 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3541 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3601 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaag
//
