LOCUS       sequence-192153-        3770 bp    DNA     circular SYN 15-APR-2025
DEFINITION  dsRNA and riboprobe synthesis for Schmidtea mediterranea GS-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3770)
  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 3770)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pRRG370-GS1
FEATURES             Location/Qualifiers
     source          1..3770
                     /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(1426..1444)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(1448..1466)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1480..1527
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             1934..2728
                     /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(1988..2007)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2598..2617
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2746..2814,2815..3606)
                     /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(2964..2983)
                     /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 gtgaccttat catccatggt
      901 ttggcattga acaggaatat actttgcttg acattgactt gtacccgttt tcttggccga
      961 aattgggatt tcctggggcc caaggcccgt actattgcgg agttggaagc aataaagtct
     1021 ttggcagaga cataattgaa gcacattaca aagcatgttt atacgctggg gttaaaattg
     1081 ctggaactaa tgctgaggtt atgccagcac agtgggaatt ccaaattgga ccgtgtgaag
     1141 gaattaatgt gtccgatcat ttatggatgg caagatactt attacatagg gtagcagaag
     1201 attttggggt tgtagtatcg ttcgatccaa aaccgattgc tggagattgg aatggagccg
     1261 gtgcccatac taattattcc acaattcaaa tgcgaggtcc aggagggata aaatacattg
     1321 aagatgcaat tgaaaagatg tcgaaaaaac atgatcttca tattgaagca tatgacccta
     1381 aaaaaggtga agataataga catcgtttaa ccggacgaag gtctcccttt agtgagggtt
     1441 aattctccct atagtgagtc gtattaggat ccggcgcgcc tagcataacc ccttggggcc
     1501 tctaaacggg tcttgagggg ttttttgacg cgtcgatttc ggcctattgg ttaaaaaatg
     1561 agctgattta acaaaaattt aacgcgaatt ttaacaaaat tcagggcgca agggctgcta
     1621 aaggaagcgg aacacgtaga aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt
     1681 cagctactgg gctatctgga caagggaaaa cgcaagcgca aagagaaagc aggtagcttg
     1741 cagtgggctt acatggcgat agctagactg ggcggtttta tggacagcaa gcgaaccgga
     1801 attgccagct ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa actggatggc
     1861 tttcttgccg ccaatgatct gatggcgcag gggatcaaga tctgatcaag agacaggatg
     1921 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     1981 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     2041 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     2101 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     2161 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     2221 agtgccgggg caggatctcc tgtcatccca ccttgctcct gccgagaaag tatccatcat
     2281 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     2341 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     2401 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     2461 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     2521 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     2581 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     2641 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     2701 ctatcgcctt cttgacgagt tcttctgaat tgaaaaagga agagtatgag tattcaacat
     2761 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     2821 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     2881 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     2941 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     3001 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3061 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3121 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     3181 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     3241 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     3301 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     3361 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     3421 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     3481 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagccag
     3541 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     3601 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3661 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3721 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
//
