LOCUS       sequence-192155-        3663 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dsRNA and riboprobe synthesis for Schmidtea mediterranea slc1a-5 
            (also known as excitatory amino acid transporter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3663)
  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 3663)
  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: pRRG312-slc1a-5
FEATURES             Location/Qualifiers
     source          1..3663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      10..598
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     499..518
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     752..769
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(781..828)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        842..860
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        864..882
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     864..881
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(1322..1340)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(1344..1362)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1376..1423
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             1830..2624
                     /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(1884..1903)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2494..2513
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2642..2710,2711..3502)
                     /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(2860..2879)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
       61 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
      121 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
      181 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
      241 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
      301 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
      361 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
      421 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
      481 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
      541 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
      601 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      661 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
      721 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcacgcgt
      781 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctaggc gcgccggatc
      841 ctaatacgac tcactatagg gagatttagg tgacactata gaagtgacct tattaattgt
      901 ggcgcctatc ggtaaaacaa atcttgtaac tcgtttatct acaccattgt tttcctccag
      961 acatttgaat gtaataggta gagttgctga gctcgatgct gtacctaaag cagttaccca
     1021 tgcttgtaac attccatgaa aatatacaaa tgggttcttt ctggtaaaaa caaagaatat
     1081 taaattcaat gtaatacaac cgtgaatggt taatccgagt agaacagtaa ccatatacat
     1141 tcccaatgaa gaagctgtat ttgcaaggtc tttgatctcg agaatcttac caacaattaa
     1201 aaagaaaata ccaattggtg agtaccacat aattaactta actaatttca ttacaacttc
     1261 attcaagaca ccgaaaaatt cgaacattat tcttgcttcc tttcccattt gacaaggtct
     1321 ccctttagtg agggttaatt ctccctatag tgagtcgtat taggatccgg cgcgcctagc
     1381 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgacgcgtc gatttcggcc
     1441 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaattcag
     1501 ggcgcaaggg ctgctaaagg aagcggaaca cgtagaaagc cagtccgcag aaacggtgct
     1561 gaccccggat gaatgtcagc tactgggcta tctggacaag ggaaaacgca agcgcaaaga
     1621 gaaagcaggt agcttgcagt gggcttacat ggcgatagct agactgggcg gttttatgga
     1681 cagcaagcga accggaattg ccagctgggg cgccctctgg taaggttggg aagccctgca
     1741 aagtaaactg gatggctttc ttgccgccaa tgatctgatg gcgcagggga tcaagatctg
     1801 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     1861 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     1921 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     1981 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     2041 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     2101 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atcccacctt gctcctgccg
     2161 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     2221 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     2281 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     2341 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     2401 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     2461 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     2521 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     2581 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgaattgaa aaaggaagag
     2641 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     2701 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     2761 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     2821 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     2881 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     2941 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     3001 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     3061 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     3121 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     3181 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     3241 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     3301 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     3361 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     3421 cacgacgggg agccaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3481 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     3541 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     3601 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     3661 caa
//
