LOCUS       sequence-170886-        1575 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Express  LdPBK_241510.1 and LdMT targeting  gRNAs simutaneously.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1575)
  AUTHORS   Zhang WW, Lypaczewski P, Matlashewski G
  TITLE     Optimized CRISPR-Cas9 Genome Editing for Leishmania and Its Use To 
            Target a Multigene Family, Induce Chromosomal Translocation, and 
            Study DNA Break Repair Mechanisms.
  JOURNAL   mSphere. 2017 Jan 18;2(1). pii: e00340-16. doi: 
            10.1128/mSphere.00340-16. eCollection 2017 Jan-Feb.
  PUBMED    28124028
REFERENCE   2  (bases 1 to 1575)
  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: "mSphere."; 
            date: "2017-01-18"; volume: "2(1). pii: e00340-16. doi"; pages: " 
            10.1128/mSphere.00340-16. eCollection 2017 Jan-Feb"
COMMENT     Sequence Label: pSPneogRNA241510+MT
FEATURES             Location/Qualifiers
     source          1..1575
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        1..19
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1..18
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_RNA        236..311
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           318..386
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     ncRNA           399..435
                     /product="self-cleaving hammerhead ribozyme (Ferre-D'Amare 
                     and Scott, 2010)"
                     /label=hammerhead ribozyme
                     /ncRNA_class="hammerhead_ribozyme"
     misc_RNA        461..536
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           543..610
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     CDS             732..1526
                     /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
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHKAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(786..805)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1396..1415
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1556..1575)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1557..1575)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 atttaggtga cactatagaa ctcgagcagc tgaagcttgt gagttatgag gtctgcgatt
       61 gacgtaggag ttgcaagggg gagggggtat gaacggggtg ggtagagctt ttttaggtgg
      121 aagtagtgag agggtgggct tgtgagaatt tgaggtgtgt tcgtgatgtg tggatcttat
      181 cgggggctcg gttgagtttt tggtttggtg attttgtggt gtcgttctgc agccagtttt
      241 agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac
      301 cgagtcggtg cttttttggc cggcatggtc ccagcctcct cgctggcgcc ggctgggcaa
      361 catgcttcgg ccatggcgaa tgggacggat ctcaccatct gatgagtccg tgaggacgaa
      421 acgagtaagc tcgtcatggt ggtccaggcg cagtggatgc gttttagagc tagaaatagc
      481 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
      541 ttggccggca tggtcccagc ctcctcgctg gcgccggctg ggcaacatgc ttcggcatgg
      601 cgaatgggac ggatccccgg gtaccgagct ctctcttctc cttcccctct ctctcctctc
      661 tcctctctcc agagctctct cttctccttc ccctctctct cctctctcct ctctccagag
      721 ctcgaattcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
      781 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
      841 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
      901 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
      961 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     1021 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     1081 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccacaaag
     1141 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     1201 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     1261 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     1321 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     1381 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     1441 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     1501 atcgccttct tgacgagttc ttctgatcga tgatatcaga tcgatctgcc ggtctcccta
     1561 tagtgagtcg tatta
//
