LOCUS       sequence-123987-         908 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses the fusion gene CAS9-HA-2xNLS-GFP in the pTREX-n backbone.
            This vector is used for cloning a specific sgRNA by BamHI, to be 
            co-expressed with Cas9 for genome editing in Trypanosoma cruzi..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 908)
  AUTHORS   Lander N, Li ZH, Niyogi S, Docampo R
  TITLE     CRISPR/Cas9-Induced Disruption of Paraflagellar Rod Protein 1 and 2 
            Genes in Trypanosoma cruzi Reveals Their Role in Flagellar 
            Attachment.
  JOURNAL   MBio. 2015 Jul 21;6(4). pii: e01012-15. doi: 10.1128/mBio.01012-15.
  PUBMED    26199333
REFERENCE   2  (bases 1 to 908)
  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: "MBio."; 
            date: "2015-07-21"; volume: "6(4). pii: e01012-15. doi"; pages: " 
            10.1128/mBio.01012-15"
COMMENT     Sequence Label: Cas9/pTREX-n
FEATURES             Location/Qualifiers
     source          1..908
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(join(3..710,711..713))
                     /codon_start=1
                     /product="enhanced GFP (EGFP) derivative that exhibits 
                     robust folding and enhanced photostability (Zhong et al., 
                     2019)"
                     /label=SiriusGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELY"
     primer_bind     complement(42..63)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     650..671
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(753..773)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(780..800)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(813..839)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             complement(819..845)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
ORIGIN
        1 ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc
       61 atgtgatcgc gcttctcgtt ggggtctttg ctcagcttgg actgggtgct caggtagtgg
      121 ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg
      181 agctgcacgc tgccgtcctc gacgttgtgg cggatcttga agttcgcctt gatgccgttc
      241 ttctgcttgt cggccgtgat atagacgttg tggctgttga agttgtactc cagcttgtgc
      301 cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg
      361 gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagg tgccgtcgtc cttgaaggag
      421 atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc
      481 atgtggtcgg ggtagcggct gaagcactgc acgccgtagg tcagggtggt cacgagggtg
      541 ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgttg
      601 gtggcatcgc cctcgccctc gccgcgcacg ctgaacttgt ggccgtttac gtcgccgtcc
      661 agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct cacagatccg
      721 ctgctgccgt tgctcccact accaatgccg tccactttcc gctttttctt aggatcttcc
      781 accttgcgtt ttttcttggg ggagccgctg cccaggctgg cataatcggg cacgtcatag
      841 ggataggcgt cgcctcccag ctgagacagg tcgatccgtg tctcgtacag gccggtgatg
      901 ctctggtg
//
