LOCUS       sequence-134323-         804 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses two gRNAs targeting the TFRC promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 804)
  AUTHORS   Aparicio-Prat E, Arnan C, Sala I, Bosch N, Guigo R, Johnson R
  TITLE     DECKO: Single-oligo, dual-CRISPR deletion of genomic elements 
            including long non-coding RNAs.
  JOURNAL   BMC Genomics. 2015 Oct 23;16(1):846. doi: 10.1186/s12864-015-2086-z.
  PUBMED    26493208
REFERENCE   2  (bases 1 to 804)
  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: "BMC 
            Genomics."; date: "2015-10-23"; volume: "16(1):846. doi"; pages: " 
            10.1186/s12864-015-2086-z"
COMMENT     Sequence Label: pDECKO_TFRC_C (with mCherry)
FEATURES             Location/Qualifiers
     source          1..804
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      91..100
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(107..160)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(170..766)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(250..270)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     747..766
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 actcgtggcc gttcacggag ccctccatgt gcaccttgaa gcgcatgaac tccttgatga
       61 tggccatgtt atcctcctcg cccttgctca ccatggtggc gaccggtggg ccaggattct
      121 cctcgacgtc accgcatgtt agcagacttc ctctgccctc tcccgatccg gcaccgggct
      181 tgcgggtcat gcaccaggtg cgcggtcctt cgggcacctc gacgtcggcg gtgacggtga
      241 agccgagccg ctcgtagaag gggaggttgc ggggcgcgga ggtctccagg aaggcgggca
      301 ccccggcgcg ctcggccgcc tccactccgg ggagcacgac ggcgctgccc agacccttgc
      361 cctggtggtc gggcgagact ccgacggtgg ccaggaacca cgcgggctcc ttgggccggt
      421 gcggcgccag gaggccttcc atctgttgct gcgcggccag ccgggaaccg ctcaactcgg
      481 ccatgcgcgg gccgatctcg gcgaacaccg cccccgcttc gacgctctcc ggcgtggtcc
      541 agaccgccac ggcggcgccg tcgtccgcga cccacacctt gccgatgtcg agcccgacgc
      601 gcgtgaggaa gagttcttgc agctcggtga cccgctcgat gtggcggtcc ggatcgacgg
      661 tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag ggtgcgtacg gccctgggga
      721 cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc ggtcatggtg gccgtacgtc
      781 acgacacctg aaatggaaga aaaa
//
