LOCUS       sequence-22998-e         777 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 777)
  AUTHORS   Kohli M, Rago C, Lengauer C, Kinzler KW, Vogelstein B
  TITLE     Facile methods for generating human somatic cell gene knockouts 
            using recombinant adeno-associated viruses.
  JOURNAL   Nucleic Acids Res. 2004  . 32(1):e3.
  PUBMED    14704360
REFERENCE   2  (bases 1 to 777)
  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: "Nucleic 
            Acids Res. 2004  . 32(1):e3."
COMMENT     Sequence Label: pNeDaKO-Hyg
FEATURES             Location/Qualifiers
     source          1..777
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    26..59
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(89..107)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(128..144)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(128..144)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(141..163)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    152..168
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(176..182,183..200,201..206))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    221..242
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(359..376)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(610..629)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgcctccct acccggtagg gcgcgataac ttcgtataat gtatgctata cgaagttatc
       61 caccgcggtg gagctccagc ttttgttccc tttagtgagg gttaatttcg agcttggcgt
      121 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
      181 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
      241 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
      301 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
      361 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
      421 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
      481 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
      541 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
      601 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
      661 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
      721 ctcatagctc acgctgtngt atctcagttc ggtgtaggtc gcttcgctcc aagctgg
//
