LOCUS       sequence-163377-         816 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Encodes illumina barcodes for allele frequency sequencing 
            (FREQ-Seq-Squared).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 816)
  AUTHORS   Zhao R, Lukacsovich T, Gaut R, Emerson JJ
  TITLE     FREQ-Seq2: a method for precise high-throughput combinatorial 
            quantification of allele frequencies.
  JOURNAL   G3 (Bethesda). 2023 Sep 30;13(10):jkad162. doi: 
            10.1093/g3journal/jkad162.
  PUBMED    37494033
REFERENCE   2  (bases 1 to 816)
  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: "G3 
            (Bethesda)."; date: "2023-09-30"; volume: "13(10):jkad162. doi"; 
            pages: " 10.1093/g3journal/jkad162"
COMMENT     Sequence Label: TOPO-BC33
FEATURES             Location/Qualifiers
     source          1..816
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(77..523)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASSLVPSSDPLVTAASVLEFALKQKTA
                     YEIGLGIPAEPLFRSRILENPLLTRANSADIHHTGGRSSMHLEGPIRPIVSRITIHWPS
                     FYNVVTGKTLALPNLIALQHIPLSPAGVIAKRPAPIALPNSCAA"
     primer_bind     191..213
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     205..222
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     206..222
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     229..248
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        229..247
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(483..501)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(484..501)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(519..535)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(519..535)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(532..554)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    543..559
                     /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(567..573,574..591,592..597))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    612..633
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(750..767)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
       61 cgcgtccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
      121 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
      181 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattgta atacgactca
      241 ctatagggcg aattgggccc tctagatgca tgctcgagcg gccgccagtg tgatggatat
      301 ctgcagaatt cgcccttgta agcagtgggt tctctagtat ccgagatcgg aagagcggtt
      361 cagcaggaat gccgagaccg atctcgtatg ccgtcttctg cttaagggcg aattccagca
      421 cactggcggc cgttactagt ggatccgagc tcggtaccaa gcttgatgca tagcttgagt
      481 attctatagt gtcacctaaa tagcttggcg taatcatggt catagctgtt tcctgtgtga
      541 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
      601 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
      661 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
      721 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
      781 cggctgcggc gagcggtatc agctcactca aaggcg
//
