LOCUS       sequence-163367-         835 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 835)
  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 835)
  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-BC29
FEATURES             Location/Qualifiers
     source          1..835
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(75..521)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASSLVPSSDPLVTAASVLEFALVSSGF
                     SSPTDRKSGSAGMPRPISYAVFCLRANSADIHHTGGRSSMHLEGPIRPIVSRITIHWPS
                     FYNVVTGKTLALPNLIALQHIPLSPAGVIAKRPAPIALPNSCAA"
     primer_bind     189..211
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     203..220
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     204..220
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     227..246
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        227..245
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(481..499)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(482..499)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(517..533)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(517..533)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(530..552)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    541..557
                     /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(565..571,572..589,590..595))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    610..631
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(748..765)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
       61 cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
      121 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
      181 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattgtaat acgactcact
      241 atagggcgaa ttgggccctc tagatgcatg ctcgagcggc cgccagtgtg atggatatct
      301 gcagaattcg cccttaagca gaagacggca tacgagatcg gtctcggcat tcctgctgaa
      361 ccgctcttcc gatctgttgg gctagagaac ccactgctta caagggcgaa ttccagcaca
      421 ctggcggccg ttactagtgg atccgagctc ggtaccaagc ttgatgcata gcttgagtat
      481 tctatagtgt cacctaaata gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
      541 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
      601 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
      661 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
      721 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
      781 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaa
//
