LOCUS       sequence-163348-         833 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 833)
  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 833)
  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-BC20
FEATURES             Location/Qualifiers
     source          1..833
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(79..525)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASSLVPSSDPLVTAASVLEFALVSSGF
                     SRLKDRKSGSAGMPRPISYAVFCLRANSADIHHTGGRSSMHLEGPIRPIVSRITIHWPS
                     FYNVVTGKTLALPNLIALQHIPLSPAGVIAKRPAPIALPNSCAA"
     primer_bind     193..215
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     207..224
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     208..224
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     231..250
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        231..249
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(485..503)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(486..503)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(521..537)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(521..537)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(534..556)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    545..561
                     /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(569..575,576..593,594..599))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    614..635
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(752..769)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
       61 ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
      121 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
      181 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg taatacgact
      241 cactataggg cgaattgggc cctctagatg catgctcgag cggccgccag tgtgatggat
      301 atctgcagaa ttcgccctta agcagaagac ggcatacgag atcggtctcg gcattcctgc
      361 tgaaccgctc ttccgatctt taagcctaga gaacccactg cttacaaggg cgaattccag
      421 cacactggcg gccgttacta gtggatccga gctcggtacc aagcttgatg catagcttga
      481 gtattctata gtgtcaccta aatagcttgg cgtaatcatg gtcatagctg tttcctgtgt
      541 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
      601 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
      661 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
      721 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
      781 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tcc
//
