LOCUS       sequence-163298-         777 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 777)
  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 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: "G3 
            (Bethesda)."; date: "2023-09-30"; volume: "13(10):jkad162. doi"; 
            pages: " 10.1093/g3journal/jkad162"
COMMENT     Sequence Label: TOPO-BC48
FEATURES             Location/Qualifiers
     source          1..777
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(76..522)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASSLVPSSDPLVTAASVLEFALVSSGF
                     SSCTDRKSGSAGMPRPISYAVFCLRANSADIHHTGGRSSMHLEGPIRPIVSRITIHWPS
                     FYNVVTGKTLALPNLIALQHIPLSPAGVIAKRPAPIALPNSCAA"
     primer_bind     190..212
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     204..221
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     205..221
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     228..247
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        228..246
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(482..500)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(483..500)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(518..534)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(518..534)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(531..553)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    542..558
                     /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(566..572,573..590,591..596))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    611..632
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(749..766)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
       61 gcgtccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
      121 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
      181 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa tacgactcac
      241 tatagggcga attgggccct ctagatgcat gctcgagcgg ccgccagtgt gatggatatc
      301 tgcagaattc gcccttaagc agaagacggc atacgagatc ggtctcggca ttcctgctga
      361 accgctcttc cgatctgtac aactagagaa cccactgctt acaagggcga attccagcac
      421 actggcggcc gttactagtg gatccgagct cggtaccaag cttgatgcat agcttgagta
      481 ttctatagtg tcacctaaat agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
      541 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
      601 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
      661 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
      721 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcg
//
