LOCUS       sequence-163635-         814 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 814)
  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 814)
  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-BC41
FEATURES             Location/Qualifiers
     source          1..814
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(80..526)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASSLVPSSDPLVTAASVLEFALVSSGF
                     SSHEDRKSGSAGMPRPISYAVFCLRANSADIHHTGGRSSMHLEGPIRPIVSRITIHWPS
                     FYNVVTGKTLALPNLIALQHIPLSPAGVIAKRPAPIALPNSCAA"
     primer_bind     194..216
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     208..225
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     209..225
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     232..251
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        232..250
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(486..504)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(487..504)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(522..538)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(522..538)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(535..557)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    546..562
                     /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(570..576,577..594,595..600))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    615..636
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(753..770)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
       61 gggcgcgtcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
      121 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
      181 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac
      241 tcactatagg gcgaattggg ccctctagat gcatgctcga gcggccgcca gtgtgatgga
      301 tatctgcaga attcgccctt aagcagaaga cggcatacga gatcggtctc ggcattcctg
      361 ctgaaccgct cttccgatct tcatggctag agaacccact gcttacaagg gcgaattcca
      421 gcacactggc ggccgttact agtggatccg agctcggtac caagcttgat gcatagcttg
      481 agtattctat agtgtcacct aaatagcttg gcgtaatcat ggtcatagct gtttcctgtg
      541 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
      601 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
      661 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
      721 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
      781 gttcggctgc ggcgagcggt atcagctcac tcaa
//
