LOCUS       sequence-163314-         778 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 778)
  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 778)
  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-BC5
FEATURES             Location/Qualifiers
     source          1..778
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(81..527)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASSLVPSSDPLVTAASVLEFALVSSGF
                     SSEIDRKSGSAGMPRPISYAVFCLRANSADIHHTGGRSSMHLEGPIRPIVSRITIHWPS
                     FYNVVTGKTLALPNLIALQHIPLSPAGVIAKRPAPIALPNSCAA"
     primer_bind     195..217
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     209..226
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     210..226
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     233..252
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        233..251
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(487..505)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(488..505)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(523..539)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(523..539)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(536..558)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    547..563
                     /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(571..577,578..595,596..601))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    616..637
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(754..771)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
       61 agggcgcgtc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
      121 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
      181 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgtaatacga
      241 ctcactatag ggcgaattgg gccctctaga tgcatgctcg agcggccgcc agtgtgatgg
      301 atatctgcag aattcgccct taagcagaag acggcatacg agatcggtct cggcattcct
      361 gctgaaccgc tcttccgatc tatctcacta gagaacccac tgcttacaag ggcgaattcc
      421 agcacactgg cggccgttac tagtggatcc gagctcggta ccaagcttga tgcatagctt
      481 gagtattcta tagtgtcacc taaatagctt ggcgtaatca tggtcatagc tgtttcctgt
      541 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
      601 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
      661 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
      721 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcg
//
