LOCUS       sequence-123701-         772 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  It encodes a fluorescent protein that can be fused to a gene of 
            interest using SEC CRISPR strategy.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 772)
  TITLE     Hobert plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 772)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pUA97
FEATURES             Location/Qualifiers
     source          1..772
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             242..547
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPG*RMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     426..445
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(595..618)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(622..645)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(649..672)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    691..724
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 tgtcaaaaag aggtatgcta tgaagcagcg tattacagtg acagttgaca gcgacagcta
       61 tcagttgctc aaggcatata tgatgtcaat atctccggtt cggtaagcac aaccatgcag
      121 aatgaagccc gtcgtctgcg tgccgaacgc tggaaagcgg aaaatcagga agggatggct
      181 gaggtcgccc ggtttattga aatgaacggc tcttttgccg acgagaacag gggctggtga
      241 aatgcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt
      301 acagagtgat attattgaca cgcccgggtg acggatggtg atccccctgg ccagtgcacg
      361 tctgctgtca gataaagtcc cccgtgaact ttacccggtg gtgcatatcg gggatgaaag
      421 ctggcgcatg atgaccaccg atatggccag tgtgccggtc tccgttatcg gggaagaagt
      481 ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg
      541 aatataaatg tcaggctccc ttatacacag ccagtctgca ggtcgaacta gtctcttgtc
      601 atcgtcatcc ttgtaatcac gcttgtcgtc gtcgtccttg tagtcacgct tatcgtcatc
      661 gtctttataa tcagatccgg ctccctgaaa ataacttcgt ataatgtatg ctatacgaag
      721 ttattaaaaa ttgcggtcat aaactgaaac gtaacatatg ataaggtatt tt
//
