LOCUS       sequence-80320-e         881 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Contain one loxP-PGK-Hyg-pA-loxP cassette, used for golden-gate 
            reaction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 881)
  AUTHORS   Luo Y, Lin L, Bolund L, Sorensen CB
  TITLE     Efficient construction of rAAV-based gene targeting vectors by 
            Golden Gate cloning.
  JOURNAL   Biotechniques. 2014 May 1;56(5):263-8. doi: 10.2144/000114169. 
            eCollection 2014 May.
  PUBMED    24806227
REFERENCE   2  (bases 1 to 881)
  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"; doi: "10.2144/000114169";
            journalName: "Biotechniques"; date: "2014-05-1- 1"; volume: "56"; 
            issue: "5"; pages: "263-8"
COMMENT     Sequence Label: pGolden-Hyg
FEATURES             Location/Qualifiers
     source          1..881
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    109..130
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(145..150,151..168,169..175)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    183..199
                     /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)."
     primer_bind     188..210
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     207..223
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     207..223
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        241..259
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     241..258
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    351..384
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    409..633
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     622..639
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
ORIGIN
        1 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
       61 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
      121 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
      181 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
      241 atttaggtga cactatagaa tactcaagct atgcatcaag cttggtaccg agctcggatc
      301 cactagtaac ggccgccagt gtgctggaat tcgcccttag gtctctggac ataacttcgt
      361 ataatgtatg ctatacgaag ttatagctgg ttctttccgc ctcagaagcc atagagccca
      421 ccgcatcccc agcatgcctg ctattgtctt cccaatcctc ccccttgctg tcctgcccca
      481 ccccaccccc cagaatagaa tgacacctac tcagacaatg cgatgcaatt tcctcatttt
      541 attaggaaag gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc
      601 aaacaacaga tggctggcaa ctagaaggca cagtcgaggc tgatcagcga gctctagaga
      661 tctgcaccca atcggcaggc acgggcggcg atctccaatc tgcgggatca gtcagatcac
      721 ccgagtgcgt gggcatgaca atcgtgccct ggggaccaac acaatccaga agggcctgaa
      781 tcactgcgac cggccctccc gcgacccagc cgagcgagct tagcgaactg tggacgagaa
      841 ctgtgccacc aagcgtaagg ccgttctctc gcatttgcct t
//
