LOCUS       sequence-80323-e         817 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Contain one loxP-PGK-Neo-pA-loxP cassette, used for golden-gate 
            reaction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 817)
  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 817)
  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-Neo
FEATURES             Location/Qualifiers
     source          1..817
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    105..126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(141..146,147..164,165..171)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    179..195
                     /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     184..206
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     203..219
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     203..219
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        237..255
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     237..254
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    complement(380..413)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(460..483)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
       61 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      121 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      181 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctattt
      241 aggtgacact atagaatact caagctatgc atcaagcttg gtaccgagct cggatccact
      301 agtaacggcc gccagtgtgc tggaattcgc ccttaggtct cacgtgctaa agggaacaaa
      361 agctggagct ccaccgcgga taacttcgta tagcatacat tatacgaagt tatcgcgccc
      421 taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg
      481 ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt
      541 aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta
      601 gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta
      661 gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc
      721 ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga
      781 aggggtgggt ccgggggcgg gctcaggggc gggctca
//
