LOCUS       sequence-75925-e         599 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Subcloning vector for Golden Gate-based site-directed mutagenesis; 
            BglII and KpnI to release the insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 599)
  AUTHORS   Kirchmaier S, Lust K, Wittbrodt J
  TITLE     Golden GATEway cloning--a combinatorial approach to generate fusion 
            and recombination constructs.
  JOURNAL   PLoS One. 2013 Oct 7;8(10):e76117. doi: 
            10.1371/journal.pone.0076117.
  PUBMED    24116091
REFERENCE   2  (bases 1 to 599)
  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: "PLoS One.";
            date: "2013-10-7"; volume: "8(10):e76117. doi"; pages: " 
            10.1371/journal.pone.0076117"
COMMENT     Sequence Label: pGGW_SM_BglII
FEATURES             Location/Qualifiers
     source          1..599
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    29..128
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(170..553)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASNALGALPYGRPAGGREFTSDIEFPR
                     PPWRPGACDVGPNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEA
                     RTDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     319..341
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     334..350
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     357..376
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        357..375
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      406..415
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      410..419
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(513..531)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(514..531)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(549..565)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(549..565)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(562..584)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    573..589
                     /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)."
ORIGIN
        1 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
       61 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      121 agcaggctcc gcggccgccc ccttcaccag atctgtgaga gacccgcgct taatgcgccg
      181 ctacagggcg cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
      241 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
      301 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattgtaat
      361 acgactcact atagggcgaa ttgggcccga cgtcgcatgc tcccggccgc catggcggcc
      421 gcgggaattc gatatcacta gtgaattcgc ggccgcctgc aggtcgacca tatgggagag
      481 ctcccaacgc gttggatgca tagcttgagt attctatagt gtcacctaaa tagcttggcg
      541 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacac
//
