LOCUS       sequence-76030-e         686 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Destination vector for Golden Gate assembly; 5' entry vector for 
            Gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 686)
  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 686)
  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: p5E_GGWDest+
FEATURES             Location/Qualifiers
     source          1..686
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     8..24
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    64..159
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      177..186
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(199..582)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASNALGALPYGRPAGGREFTSDIEFPR
                     PPWRPGACDVGPNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEA
                     RTDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     363..379
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     386..405
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        386..404
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      435..444
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      439..448
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(542..560)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(543..560)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(578..594)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(578..594)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(591..613)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    602..618
                     /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(626..632,633..650,651..656))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 cgacgttgta aaacgacggc cagtcttaag ctcgggcccg agttaacgct accatggagc
       61 tccaaataat gattttattt tgactgatag tgacctgttc gttgcaacaa attgataagc
      121 aatgcttttt tataatgcca actttgtata gaaaagttga gggcgaattg ggtaccgccg
      181 ccatggcgag acccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct
      241 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      301 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      361 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tgggcccgac
      421 gtcgcatgct cccggccgcc atggcggccg cgggaattcg atatcactag tgaattcgcg
      481 gccgcctgca ggtcgaccat atgggagagc tcccaacgcg ttggatgcat agcttgagta
      541 ttctatagtg tcacctaaat agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
      601 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
      661 ggggtgccta atgagtgagc taactc
//
