LOCUS       sequence-76025-e         841 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 841)
  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 841)
  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..841
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     11..27
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    67..162
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(196..579)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASNALGALPYGRPAGGREFTSDIEFPR
                     PPWRPGACDVGPNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEA
                     RTDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     360..376
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     383..402
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        383..401
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      432..441
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      436..445
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(539..557)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(540..557)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(575..591)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(575..591)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(588..610)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    599..615
                     /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(623..629,630..647,648..653))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    668..689
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(806..823)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc ccgagttaac gctaccatgg
       61 agctccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgata
      121 agcaatgctt ttttataatg ccaactttgt atagaaaagt tgagggcgaa ttgggtaccg
      181 tatcgagacc cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg
      241 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      301 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      361 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gcccgacgtc
      421 gcatgctccc ggccgccatg gcggccgcgg gaattcgata tcactagtga attcgcggcc
      481 gcctgcaggt cgaccatatg ggagagctcc caacgcgttg gatgcatagc ttgagtattc
      541 tatagtgtca cctaaatagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
      601 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
      661 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
      721 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
      781 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gggtctctta
      841 a
//
