LOCUS       sequence-76031-e         747 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Destination vector for Golden Gate assembly; middle entry vector for
            Gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 747)
  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 747)
  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: pME_GGWDest+
FEATURES             Location/Qualifiers
     source          1..747
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        27..45
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     protein_bind    69..168
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     170..186
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        196..214
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      229..238
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(251..634)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASNALGALPYGRPAGGREFTSDIEFPR
                     PPWRPGACDVGPNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEA
                     RTDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     400..422
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     415..431
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        438..456
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      487..496
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      491..500
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(594..612)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(630..646)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(630..646)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(643..665)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    654..670
                     /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(678..684,685..702,703..708))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    723..744
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tgcctgggga cgtcggagca agcttgattt aggtgacact atagacggcc agtcttaagc
       61 tcgggcccca aataatgatt ttattttgac tgatagtgac ctgttcgttg caacaaattg
      121 atgagcaatg cttttttata atgccaactt tgtacaaaaa agcaggcttg taaaacgacg
      181 gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgc cgccatggcg
      241 agacccgcgc ttaatgcgcc gctacagggc gcgtccattc gccattcagg ctgcgcaact
      301 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
      361 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
      421 cgacggccag tgaattgtaa tacgactcac tatagggcga attgggcccg acgtcgcatg
      481 ctcccggccg ccatggcggc cgcgggaatt cgatatcact agtgaattcg cggccgcctg
      541 caggtcgacc atatgggaga gctcccaacg cgttggatgc atagcttgag tattctatag
      601 tgtcacctaa atagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
      661 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
      721 taatgagtga gctaactcac attaatt
//
