LOCUS       sequence-77102-e         728 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Destination vector for Golden Gate assembly; Few restriction sites 
            for subcloning..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 728)
  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 728)
  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: pGGDestSC+ATG
FEATURES             Location/Qualifiers
     source          1..728
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     5..22
                     /label=L4440
                     /note="L4440 vector, forward primer"
     regulatory      117..126
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(139..522)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASNALGALPYGRPAGGREFTSDIEFPR
                     PPWRPGACDVGPNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEA
                     RTDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     288..310
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     302..319
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     303..319
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     326..345
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        326..344
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      375..384
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      379..388
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(482..500)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(483..500)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(518..534)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(518..534)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(531..553)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    542..558
                     /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(566..572,573..590,591..596))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    611..632
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
       61 cgcgcgttgg ccgattcatt aatgcagtat aggcgcgcca gggcgaattg ggtaccgccg
      121 ccatggcgag acccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct
      181 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      241 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      301 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tgggcccgac
      361 gtcgcatgct cccggccgcc atggcggccg cgggaattcg atatcactag tgaattcgcg
      421 gccgcctgca ggtcgaccat atgggagagc tcccaacgcg ttggatgcat agcttgagta
      481 ttctatagtg tcacctaaat agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
      541 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
      601 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
      661 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
      721 gtttgcgt
//
