LOCUS       sequence-75911-e         667 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Golden Gate entry vector at position 3, LacZ, final entry vector for
            the assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 667)
  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 667)
  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: pGGEV_3'_XcmI-LacZ
FEATURES             Location/Qualifiers
     source          1..667
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        52..70
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             complement(108..491)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASNALGALPYGRPAGGREFTSDIEFPR
                     PPWRPGACDVGPNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEA
                     RTDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     257..279
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     271..288
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     272..288
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     295..314
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        295..313
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      344..353
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      348..357
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(451..469)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(487..503)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(487..503)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(500..522)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    511..527
                     /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(535..541,542..559,560..565))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    580..601
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 atgctcgttt agggaaccgc cattctgcct ggggacgtcg gagcaagctt gatttaggtg
       61 acactataga ggtctcaacc tctggatcca gcggtggcgt ggcgcgctta atgcgccgct
      121 acagggcgcg tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
      181 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
      241 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attgtaatac
      301 gactcactat agggcgaatt gggcccgacg tcgcatgctc ccggccgcca tggcggccgc
      361 gggaattcga tatcactagt gaattcgcgg ccgcctgcag gtcgaccata tgggagagct
      421 cccaacgcgt tggatgcata gcttgagtat tctatagtgt cacctaaata gcttggcgta
      481 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
      541 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
      601 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
      661 atgaatc
//
