LOCUS       sequence-75920-e         636 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Golden Gate entry vector at position 8, LacZ, final entry vector for
            the assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 636)
  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 636)
  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_8'_XcmI-LacZ
FEATURES             Location/Qualifiers
     source          1..636
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        28..46
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             complement(84..467)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASNALGALPYGRPAGGREFTSDIEFPR
                     PPWRPGACDVGPNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEA
                     RTDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     233..255
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     247..264
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     248..264
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     271..290
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        271..289
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      320..329
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      324..333
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(427..445)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(463..479)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(463..479)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(476..498)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    487..503
                     /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(511..517,518..535,536..541))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    556..577
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ctgcctgggg acgtcggagc aagcttgatt taggtgacac tatagaggtc tcagccggcg
       61 gatccagcgg tggcgtggcg cgcttaatgc gccgctacag ggcgcgtcca ttcgccattc
      121 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      181 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      241 cgacgttgta aaacgacggc cagtgaattg taatacgact cactataggg cgaattgggc
      301 ccgacgtcgc atgctcccgg ccgccatggc ggccgcggga attcgatatc actagtgaat
      361 tcgcggccgc ctgcaggtcg accatatggg agagctccca acgcgttgga tgcatagctt
      421 gagtattcta tagtgtcacc taaatagctt ggcgtaatca tggtcatagc tgtttcctgt
      481 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
      541 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
      601 tttccagtcg ggaaacctgt cgtgccagct gcatta
//
