LOCUS       sequence-75912-e         818 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Golden Gate entry vector at position 4, LacZ, final entry vector for
            the assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 818)
  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 818)
  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_4'_XcmI-LacZ
FEATURES             Location/Qualifiers
     source          1..818
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        54..72
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             complement(110..493)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASNALGALPYGRPAGGREFTSDIEFPR
                     PPWRPGACDVGPNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEA
                     RTDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     259..281
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     273..290
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     274..290
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     297..316
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        297..315
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      346..355
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      350..359
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(453..471)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(489..505)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(489..505)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(502..524)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    513..529
                     /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(537..543,544..561,562..567))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    582..603
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(720..737)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcatgctcgt ttagggaacc gccattctgc ctggggacgt cggagcaagc ttgatttagg
       61 tgacactata gaggtctcaa ctaccggatc cagcggtggc gtggcgcgct taatgcgccg
      121 ctacagggcg cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
      181 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
      241 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattgtaat
      301 acgactcact atagggcgaa ttgggcccga cgtcgcatgc tcccggccgc catggcggcc
      361 gcgggaattc gatatcacta gtgaattcgc ggccgcctgc aggtcgacca tatgggagag
      421 ctcccaacgc gttggatgca tagcttgagt attctatagt gtcacctaaa tagcttggcg
      481 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
      541 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
      601 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
      661 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
      721 tcgctcactg actcgctgcg ctcgccacgc caccgctggt acctaagtga gaccctcatc
      781 ctcccgggta actaagtaag gatctcgacc gatcctga
//
