LOCUS       sequence-75211-e         739 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Subcloning vector for Golden Gate-based site-directed mutagenesis; 
            BglII and KpnI to release the insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 739)
  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 739)
  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: pGGW_SM_BglII
FEATURES             Location/Qualifiers
     source          1..739
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     73..89
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        94..112
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    130..229
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     270..287
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    404..425
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(440..445,446..463,464..470)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    478..494
                     /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)."
     primer_bind     483..505
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     502..518
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        536..554
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     536..553
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      648..657
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      652..661
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(692..710)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(717..734)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(717..733)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacacg
       61 ggccagagct gccaggaaac agctatgacc atgtaatacg actcactata ggggatatca
      121 gctggatggc aaataatgat tttattttga ctgatagtga cctgttcgtt gcaacaaatt
      181 gataagcaat gctttcttat aatgccaact ttgtacaaga aagctgggtc ggcgcgccca
      241 cccttggtac cgactgagac cgcgagcgca gcgagtcagt gagcgaggaa gcggaagagc
      301 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
      361 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
      421 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
      481 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagctattta
      541 ggtgacacta tagaatactc aagctatgca tccaacgcgt tgggagctct cccatatggt
      601 cgacctgcag gcggccgcga attcactagt gatatcgaat tcccgcggcc gccatggcgg
      661 ccgggagcat gcgacgtcgg gcccaattcg ccctatagtg agtcgtatta caattcactg
      721 gccgtcgttt tacaacgtc
//
