LOCUS       sequence-77030-e         846 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Destination vector for Golden Gate assembly; middle entry vector for
            Gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 846)
  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 846)
  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: pME_GGWDest-
FEATURES             Location/Qualifiers
     source          1..846
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    46..145
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     172..192
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        174..192
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     229..245
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     229..245
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(279..295)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(280..296)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     318..335
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    452..473
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(488..493,494..511,512..518)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    526..542
                     /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     531..553
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     550..566
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     550..566
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        584..602
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     584..601
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      696..705
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      700..709
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(739..758)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(740..758)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(765..782)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(765..781)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(774..796)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ccttacttag ttacccggga ggatgagggg atatcagctg gatggcaaat aatgatttta
       61 ttttgactga tagtgacctg ttcgttgcaa caaattgata agcaatgctt tcttataatg
      121 ccaactttgt acaagaaagc tgggtgctat gaccatgatt acgccaagcg cgcaattaac
      181 cctcactaaa gggaacaaaa gctggagctc caccgcggtg gcggccgctc tagaactagt
      241 ggatcccccg ggctgcagga attcgatatc aagcttatcg ataccgtcga cctcgagcct
      301 taagagaccc cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
      361 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
      421 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
      481 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
      541 aatttcacac aggaaacagc tatgaccatg attacgccaa gctatttagg tgacactata
      601 gaatactcaa gctatgcatc caacgcgttg ggagctctcc catatggtcg acctgcaggc
      661 ggccgcgaat tcactagtga tatcgaattc ccgcggccgc catggcggcc gggagcatgc
      721 gacgtcgggc ccaattcgcc ctatagtgag tcgtattaca attcactggc cgtcgtttta
      781 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
      841 cctttc
//
