LOCUS       sequence-79884-e         480 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  CAG promoter-driven expression vector suitable for ligation of 
            multiple inserts by Golden Gate Cloning using Esp3I.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 480)
  AUTHORS   Hermann M, Stillhard P, Wildner H, Seruggia D, Kapp V, 
            Sanchez-Iranzo H, Mercader N, Montoliu L, Zeilhofer HU, Pelczar P
  TITLE     Binary recombinase systems for high-resolution conditional 
            mutagenesis.
  JOURNAL   Nucleic Acids Res. 2014 Jan 9.
  PUBMED    24413561
REFERENCE   2  (bases 1 to 480)
  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: "Nucleic 
            Acids Res. 2014 Jan 9."
COMMENT     Sequence Label: pCAG-Golden-Gate-Esp3I-Destination
FEATURES             Location/Qualifiers
     source          1..480
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        40..58
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(52..357)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLRWRVDPL"
     primer_bind     207..229
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     221..238
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     222..238
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        248..266
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(314..334)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(314..332)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(353..369)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(353..369)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(366..388)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    377..393
                     /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(401..407,408..425,426..431))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    446..467
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gttcatgcct tcttcttttt cctacagatc cttaattaat aatacgactc actatagggg
       61 gtcgacccgc caccggagac gccattcgcc attcaggctg cgcaactgtt gggaagggcg
      121 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
      181 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
      241 gcgcgcgtaa tacgactcac tatagggcga attgggtacc gggccccccc tcgaggtcct
      301 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
      361 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
      421 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgcc
//
