LOCUS       sequence-63268-e         652 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Destination vector for Voytas Golden Gate TALEN assembly 
            incorporating a CAG promoter for mammalian expression and a T7 
            promoter for synthesis of TALEN mRNA. Uses homodimeric FokI 
            domains..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 652)
  AUTHORS   Hermann M, Cermak T, Voytas DF, Pelczar P
  TITLE     Mouse genome engineering using designer nucleases.
  JOURNAL   J Vis Exp. 2014 Apr 2;(86). doi: 10.3791/50930.
  PUBMED    24747757
REFERENCE   2  (bases 1 to 652)
  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: "J Vis 
            Exp."; date: "2014-04-2"; volume: "(86). doi"; pages: " 
            10.3791/50930"
COMMENT     Sequence Label: pCAG-T7-TALEN(Sangamo)-Destination
FEATURES             Location/Qualifiers
     source          1..652
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(30..437)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLQGG
                     TRQCIARCMHCLHCGHAATFCNLHKLACVQL"
     primer_bind     287..309
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     301..318
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     302..318
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     328..347
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        328..346
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(394..414)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(394..412)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(433..449)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(433..449)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(446..468)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    457..473
                     /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(481..487,488..505,506..511))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    526..547
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cggatgcggg ggagttgaga ggtccgccgt tacagttgga cacaggccaa cttgtgaaga
       61 ttgcaaaacg tggcggcgtg accgcaatgg aggcagtgca tgcatcgcgc aatgcactga
      121 cgggtgcccc cctggagacg ggcgccgcta cagggcgcgt cccattcgcc attcaggctg
      181 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
      241 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
      301 tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga attgggtacc
      361 gggccccccc tcgaggtcct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt
      421 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
      481 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
      541 cacattaatt gcgttgcgct cactgcccgc tttccaccgg tcgtctccaa cgaccacctc
      601 gtcgccttgg cctgcctcgg cggacgtcct gccatggatg cagtgaaaaa gg
//
