LOCUS       sequence-72656-e        1127 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of TALEN proteins with FokI-DDD domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1127)
  AUTHORS   Hu R, Wallace J, Dahlem TJ, Grunwald DJ, O'Connell RM
  TITLE     Targeting human microRNA genes using engineered Tal-effector 
            nucleases (TALENs).
  JOURNAL   PLoS One. 2013 May 7;8(5):e63074. doi: 10.1371/journal.pone.0063074.
            Print 2013.
  PUBMED    23667577
REFERENCE   2  (bases 1 to 1127)
  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: "2013-05-7"; volume: "8(5):e63074. doi"; pages: " 
            10.1371/journal.pone.0063074. Print 2013"
COMMENT     Sequence Label: pCS2TAL3-DDD
FEATURES             Location/Qualifiers
     source          1..1127
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        63..81
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     63..80
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      113..122
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             125..145
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             146..211
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(518..925)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLQGG
                     TRQCIARCMHCLHCGHAATFCNLHKLACVQL"
     primer_bind     775..797
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     789..806
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     790..806
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     816..835
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        816..834
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(882..902)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(882..900)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(921..937)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(921..937)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(934..956)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    945..961
                     /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(969..975,976..993,994..999))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1014..1035
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ctatataagc aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct
       61 tgatttaggt gacactatag aatacaagct acttgttctt tttgcaggat ctgccaccat
      121 ggctccaaag aagaagcgta aggtagacta caaagaccat gacggtgatt ataaagatca
      181 tgacatcgat tacaaggatg acgatgacaa gggtaccgtg gatctacgca cgctcggcta
      241 cagtcagcag cagcaagaga agatcaaacc gaaggtgcgt tcgacagtgg cgcagcacca
      301 cgaggcactg gtgggccatg ggtttacaca cgcgcacatc gttgcgctca gccaacaccc
      361 ggcagcgtta gggaccgtcg ctgtcacgta tcagcacata atcacggcgt tgccagaggc
      421 gacacacgaa gacatcgttg gcgtcggcaa acagtggtcc ggcgcacgcg ccctggaggc
      481 cttgctcacg gatgcggggg agttgagagg tccgccgtta cagttggaca caggccaact
      541 tgtgaagatt gcaaaacgtg gcggcgtgac cgcaatggag gcagtgcatg catcgcgcaa
      601 tgcactgacg ggtgcccccc tggagacggg cgccgctaca gggcgcgtcc cattcgccat
      661 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
      721 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
      781 cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat
      841 tgggtaccgg gccccccctc gaggtcctcc agcttttgtt ccctttagtg agggttaatt
      901 gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
      961 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     1021 agctaactca cattaattgc gttgcgctca ctgcccgctt tccaccggtc gtctccaacg
     1081 accacctcgt cgccttggcc tgcctcggcg gacgtcctgc catggat
//
