LOCUS       sequence-75341-e        2995 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid compatible with the Golden Gate TALEN cloning system. To 
            clone TAL effectors 3.5 RVDs long. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2995)
  AUTHORS   Boissel S, Jarjour J, Astrakhan A, Adey A, Gouble A, Duchateau P, 
            Shendure J, Stoddard BL, Certo MT, Baker D, Scharenberg AM
  TITLE     megaTALs: a rare-cleaving nuclease architecture for therapeutic 
            genome engineering.
  JOURNAL   Nucleic Acids Res. 2013 Nov 26.
  PUBMED    24285304
REFERENCE   2  (bases 1 to 2995)
  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. 2013 Nov 26."
COMMENT     Sequence Label: pFUSC3
FEATURES             Location/Qualifiers
     source          1..2995
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             402..1193
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1286..1874
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1775..1794
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2028..2045
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2204..2231)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2323..2409)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2463..2840)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLDSH
                     HLVRGQVQGETLKTGRRFTTS"
     primer_bind     2473..2489
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2705..2721
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2731..2750
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2731..2749
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2797..2817)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2797..2815)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2836..2852)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2836..2852)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2849..2871)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2860..2876
                     /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(2884..2890,2891..2908,2909..2914))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2929..2950
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gaccatggcc tgaccccgga ccaagtggtg gctatcgaga cgtctagacc agccaggaca
       61 gaaatgcctc gacttcgctg ctacccaagg ttgccgggtg acgcacaccg tggaaacgga
      121 tgaaggcacg aacccagtgg acataagcct gttcggttcg taagctgtaa tgcaagtagc
      181 gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag
      241 tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg cctcgggcat
      301 ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg
      361 ttacgcagca gggcagtcgc cctaaaacaa agttaaacat tatgagggaa gcggtgatcg
      421 ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga
      481 cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagtg
      541 atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga
      601 tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag
      661 aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac
      721 tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga
      781 tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag
      841 gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa
      901 atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag
      961 tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg
     1021 tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag
     1081 ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg
     1141 aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa taaccctcga
     1201 gccacccatg accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc
     1261 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     1321 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     1381 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     1441 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     1501 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     1561 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     1621 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagcatt
     1681 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     1741 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     1801 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     1861 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     1921 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     1981 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     2041 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     2101 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     2161 ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag
     2221 gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc
     2281 tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag
     2341 agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg
     2401 ttttatttga tgcctggcag ttccctactc tcgcgttaac gctagcatgg atgttttccc
     2461 agtcacgacg ttgtaaaacg acggccagtc ttaagcgtct ccccctgaac ctgaccccgg
     2521 accaagtggt ggctatcgag accggcgccg ctacagggcg cgtcccattc gccattcagg
     2581 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     2641 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     2701 cgttgtaaaa cgacggccag tgagcgcgcg taatacgact cactataggg cgaattgggt
     2761 accgggcccc ccctcgaggt cctccagctt ttgttccctt tagtgagggt taattgcgcg
     2821 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     2881 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     2941 actcacatta attgcgttgc gctcactgcc cgctttccac cggtggtctc gccag
//
