LOCUS       sequence-75343-e        3020 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid compatible with the Golden Gate TALEN cloning system. To 
            clone TAL effectors 5.5 RVDs long. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3020)
  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 3020)
  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: pFUSC5
FEATURES             Location/Qualifiers
     source          1..3020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             427..1218
                     /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      1311..1899
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1800..1819
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2053..2070
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2229..2256)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2348..2434)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2488..2865)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLDSH
                     HLVRGQVQGETLKTGRRFTTS"
     primer_bind     2498..2514
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2730..2746
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2756..2775
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2756..2774
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2822..2842)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2822..2840)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2861..2877)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2861..2877)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2874..2896)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2885..2901
                     /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(2909..2915,2916..2933,2934..2939))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2954..2975
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gcggctgttg ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat
       61 cgagacgtct agaccagcca ggacagaaat gcctcgactt cgctgctacc caaggttgcc
      121 gggtgacgca caccgtggaa acggatgaag gcacgaaccc agtggacata agcctgttcg
      181 gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc
      241 gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt
      301 ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt
      361 ttgatgttat ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta
      421 aacattatga gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc
      481 gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg
      541 gatggcggcc tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt
      601 gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga
      661 gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg
      721 tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt
      781 gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca
      841 agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct
      901 gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac
      961 tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta
     1021 accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc
     1081 cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc
     1141 ttggcctcgc gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc
     1201 aaggtagtcg gcaaataacc ctcgagccac ccatgaccaa aatcccttaa cgtgagttac
     1261 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1321 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1381 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     1441 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     1501 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     1561 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     1621 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     1681 ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg
     1741 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     1801 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     1861 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     1921 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     1981 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     2041 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     2101 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     2161 aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta
     2221 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt
     2281 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc
     2341 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     2401 cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg
     2461 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaag
     2521 cgtctccccc tgaacctgac cccggaccaa gtggtggcta tcgagaccgg cgccgctaca
     2581 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     2641 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     2701 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata
     2761 cgactcacta tagggcgaat tgggtaccgg gccccccctc gaggtcctcc agcttttgtt
     2821 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     2881 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     2941 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3001 tccaccggtg gtctcgtgca
//
