LOCUS       sequence-75347-e        3031 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid compatible with the Golden Gate TALEN cloning system. To 
            clone TAL effectors 9.5 RVDs long..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3031)
  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 3031)
  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: pFUSC9
FEATURES             Location/Qualifiers
     source          1..3031
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             438..1229
                     /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      1322..1910
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1811..1830
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2064..2081
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2240..2267)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2359..2445)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2499..2876)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLDSH
                     HLVRGQVQGETLKTGRRFTTS"
     primer_bind     2509..2525
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2741..2757
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2767..2786
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2767..2785
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2833..2853)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2833..2851)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2872..2888)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2872..2888)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2885..2907)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2896..2912
                     /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(2920..2926,2927..2944,2945..2950))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2965..2986
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac cccggaccaa
       61 gtggtggcta tcgagacgtc tagaccagcc aggacagaaa tgcctcgact tcgctgctac
      121 ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc cagtggacat
      181 aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc aactggtcca
      241 gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg
      301 actgtttttt tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg
      361 tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta
      421 aaacaaagtt aaacattatg agggaagcgg tgatcgccga agtatcgact caactatcag
      481 aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg
      541 gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg gttacggtga
      601 ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg
      661 cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg
      721 acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca
      781 atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc
      841 tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg
      901 atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact
      961 cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt
     1021 acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc
     1081 gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag
     1141 aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac tacgtgaaag
     1201 gcgagatcac caaggtagtc ggcaaataac cctcgagcca cccatgacca aaatccctta
     1261 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     1321 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     1381 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     1441 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     1501 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     1561 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     1621 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     1681 cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag
     1741 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     1801 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     1861 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     1921 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     1981 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     2041 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     2101 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     2161 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc gctagccagg
     2221 aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg
     2281 cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt
     2341 caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat
     2401 aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc
     2461 ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt aaaacgacgg
     2521 ccagtcttaa gcgtctcccc ctgaacctga ccccggacca agtggtggct atcgagaccg
     2581 gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     2641 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     2701 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag
     2761 cgcgcgtaat acgactcact atagggcgaa ttgggtaccg ggccccccct cgaggtcctc
     2821 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     2881 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     2941 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3001 actgcccgct ttccaccggt ggtctccgct c
//
