LOCUS       sequence-105221-        3015 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Encodes RVD positions 7-9 in TALEN assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3015)
  AUTHORS   Ma AC, McNulty MS, Tanya PL, Jarryd CM, Martinez-Galvez G, Argue DP,
            Lee HB, Urban MD, Bullard CE, Blackburn PR, Man TK, Clark KJ, Ekker 
            SC
  TITLE     FusX: A rapid one-step TALE assembly system for genome science.
  JOURNAL   Hum Gene Ther. 2016 Jun;27(6):451-63. doi: 10.1089/hum.2015.172
  PUBMED    26854857
REFERENCE   2  (bases 1 to 3015)
  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"; doi: "10.1089/hum.2015"; 
            journalName: "Hum Gene Ther"; date: "2016-06"; volume: "27"; issue: 
            "6"; pages: "451-63"
COMMENT     Sequence Label: pFusX3-5
FEATURES             Location/Qualifiers
     source          1..3015
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     204..220
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(295..315)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(295..313)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(334..350)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(334..350)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(347..369)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    358..374
                     /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(382..388,389..406,407..412))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    427..448
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             922..1713
                     /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      1806..2394
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2295..2314
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2548..2565
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2724..2751)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2843..2929)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2993..3009
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgtctccatg gcctgacccc ggaccaagtg gtggctatcg agaccggcgc cgctacgggc
       61 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggctctt
      121 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggaacgcc
      181 agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatcgactca
      241 ctatagggcg aattgggtac cgggcccccc ctcgaggtcc tccagctttt gttcccttta
      301 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
      361 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
      421 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccaccg
      481 gtggtctctg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc
      541 caggaccatg gcctgcgaga cgtctagacc agccaggaca gaaatgcctc gacttcgctg
      601 ctacccaagg ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg aacccagtgg
      661 acataagcct gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg
      721 tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt
      781 tatgactgtt tttttggggt acagtctatg cctcgggcat ccaagcagca agcgcgttac
      841 gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca gggcagtcgc
      901 cctaaaacaa agttaaacat tatgagggaa gcggtgatcg ccgaagtatc gactcaacta
      961 tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg
     1021 tacggctccg cagtggatgg cggcctgaag ccacacagtg atattgattt gctggttacg
     1081 gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact
     1141 tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac
     1201 gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag
     1261 cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc
     1321 ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc
     1381 tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg
     1441 aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt
     1501 tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg
     1561 gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga
     1621 caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg
     1681 aaaggcgaga tcaccaaggt agtcggcaaa taaccctcga gccacccatg accaaaatcc
     1741 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1801 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1861 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1921 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     1981 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2041 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2101 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2161 acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc
     2221 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2281 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2341 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2401 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2461 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2521 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2581 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     2641 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc
     2701 caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt
     2761 gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa
     2821 cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac
     2881 agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag
     2941 ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg ttgtaaaacg
     3001 acggccagtc ttaag
//
