LOCUS       sequence-105345-        3008 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Encodes RVD positions 10-12 in TALEN assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3008)
  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 3008)
  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: pFusX4-4
FEATURES             Location/Qualifiers
     source          1..3008
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     200..216
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(291..311)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(291..309)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(330..346)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(330..346)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(343..365)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    354..370
                     /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(378..384,385..402,403..408))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    423..444
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             915..1706
                     /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      1799..2387
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2288..2307
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2541..2558
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2717..2744)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2836..2922)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2986..3002
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgtctcccct gaccccggac caagtggtgg ctatcgagac cggcgccgct acgggcgcgt
       61 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gctcttcgct
      121 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg aacgccaggg
      181 ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaatc gactcactat
      241 agggcgaatt gggtaccggg ccccccctcg aggtcctcca gcttttgttc cctttagtga
      301 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
      361 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
      421 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccaccggtgg
      481 tctctggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg
      541 accatggccg agacgtctag accagccagg acagaaatgc ctcgacttcg ctgctaccca
      601 aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag tggacataag
      661 cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa
      721 ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact
      781 gtttttttgg ggtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg
      841 gtcgatgttt gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa
      901 caaagttaaa cattatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg
      961 tagttggcgt catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct
     1021 ccgcagtgga tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg
     1081 taaggcttga tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt
     1141 cccctggaga gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca
     1201 tcattccgtg gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg
     1261 acattcttgc aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga
     1321 caaaagcaag agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc
     1381 cggttcctga acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc
     1441 cgcccgactg ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca
     1501 gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc
     1561 tgccggccca gtatcagccc gtcatacttg aagctagaca ggcttatctt ggacaagaag
     1621 aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt tgtccactac gtgaaaggcg
     1681 agatcaccaa ggtagtcggc aaataaccct cgagccaccc atgaccaaaa tcccttaacg
     1741 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     1801 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     1861 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     1921 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     1981 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2041 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2101 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2161 acaccgaact gagataccta cagcgtgagc attgagaaag cgccacgctt cccgaaggga
     2221 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2281 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2341 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2401 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     2461 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     2521 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     2581 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     2641 ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag
     2701 agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct
     2761 ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa
     2821 atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
     2881 acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta
     2941 ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca
     3001 gtcttaag
//
