LOCUS       sequence-105133-        3006 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Encodes RVD positions 4-6 in TALEN assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3006)
  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 3006)
  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: pFusX2-25
FEATURES             Location/Qualifiers
     source          1..3006
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     199..215
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(290..310)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(290..308)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(329..345)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(329..345)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(342..364)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    353..369
                     /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(377..383,384..401,402..407))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    422..443
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             913..1704
                     /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      1797..2385
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2286..2305
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2539..2556
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2715..2742)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2834..2920)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2984..3000
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgtctccctg accccggacc aagtggtggc tatcgagacc ggcgccgcta cgggcgcgtc
       61 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg ctcttcgcta
      121 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttggga acgccagggt
      181 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatcg actcactata
      241 gggcgaattg ggtaccgggc cccccctcga ggtcctccag cttttgttcc ctttagtgag
      301 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
      361 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
      421 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc caccggtggt
      481 ctctggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga
      541 ccatggcgag acgtctagac cagccaggac agaaatgcct cgacttcgct gctacccaag
      601 gttgccgggt gacgcacacc gtggaaacgg atgaaggcac gaacccagtg gacataagcc
      661 tgttcggttc gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc
      721 ttgaccgaac gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt
      781 ttttttgggg tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt
      841 cgatgtttga tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca
      901 aagttaaaca ttatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta
      961 gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc
     1021 gcagtggatg gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta
     1081 aggcttgatg aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc
     1141 cctggagaga gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc
     1201 attccgtggc gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac
     1261 attcttgcag gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca
     1321 aaagcaagag aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg
     1381 gttcctgaac aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg
     1441 cccgactggg ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc
     1501 gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg
     1561 ccggcccagt atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa
     1621 gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag
     1681 atcaccaagg tagtcggcaa ataaccctcg agccacccat gaccaaaatc ccttaacgtg
     1741 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1801 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1861 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1921 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     1981 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     2041 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     2101 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     2161 accgaactga gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga
     2221 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2281 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2341 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2401 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2461 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2521 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     2581 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     2641 gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag
     2701 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg
     2761 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat
     2821 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
     2881 gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact
     2941 ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt
     3001 cttaag
//
