LOCUS       sequence-104963-        3013 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Encodes RVD positions 1-3 in TALEN assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3013)
  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 3013)
  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: pFusX1-4
FEATURES             Location/Qualifiers
     source          1..3013
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     201..217
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(292..312)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(292..310)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(331..347)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(331..347)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(344..366)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    355..371
                     /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(379..385,386..403,404..409))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    424..445
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             920..1711
                     /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      1804..2392
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2293..2312
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2546..2563
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2722..2749)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2841..2927)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2991..3007
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgtctccccc tgaccccgga ccaagtggtg gctatcgaga ccggcgccgc tacgggcgcg
       61 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggctcttcgc
      121 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gaacgccagg
      181 gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat cgactcacta
      241 tagggcgaat tgggtaccgg gccccccctc gaggtcctcc agcttttgtt ccctttagtg
      301 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
      361 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
      421 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccaccggtg
      481 gtctctggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag
      541 gaccatggcc tgacgagacg tctagaccag ccaggacaga aatgcctcga cttcgctgct
      601 acccaaggtt gccgggtgac gcacaccgtg gaaacggatg aaggcacgaa cccagtggac
      661 ataagcctgt tcggttcgta agctgtaatg caagtagcgt atgcgctcac gcaactggtc
      721 cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca tggcttgtta
      781 tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc
      841 cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg gcagtcgccc
      901 taaaacaaag ttaaacatta tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc
      961 agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg tacatttgta
     1021 cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc tggttacggt
     1081 gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt tggaaacttc
     1141 ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg ttgtgcacga
     1201 cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag aatggcagcg
     1261 caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc tggctatctt
     1321 gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg aggaactctt
     1381 tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa cgctatggaa
     1441 ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg
     1501 gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga
     1561 gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt atcttggaca
     1621 agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa
     1681 aggcgagatc accaaggtag tcggcaaata accctcgagc cacccatgac caaaatccct
     1741 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     1801 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     1861 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     1921 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     1981 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2041 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2101 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2161 gacctacacc gaactgagat acctacagcg tgagcattga gaaagcgcca cgcttcccga
     2221 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2281 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2341 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2401 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     2461 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     2521 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     2581 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     2641 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca
     2701 ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga
     2761 tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg
     2821 ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag
     2881 ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt
     2941 ccctactctc gcgttaacgc tagcatggat gttttcccag tcacgacgtt gtaaaacgac
     3001 ggccagtctt aag
//
