LOCUS       sequence-59184-e        2989 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2989)
  AUTHORS   Sakuma T, Hosoi S, Woltjen K, Suzuki KI, Kashiwagi K, Wada H, Ochiai
            H, Miyamoto T, Kawai N, Sasakura Y, Matsuura S, Okada Y, Kawahara A,
            Hayashi S, Yamamoto T
  TITLE     Efficient TALEN construction and evaluation methods for human cell 
            and animal applications.
  JOURNAL   Genes Cells. 2013 Feb 6. doi: 10.1111/gtc.12037.
  PUBMED    23388034
REFERENCE   2  (bases 1 to 2989)
  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: "Genes 
            Cells. 2013 Feb 6. doi: 10.1111/gtc.12037."
COMMENT     Sequence Label: pFUS_A4A
FEATURES             Location/Qualifiers
     source          1..2989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             401..1192
                     /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      1285..1873
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1774..1793
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2027..2044
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2203..2230)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2322..2408)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2462..2839)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLDSH
                     HLVRGQVQGETLKTGRRFTTS"
     primer_bind     2472..2488
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2704..2720
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2730..2749
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2730..2748
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2796..2816)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2796..2814)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2835..2851)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2835..2851)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2848..2870)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2859..2875
                     /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(2883..2889,2890..2907,2908..2913))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2928..2949
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 acggtgcagc ggctgttgcc ggtgctgtgc caggcgagac gtctagacca gccaggacag
       61 aaatgcctcg acttcgctgc tacccaaggt tgccgggtga cgcacaccgt ggaaacggat
      121 gaaggcacga acccagtgga cataagcctg ttcggttcgt aagctgtaat gcaagtagcg
      181 tatgcgctca cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt
      241 ggcggttttc atggcttgtt atgactgttt ttttggggta cagtctatgc ctcgggcatc
      301 caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt
      361 tacgcagcag ggcagtcgcc ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc
      421 cgaagtatcg actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac
      481 gttgctggcc gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga
      541 tattgatttg ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat
      601 caacgacctt ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga
      661 agtcaccatt gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact
      721 gcaatttgga gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat
      781 cgacattgat ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg
      841 tccagcggcg gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa
      901 tgaaacctta acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt
      961 gcttacgttg tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt
     1021 cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc
     1081 tagacaggct tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga
     1141 agaatttgtc cactacgtga aaggcgagat caccaaggta gtcggcaaat aaccctcgag
     1201 ccacccatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     1261 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1321 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1381 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     1441 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1501 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1561 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1621 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagcattg
     1681 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     1741 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     1801 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     1861 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     1921 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     1981 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     2041 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     2101 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     2161 taatacgcgt accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg
     2221 atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct
     2281 ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga
     2341 gcgttcaccg acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt
     2401 tttatttgat gcctggcagt tccctactct cgcgttaacg ctagcatgga tgttttccca
     2461 gtcacgacgt tgtaaaacga cggccagtct taagcgtctc cccctgaacc tgaccccgga
     2521 ccaagtggtg gctatcgaga ccggcgccgc tacagggcgc gtcccattcg ccattcaggc
     2581 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     2641 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     2701 gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta
     2761 ccgggccccc cctcgaggtc ctccagcttt tgttcccttt agtgagggtt aattgcgcgc
     2821 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     2881 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     2941 ctcacattaa ttgcgttgcg ctcactgccc gctttccacc ggtggtctc
//
