LOCUS       sequence-59180-e        3001 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 3001)
  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 3001)
  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_A2A
FEATURES             Location/Qualifiers
     source          1..3001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             413..1204
                     /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      1297..1885
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1786..1805
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2039..2056
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2215..2242)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2334..2420)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2474..2851)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLDSH
                     HLVRGQVQGETLKTGRRFTTS"
     primer_bind     2484..2500
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2716..2732
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2742..2761
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2742..2760
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2808..2828)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2808..2826)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2847..2863)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2847..2863)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2860..2882)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2871..2887
                     /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(2895..2901,2902..2919,2920..2925))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2940..2961
                     /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 caggaccatg gcctgacgag acgtctagac
       61 cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc
      121 gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta
      181 atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg
      241 taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat
      301 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag
      361 cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca ttatgaggga
      421 agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca
      481 tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa
      541 gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg
      601 gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct
      661 ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc
      721 taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga
      781 gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt
      841 tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt
      901 tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga
      961 gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc
     1021 gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt
     1081 catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc
     1141 agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa
     1201 ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     1261 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     1321 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     1381 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     1441 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     1501 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     1561 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     1621 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     1681 gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     1741 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     1801 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     1861 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     1921 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     1981 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     2041 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     2101 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     2161 gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg
     2221 ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct
     2281 gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc
     2341 ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac
     2401 tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg
     2461 gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagcgtc tccccctgaa
     2521 cctgaccccg gaccaagtgg tggctatcga gaccggcgcc gctacagggc gcgtcccatt
     2581 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     2641 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     2701 cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg
     2761 gcgaattggg taccgggccc cccctcgagg tcctccagct tttgttccct ttagtgaggg
     2821 ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     2881 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     2941 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca ccggtggtct
     3001 c
//
