LOCUS       sequence-59182-e        2993 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 2993)
  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 2993)
  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_A3A
FEATURES             Location/Qualifiers
     source          1..2993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             405..1196
                     /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      1289..1877
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1778..1797
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2031..2048
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2207..2234)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2326..2412)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2466..2843)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLDSH
                     HLVRGQVQGETLKTGRRFTTS"
     primer_bind     2476..2492
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2708..2724
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2734..2753
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2734..2752
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2800..2820)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2800..2818)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2839..2855)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2839..2855)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2852..2874)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2863..2879
                     /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(2887..2893,2894..2911,2912..2917))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2932..2953
                     /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 caggaccacg agacgtctag accagccagg
       61 acagaaatgc ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac
      121 ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt
      181 agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg
      241 cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg
      301 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg
      361 atgttacgca gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga
      421 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac
      481 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca
      541 gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt
      601 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg
      661 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg
      721 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca
      781 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg
      841 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc
      901 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg
      961 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg
     1021 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg
     1081 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt
     1141 tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct
     1201 cgagccaccc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag
     1261 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     1321 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1381 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     1441 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1501 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     1561 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     1621 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     1681 attgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     1741 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     1801 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     1861 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     1921 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     1981 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     2041 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     2101 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     2161 caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt
     2221 caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca
     2281 ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag
     2341 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt
     2401 tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca tggatgtttt
     2461 cccagtcacg acgttgtaaa acgacggcca gtcttaagcg tctccccctg aacctgaccc
     2521 cggaccaagt ggtggctatc gagaccggcg ccgctacagg gcgcgtccca ttcgccattc
     2581 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     2641 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     2701 cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg
     2761 ggtaccgggc cccccctcga ggtcctccag cttttgttcc ctttagtgag ggttaattgc
     2821 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     2881 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     2941 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc caccggtggt ctc
//
