LOCUS       sequence-59179-e        2997 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 2997)
  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 2997)
  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_A1A
FEATURES             Location/Qualifiers
     source          1..2997
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             409..1200
                     /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      1293..1881
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1782..1801
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2035..2052
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2211..2238)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2330..2416)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2470..2847)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLDSH
                     HLVRGQVQGETLKTGRRFTTS"
     primer_bind     2480..2496
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2712..2728
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2738..2757
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2738..2756
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2804..2824)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2804..2822)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2843..2859)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2843..2859)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2856..2878)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2867..2883
                     /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(2891..2897,2898..2915,2916..2921))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2936..2957
                     /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 gccgagacgt ctagaccagc
       61 caggacagaa atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg
      121 aaacggatga aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc
      181 aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac
      241 ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct
      301 cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc
      361 aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacattat gagggaagcg
      421 gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc
      481 gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca
      541 cacagtgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga
      601 gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc
      661 gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag
      721 cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca
      781 gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc
      841 ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag
      901 gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga
      961 aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg
     1021 aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata
     1081 cttgaagcta gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat
     1141 cagttggaag aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa
     1201 ccctcgagcc acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg
     1261 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     1321 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     1381 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     1441 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     1501 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     1561 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     1621 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     1681 gagcattgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     1741 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     1801 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     1861 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     1921 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     1981 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     2041 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     2101 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     2161 aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat
     2221 ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc
     2281 gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac
     2341 tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag
     2401 cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg
     2461 ttttcccagt cacgacgttg taaaacgacg gccagtctta agcgtctccc cctgaacctg
     2521 accccggacc aagtggtggc tatcgagacc ggcgccgcta cagggcgcgt cccattcgcc
     2581 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     2641 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     2701 gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga
     2761 attgggtacc gggccccccc tcgaggtcct ccagcttttg ttccctttag tgagggttaa
     2821 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     2881 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     2941 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccaccgg tggtctc
//
