LOCUS       sequence-14210-e        3023 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 3023)
  AUTHORS   Cermak T, Doyle EL, Christian M, Wang L, Zhang Y, Schmidt C, Baller 
            JA, Somia NV, Bogdanove AJ, Voytas DF
  TITLE     Efficient design and assembly of custom TALEN and other TAL 
            effector-based constructs for DNA targeting.
  JOURNAL   Nucleic Acids Res. 2011 Jul;39(12):e82. doi: 10.1093/nar/gkr218
  PUBMED    21493687
REFERENCE   2  (bases 1 to 3023)
  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: "Nucleic 
            Acids Res."; date: "2011-07"; volume: "39(12):e82. doi"; pages: " 
            10.1093/nar/gkr21"
COMMENT     Sequence Label: pFUS_B7
FEATURES             Location/Qualifiers
     source          1..3023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     67..83
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     295..311
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     321..340
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        321..339
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(387..407)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(387..405)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(426..442)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(426..442)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(439..461)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    450..466
                     /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(474..480,481..498,499..504))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    519..540
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1019..1810
                     /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      1903..2491
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2392..2411
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2645..2662
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2821..2848)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(join(2940..3023,1..3))
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(2947..3023,1..430))
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLDSH
                     HLVRGQAMRRLRLAVVLQRRDWENIHASVNARVGNCQASNKTKGSVGRLGLSFYLLFVG
                     ERSPE"
ORIGIN
        1 ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac
       61 gacgttgtaa aacgacggcc agtcttaagc gtctcatggc ctgaccccgg accaagtggt
      121 ggctatcgag accggcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
      181 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
      241 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
      301 cgacggccag tgagcgcgcg taatacgact cactataggg cgaattgggt accgggcccc
      361 ccctcgaggt cctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
      421 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
      481 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
      541 attgcgttgc gctcactgcc cgctttccac cggtggtctc cgaaacggtg cagcggctgt
      601 tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct atcgagacgt
      661 ctagaccagc caggacagaa atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg
      721 cacaccgtgg aaacggatga aggcacgaac ccagtggaca taagcctgtt cggttcgtaa
      781 gctgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag
      841 cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttggggtaca
      901 gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt
      961 atggagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacattat
     1021 gagggaagcg gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga
     1081 gcgccatctc gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg
     1141 cctgaagcca cacagtgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac
     1201 aacgcggcga gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga
     1261 gattctccgc gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta
     1321 tccagctaag cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat
     1381 cttcgagcca gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca
     1441 tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga
     1501 tctatttgag gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg
     1561 cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa
     1621 aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca
     1681 gcccgtcata cttgaagcta gacaggctta tcttggacaa gaagaagatc gcttggcctc
     1741 gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt
     1801 cggcaaataa ccctcgagcc acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     1861 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     1921 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     1981 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2041 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2101 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2161 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2221 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2281 cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2341 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2401 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     2461 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     2521 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     2581 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     2641 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     2701 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     2761 cagtgagcgc aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca
     2821 aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg
     2881 cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga
     2941 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
     3001 tccgactgag cctttcgttt tat
//
