LOCUS       sequence-14208-e        3027 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 3027)
  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 3027)
  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_B9
FEATURES             Location/Qualifiers
     source          1..3027
                     /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             1023..1814
                     /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      1907..2495
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2396..2415
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2649..2666
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2825..2852)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(join(2944..3027,1..3))
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(2951..3027,1..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 cgctcgaaac ggtgcagcgg
      601 ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgag
      661 acgtctagac cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt
      721 gacgcacacc gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc
      781 gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac
      841 gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg
      901 tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga
      961 tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca
     1021 ttatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca
     1081 tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg
     1141 gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg
     1201 aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga
     1261 gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc
     1321 gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag
     1381 gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag
     1441 aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac
     1501 aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg
     1561 ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg
     1621 gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt
     1681 atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg
     1741 cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg
     1801 tagtcggcaa ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt
     1861 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1921 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1981 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2041 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2101 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2161 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2221 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     2281 gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2341 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2401 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2461 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2521 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     2581 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     2641 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     2701 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     2761 cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa
     2821 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg
     2881 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg
     2941 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
     3001 gtcttccgac tgagcctttc gttttat
//
