LOCUS       sequence-14215-e        2988 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 2988)
  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 2988)
  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_B2
FEATURES             Location/Qualifiers
     source          1..2988
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     206..222
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     232..251
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        232..250
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(298..318)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(298..316)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(337..353)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(337..353)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(350..372)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    361..377
                     /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(385..391,392..409,410..415))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    430..451
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             895..1686
                     /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      1779..2367
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2268..2287
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2521..2538
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2697..2724)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2816..2902)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(2823..2988,1..341))
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLDSH
                     HLVRGQAMRRLRLAVVLQRRDWENIHASVNARVGNCQASNKTKGSVGRLGLSFYLLFVG
                     ERSPE"
     primer_bind     2966..2982
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgtctcatgg cctgaccccg gaccaagtgg tggctatcga gaccggcgcc gctacagggc
       61 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
      121 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
      181 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac
      241 tcactatagg gcgaattggg taccgggccc cccctcgagg tcctccagct tttgttccct
      301 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
      361 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
      421 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
      481 ccggtggtct caggaccatg gcctgacccc ggaccaagtg gtggctatcg agacgtctag
      541 accagccagg acagaaatgc ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca
      601 ccgtggaaac ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg
      661 taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt
      721 ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct
      781 atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg
      841 agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa cattatgagg
      901 gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc
      961 catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg
     1021 aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg
     1081 cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt
     1141 ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca
     1201 gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc
     1261 gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc
     1321 gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta
     1381 tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat
     1441 gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc
     1501 gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc
     1561 gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc
     1621 gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc
     1681 aaataaccct cgagccaccc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac
     1741 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1801 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1861 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1921 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1981 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2041 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2101 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2161 cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2221 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2281 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2341 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2401 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2461 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     2521 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     2581 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     2641 gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa
     2701 ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc
     2761 ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg
     2821 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg
     2881 actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca
     2941 tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaag
//
