LOCUS       sequence-14204-e        3011 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 3011)
  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 3011)
  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_A30B
FEATURES             Location/Qualifiers
     source          1..3011
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     202..218
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     228..247
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        228..246
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(294..314)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(294..312)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(333..349)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(333..349)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(346..368)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    357..373
                     /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(381..387,388..405,406..411))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    426..447
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             918..1709
                     /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      1802..2390
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2291..2310
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2544..2561
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2720..2747)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2839..2925)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2989..3005
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(join(3007..3011,1..337))
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLDSH
                     HLVRGQGDA"
ORIGIN
        1 cgtctccctg accccggacc aagtggtggc tatcgagacc ggcgccgcta cagggcgcgt
       61 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
      121 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
      181 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac
      241 tatagggcga attgggtacc gggccccccc tcgaggtcct ccagcttttg ttccctttag
      301 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
      361 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
      421 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccaccgg
      481 tggtctctgg cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc
      541 aggaccatgg ccgagacgtc tagaccagcc aggacagaaa tgcctcgact tcgctgctac
      601 ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc cagtggacat
      661 aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc aactggtcca
      721 gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg
      781 actgtttttt tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg
      841 tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta
      901 aaacaaagtt aaacattatg agggaagcgg tgatcgccga agtatcgact caactatcag
      961 aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg
     1021 gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg gttacggtga
     1081 ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg
     1141 cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg
     1201 acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca
     1261 atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc
     1321 tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg
     1381 atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact
     1441 cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt
     1501 acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc
     1561 gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag
     1621 aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac tacgtgaaag
     1681 gcgagatcac caaggtagtc ggcaaataac cctcgagcca cccatgacca aaatccctta
     1741 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     1801 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     1861 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     1921 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     1981 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2041 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2101 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2161 cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag
     2221 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2281 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2341 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     2401 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     2461 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     2521 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     2581 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     2641 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc gctagccagg
     2701 aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg
     2761 cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt
     2821 caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat
     2881 aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc
     2941 ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt aaaacgacgg
     3001 ccagtcttaa g
//
