LOCUS       sequence-75845-e        2935 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Intermediate array vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2935)
  AUTHORS   Sakuma T, Ochiai H, Kaneko T, Mashimo T, Tokumasu D, Sakane Y, 
            Suzuki K, Miyamoto T, Sakamoto N, Matsuura S, Yamamoto T
  TITLE     Repeating pattern of non-RVD variations in DNA-binding modules 
            enhances TALEN activity.
  JOURNAL   Sci Rep. 2013 Nov 29;3:3379. doi: 10.1038/srep03379.
  PUBMED    24287550
REFERENCE   2  (bases 1 to 2935)
  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: "Sci Rep."; 
            date: "2013-11-29"; volume: "3:3379. doi"; pages: " 
            10.1038/srep03379"
COMMENT     Sequence Label: pFUS2_a3b
FEATURES             Location/Qualifiers
     source          1..2935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             367..1158
                     /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      1251..1839
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1740..1759
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1993..2010
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2169..2196)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2288..2374)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2428..2778)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLGVS
                     ETLKTGRRFTTS"
     primer_bind     2438..2454
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2643..2659
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2669..2688
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2669..2687
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2735..2755)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2735..2753)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2774..2790)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2774..2790)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2787..2809)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2798..2814
                     /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(2822..2828,2829..2846,2847..2852))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2867..2888
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cgagacgtct agaccagcca ggacagaaat gcctcgactt cgctgctacc caaggttgcc
       61 gggtgacgca caccgtggaa acggatgaag gcacgaaccc agtggacata agcctgttcg
      121 gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc
      181 gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt
      241 ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt
      301 ttgatgttat ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta
      361 aacattatga gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc
      421 gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg
      481 gatggcggcc tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt
      541 gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga
      601 gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg
      661 tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt
      721 gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca
      781 agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct
      841 gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac
      901 tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta
      961 accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc
     1021 cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc
     1081 ttggcctcgc gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc
     1141 aaggtagtcg gcaaataacc ctcgagccac ccatgaccaa aatcccttaa cgtgagttac
     1201 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1261 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1321 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     1381 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     1441 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     1501 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     1561 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     1621 ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg
     1681 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     1741 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     1801 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     1861 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     1921 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     1981 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     2041 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     2101 aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta
     2161 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt
     2221 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc
     2281 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     2341 cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg
     2401 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaag
     2461 cgtctcactg accccgagac cggcgccgct acagggcgcg tcccattcgc cattcaggct
     2521 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     2581 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     2641 ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac
     2701 cgggcccccc ctcgaggtcc tccagctttt gttcccttta gtgagggtta attgcgcgct
     2761 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     2821 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     2881 tcacattaat tgcgttgcgc tcactgcccg ctttccaccg gtggtctcac ctgac
//
