LOCUS       sequence-190036-        6778 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TALE targeted to lac operator with one TEV cut site in its
            N-terminal domain and a second in the C-terminal domain, 
            anhydrotetracycline inducible TEV protease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6778)
  AUTHORS   Copeland MF, Politz MC, Johnson CB, Markley AL, Pfleger BF
  TITLE     A transcription activator-like effector (TALE) induction system 
            mediated by proteolysis.
  JOURNAL   Nat Chem Biol. 2016 Apr;12(4):254-60. doi: 10.1038/nchembio.2021. 
            Epub 2016 Feb 8.
  PUBMED    26854666
REFERENCE   2  (bases 1 to 6778)
  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"; doi: 
            "10.1038/nchembio.2021"; journalName: "Nat Chem Biol"; date: 
            "2016-04"; volume: "12"; issue: "4"; pages: "254-60"
COMMENT     Sequence Label: p5A7-T
FEATURES             Location/Qualifiers
     source          1..6778
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..21
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             22..729
                     /codon_start=1
                     /product="tobacco etch virus protease (Kapust et al., 
                     2001)"
                     /label=TEV protease
                     /translation="GESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNK
                     HLFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQ
                     REERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFI
                     VGIHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPE
                     EPFQPVKEATQLMN"
     terminator      816..852
                     /label=soxR terminator
                     /note="bidirectional E. coli soxR transcription terminator"
     misc_feature    903..924
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        925..959
                     /label=BBa_J23102
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.86"
     CDS             1003..1020
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1438..1458
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3307..3327
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3586..3609
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      3647..3678
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(3695..4510)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MNHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4418..4437
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(4600..5265)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRRT"
     terminator      complement(5925..5996)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(6041..6667)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     6572..6591
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        6683..6738
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    6719..6737
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 catcatcatc atcatcatca tggagaaagc ttgtttaagg ggccgcgtga ttacaacccg
       61 atatcgagca ccatttgtca tttgacgaat gaatctgatg ggcacacaac atcgttgtat
      121 ggtattggat ttggtccctt catcattaca aacaagcact tgtttagaag aaataatgga
      181 acactgttgg tccaatcact acatggtgta ttcaaggtca agaacaccac gactttgcaa
      241 caacacctca ttgatgggag ggacatgata attattcgca tgcctaagga tttcccacca
      301 tttcctcaaa agctgaaatt tagagagcca caaagggaag agcgcatatg tcttgtgaca
      361 accaacttcc aaactaagag catgtctagc atggtgtcag acactagttg cacattccct
      421 tcatctgatg gcatattctg gaagcattgg attcaaacca aggatgggca gtgtggcagt
      481 ccattagtat caactagaga tgggttcatt gttggtatac actcagcatc gaatttcacc
      541 aacacaaaca attatttcac aagcgtgccg aaaaacttca tggaattgtt gacaaatcag
      601 gaggcgcagc agtgggttag tggttggcga ttaaatgctg actcagtatt gtgggggggc
      661 cataaagttt tcatggtgaa acctgaagag ccttttcagc cagttaagga agcgactcaa
      721 ctcatgaatc gtcgtcgccg tcgctaagtc catcagcttg tccagcaggg ttgtccacgg
      781 gccgagcgaa gcgagccagc cggtggccgc ggaacaaaac taaagcgcca caagggcgct
      841 ttagtttgtt ttcagtccag ttacgctgga gtctgaggct cgtcctgaat gatatcaagc
      901 ttgaattcgc ggccgcttct agagttgaca gctagctcag tcctaggtac tgtgctagct
      961 actagatatc gctcaatact gaccaaggag gtatctacaa tgcatcacca tcatcaccat
     1021 gttgatctgc gtaccctggg ttatagccag cagcagcaag aaaaaatcaa accgaaagtt
     1081 cgtagcaccg ttgcacagca tcatgaagca ctggttggtc atggttttac ccatgcacat
     1141 attgttgcac tgagccagca tccggcagca ctgggcaccg ttgcagttaa atatcaggat
     1201 atgattgcag cactgccgga agcaacccat gaagcaattg ttggtgttgg taaacagtgg
     1261 tcaggtgcac gtgccctgga agcactgctg accgttgccg gtgaactgcg tggtccgcct
     1321 ctgcagctgg ataccggtca gctgctgaaa attgcaaaac gtggtggtgt taccgcagtt
     1381 gaagcagttc atgcatggcg taatgcactg accggtgcac cgctgaatgg ttctggcgaa
     1441 aacctgtact tccagtctgg ctcgggtctg acaccggaac aggttgttgc aattgccagc
     1501 aatggtggtg gcaaacaggc actggaaacc gttcagcgtc tgctgccggt tctgtgtcag
     1561 gcacatggtc tgacccctga acaggtggtg gccattgcaa gcaataatgg cggtaaacaa
     1621 gccctggaaa cagtgcagcg cctgttaccg gtgctgtgcc aggcccatgg cttaactccg
     1681 gaacaggtgg tagcgatcgc atcaaatggc ggaggtaaac aggccttaga aaccgtacag
     1741 cgcttactgc cggtgttatg ccaggcgcac ggcctgacgc cagaacaggt agtggcaatc
     1801 gcctcaaata atggtggaaa acaggcgtta gagacagtcc agcgcctgct gcctgtatta
     1861 tgtcaagccc atggcctgac cccagagcaa gttgttgcga ttgcaagtaa cattgggggt
     1921 aaacaggcac ttgagacagt tcaacgttta ctgcctgtac tgtgccaagc tcacggtctg
     1981 actccggaac aagtcgtcgc gattgcgagt aataacggtg gcaaacaagc attagaaacg
     2041 gtgcaacgcc tgctgccagt tctttgccag gctcacggtt taacccctga gcaggttgta
     2101 gctattgcga gtcatgatgg tggtaagcag gcgttggaaa ctgtgcaaag actgctgccc
     2161 gtgttgtgcc aagcacatgg tttaacccca gaacaagtcg tagcaatcgc aagcaatcat
     2221 ggtggcaagc aagcgcttga aacagtacag cgtttattac cggtactttg tcaggcccac
     2281 ggtcttacac cagaacaagt tgtggccata gccagtaaca atggcggaaa gcaggctctg
     2341 gaaacggtac aacgtctgtt acctgttctg tgtcaagcgc acggattaac acctgaacaa
     2401 gtagttgcca ttgcgtcaaa tatcggaggc aagcaggcct tggagacagt gcagagatta
     2461 ctgccagtgt tgtgtcaggc tcatggcctt acacccgagc aggtcgtggc aattgcatct
     2521 aatggtggcg gtaagcaagc tttagagact gttcagagac tgcttcctgt cctgtgccag
     2581 gcacacggac ttacgcctga gcaagtggtt gcaatcgcct ctaacatcgg tggtaagcaa
     2641 gcactggaaa ctgtccaacg cttacttccg gtgctttgtc aagcacacgg cttaacgcca
     2701 gagcaggtcg tcgccatagc cagcaatatc ggtggtaaac aggcccttga aacggtccaa
     2761 agacttctgc cggtcctttg ccaagcgcat gggctgacac ctgagcaggt agtcgcgatt
     2821 gcctcacatg acggtgggaa gcaggcatta gaaacagttc aaagattatt accagtcctg
     2881 tgtcaggcgc atgggttaac cccagagcag gtagttgcaa tagcatccaa cattggcgga
     2941 aaacaagcgt tggaaacggt tcagcggtta ttgcctgttt tgtgccaggc gcatggtttg
     3001 acacccgagc aagtggtagc catagcctca aatatagggg gtaaacaagc tttggagaca
     3061 gtacaacggc tgcttccagt tttatgtcag gcccatggat tgacgcctga acaagttgtc
     3121 gctatcgcaa gtaatggggg tggtaaacaa gcgcttgaaa ccgttcaacg ccttctgcct
     3181 gtgctttgtc aggcacatgg attaacaccc gaacaggttg tcgcgatagc ttcaaacggt
     3241 ggtggtcgtc cggcactgga aagcattgtt gcacagctga gccgtccgga tccggcaggt
     3301 agcggtgaaa acctgtattt ccaatctggc agcggcctgg cagcactgac caatgatcat
     3361 ctggttgcac tggcatgtct gggtggtcgc cctgccctgg atgcagttaa aaaaggtctg
     3421 ccgcatgcac cggcactgat taaacgtacc aatcgtcgta ttccggaacg taccagccat
     3481 cgtgttgcag atcatgcaca ggttgttcgt gttctgggtt tttttcagtg tcatagccat
     3541 ccggcacagg catttgatga tgcaatgacc cagtttggta tgagcgatta taaagatgat
     3601 gatgataaat aaggtaccct aatcatacct gacctccata gcagaaagtc aaaaacctcc
     3661 gaccggaggc ttttgacttg agggggatcc gccctcagaa aaactcatcg agcatcaagt
     3721 gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa agccgtttct
     3781 gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt
     3841 ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa
     3901 ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagct
     3961 tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac
     4021 tcgcaccaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat
     4081 cgccgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca
     4141 gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt
     4201 tccctgggat cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga
     4261 tggtcggaag aggcataaat tccgtcagcc agtttagcct gaccatctca tctgtaacat
     4321 cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat
     4381 acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat
     4441 ataaatcagc atccatgttg gaatttaatc gcggcctcga gcaagacgtt tcccgttgaa
     4501 tatggttcat tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg
     4561 gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtacggc gcggcagcgt
     4621 gacccgtgtc ggcggctcca acggctcgcc atcgtccaga aaacacggct catcgggcat
     4681 cggcaggcgc tgctgcccgc gtcgttccca ttcctccgtt tcggtcaagg ctggcaggtc
     4741 tggttccatg cccggaatgc cgggctggct gggcggctcc tcgccggggc cggtcggtag
     4801 ttgctgctcg cccggataca gggtcgggat gcggcgcagg tcgccatgcc ccaacagcga
     4861 ttcgtcctgg tcgtcgtgat caaccaccac ggcggcactg aacaccgaca ggcgcaactg
     4921 gtcgcggggc tggccccacg ccacgcggtc attgaccacg taggccgaca cggtgccggg
     4981 gccgttgagc ttcacgacgg agatccagcg ctcggccacc aagtccttga ctgcgtattg
     5041 gaccgtccgc aaagaacgtc cgatgagctt ggaaagtgtc ttctggctga ccaccacggc
     5101 gttctggtgg cccatctgcg ccacgaggtg atgcagcagc attgccgccg tgggtttcct
     5161 cgcaataagc ccggcccacg cctcatgcgc tttgcgttcc gtttgcaccc agtgaccggg
     5221 cttgttcttg gcttgaatgc cgatttctct ggactgcgtg gccatgctta tctccatgcg
     5281 gtaggggtgc cgcacggttg cggcaccatg cgcaatcagc tgcaactttt cggcagcgcg
     5341 acaacaatta tgcgttgcgt aaaagtggca gtcaattaca gattttcttt aacctacgca
     5401 atgagctatt gcggggggtg ccgcaatgag ctgttgcgta cccccctttt ttaagttgtt
     5461 gatttttaag tctttcgcat ttcgccctat atctagttct ttggtgccca aagaagggca
     5521 cccctgcggg gttcccccac gccttcggcg cggctccccc tccggcaaaa agtggcccct
     5581 ccggggcttg ttgatcgact gcgcggcctt cggccttgcc caaggtggcg ctgccccctt
     5641 ggaacccccg cactcgccgc cgtgaggctc ggggggcagg cgggcgggct tcgcccttcg
     5701 actgccccca ctcgcatggg cttgggtcgt tccaggcgcg tcaaggccaa gccgctgcgc
     5761 ggtcgctgcg cgagccttga cccgccttcc acttggtgtc caaccggcaa gcgaagcgcg
     5821 caggccgcag gccggaggct tttccccaga gaaaattaaa aaaattgatg gggcaaggcc
     5881 gcaggccgcg cagttggagc cggtgggtat gtggtcgaag gctgggtagc gttcaccgac
     5941 aaacaacaga taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc
     6001 ctggctctag tagtgatcta cactagcact atcagtgtta ttaagaccca ctttcacatt
     6061 taagttgttt ttctaatccg catatgatca attcaaggcc gaataagaag gctggctctg
     6121 caccttggtg atcaaataat tcgatagctt gtcgtaataa tggcggcata ctatcagtag
     6181 taggtgtttc cctttcttct ttagcgactt gatgctcttg atcttccaat acgcaaccta
     6241 aagtaaaatg ccccacagcg ctgagtgcat ataatgcatt ctctagtgaa aaaccttgtt
     6301 ggcataaaaa ggctaattga ttttcgagag tttcatactg tttttctgta ggccgtgtac
     6361 ctaaatgtac ttttgctcca tcgcgatgac ttagtaaagc acatctaaaa cttttagcgt
     6421 tattacgtaa aaaatcttgc cagctttccc cttctaaagg gcaaaagtga gtatggtgcc
     6481 tatctaacat ctcaatggct aaggcgtcga gcaaagcccg cttatttttt acatgccaat
     6541 acaatgtagg ctgctctaca cctagcttct gggcgagttt acgggttgtt aaaccttcga
     6601 ttccgacctc attaagcagc tctaatgcgc tgttaatcac tttactttta tctaatctag
     6661 acatcattaa ttcctaattt ttgttgacac tctatcgttg atagagttat tttaccactc
     6721 cctatcagtg atagagaaaa gaattcaaaa gatcttttaa gaaggagata tacatatg
//
