LOCUS       sequence-201851-        6700 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TALE targeted to lac operator without TEV cut sites, 
            anhydrotetracycline inducible TEV protease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6700)
  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 6700)
  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: p5A0-T
FEATURES             Location/Qualifiers
     source          1..6700
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..22
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        23..57
                     /label=BBa_J23102
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.86"
     CDS             101..118
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2606..2629
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      2667..2698
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(2715..3530)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3438..3457
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(3620..4285)
                     /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(4945..5016)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(5061..5687)
                     /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     5592..5611
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        5703..5758
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    5739..5757
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             5799..5819
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             5820..6527
                     /codon_start=1
                     /product="tobacco etch virus protease (Kapust et al., 
                     2001)"
                     /label=TEV protease
                     /translation="GESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNK
                     HLFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQ
                     REERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFI
                     VGIHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPE
                     EPFQPVKEATQLMN"
     terminator      6614..6650
                     /label=soxR terminator
                     /note="bidirectional E. coli soxR transcription terminator"
ORIGIN
        1 gaattcgcgg ccgcttctag agttgacagc tagctcagtc ctaggtactg tgctagctac
       61 tagatatcgc tcaatactga ccaaggaggt atctacaatg catcaccatc atcaccatgt
      121 tgatctgcgt accctgggtt atagccagca gcagcaagaa aaaatcaaac cgaaagttcg
      181 tagcaccgtt gcacagcatc atgaagcact ggttggtcat ggttttaccc atgcacatat
      241 tgttgcactg agccagcatc cggcagcact gggcaccgtt gcagttaaat atcaggatat
      301 gattgcagca ctgccggaag caacccatga agcaattgtt ggtgttggta aacagtggtc
      361 aggtgcacgt gccctggaag cactgctgac cgttgccggt gaactgcgtg gtccgcctct
      421 gcagctggat accggtcagc tgctgaaaat tgcaaaacgt ggtggtgtta ccgcagttga
      481 agcagttcat gcatggcgta atgcactgac cggtgcaccg ctgaatctga caccggaaca
      541 ggttgttgca attgccagca atggtggtgg caaacaggca ctggaaaccg ttcagcgtct
      601 gctgccggtt ctgtgtcagg cacatggtct gacccctgaa caggtggtgg ccattgcaag
      661 caataatggc ggtaaacaag ccctggaaac agtgcagcgc ctgttaccgg tgctgtgcca
      721 ggcccatggc ttaactccgg aacaggtggt agcgatcgca tcaaatggcg gaggtaaaca
      781 ggccttagaa accgtacagc gcttactgcc ggtgttatgc caggcgcacg gcctgacgcc
      841 agaacaggta gtggcaatcg cctcaaataa tggtggaaaa caggcgttag agacagtcca
      901 gcgcctgctg cctgtattat gtcaagccca tggcctgacc ccagagcaag ttgttgcgat
      961 tgcaagtaac attgggggta aacaggcact tgagacagtt caacgtttac tgcctgtact
     1021 gtgccaagct cacggtctga ctccggaaca agtcgtcgcg attgcgagta ataacggtgg
     1081 caaacaagca ttagaaacgg tgcaacgcct gctgccagtt ctttgccagg ctcacggttt
     1141 aacccctgag caggttgtag ctattgcgag tcatgatggt ggtaagcagg cgttggaaac
     1201 tgtgcaaaga ctgctgcccg tgttgtgcca agcacatggt ttaaccccag aacaagtcgt
     1261 agcaatcgca agcaatcatg gtggcaagca agcgcttgaa acagtacagc gtttattacc
     1321 ggtactttgt caggcccacg gtcttacacc agaacaagtt gtggccatag ccagtaacaa
     1381 tggcggaaag caggctctgg aaacggtaca acgtctgtta cctgttctgt gtcaagcgca
     1441 cggattaaca cctgaacaag tagttgccat tgcgtcaaat atcggaggca agcaggcctt
     1501 ggagacagtg cagagattac tgccagtgtt gtgtcaggct catggcctta cacccgagca
     1561 ggtcgtggca attgcatcta atggtggcgg taagcaagct ttagagactg ttcagagact
     1621 gcttcctgtc ctgtgccagg cacacggact tacgcctgag caagtggttg caatcgcctc
     1681 taacatcggt ggtaagcaag cactggaaac tgtccaacgc ttacttccgg tgctttgtca
     1741 agcacacggc ttaacgccag agcaggtcgt cgccatagcc agcaatatcg gtggtaaaca
     1801 ggcccttgaa acggtccaaa gacttctgcc ggtcctttgc caagcgcatg ggctgacacc
     1861 tgagcaggta gtcgcgattg cctcacatga cggtgggaag caggcattag aaacagttca
     1921 aagattatta ccagtcctgt gtcaggcgca tgggttaacc ccagagcagg tagttgcaat
     1981 agcatccaac attggcggaa aacaagcgtt ggaaacggtt cagcggttat tgcctgtttt
     2041 gtgccaggcg catggtttga cacccgagca agtggtagcc atagcctcaa atataggggg
     2101 taaacaagct ttggagacag tacaacggct gcttccagtt ttatgtcagg cccatggatt
     2161 gacgcctgaa caagttgtcg ctatcgcaag taatgggggt ggtaaacaag cgcttgaaac
     2221 cgttcaacgc cttctgcctg tgctttgtca ggcacatgga ttaacacccg aacaggttgt
     2281 cgcgatagct tcaaacggtg gtggtcgtcc ggcactggaa agcattgttg cacagctgag
     2341 ccgtccggat ccggcactgg cagcactgac caatgatcat ctggttgcac tggcatgtct
     2401 gggtggtcgc cctgccctgg atgcagttaa aaaaggtctg ccgcatgcac cggcactgat
     2461 taaacgtacc aatcgtcgta ttccggaacg taccagccat cgtgttgcag atcatgcaca
     2521 ggttgttcgt gttctgggtt tttttcagtg tcatagccat ccggcacagg catttgatga
     2581 tgcaatgacc cagtttggta tgagcgatta taaagatgat gatgataaat aaggtaccct
     2641 aatcatacct gacctccata gcagaaagtc aaaaacctcc gaccggaggc ttttgacttg
     2701 agggggatcc gccctcagaa aaactcatcg agcatcaagt gaaactgcaa tttattcata
     2761 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
     2821 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
     2881 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
     2941 ccatgagtga cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact
     3001 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcaccaac caaaccgtta
     3061 ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgccgttaaa aggacaatta
     3121 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     3181 cctgaatcag gatattcttc taatacctgg aatgctgttt tccctgggat cgcagtggtg
     3241 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     3301 tccgtcagcc agtttagcct gaccatctca tctgtaacat cattggcaac gctacctttg
     3361 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca
     3421 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     3481 gaatttaatc gcggcctcga gcaagacgtt tcccgttgaa tatggctcat tttagcttcc
     3541 ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta
     3601 tggtgaaagt tggaacctct tacgtacggc gcggcagcgt gacccgtgtc ggcggctcca
     3661 acggctcgcc atcgtccaga aaacacggct catcgggcat cggcaggcgc tgctgcccgc
     3721 gtcgttccca ttcctccgtt tcggtcaagg ctggcaggtc tggttccatg cccggaatgc
     3781 cgggctggct gggcggctcc tcgccggggc cggtcggtag ttgctgctcg cccggataca
     3841 gggtcgggat gcggcgcagg tcgccatgcc ccaacagcga ttcgtcctgg tcgtcgtgat
     3901 caaccaccac ggcggcactg aacaccgaca ggcgcaactg gtcgcggggc tggccccacg
     3961 ccacgcggtc attgaccacg taggccgaca cggtgccggg gccgttgagc ttcacgacgg
     4021 agatccagcg ctcggccacc aagtccttga ctgcgtattg gaccgtccgc aaagaacgtc
     4081 cgatgagctt ggaaagtgtc ttctggctga ccaccacggc gttctggtgg cccatctgcg
     4141 ccacgaggtg atgcagcagc attgccgccg tgggtttcct cgcaataagc ccggcccacg
     4201 cctcatgcgc tttgcgttcc gtttgcaccc agtgaccggg cttgttcttg gcttgaatgc
     4261 cgatttctct ggactgcgtg gccatgctta tctccatgcg gtaggggtgc cgcacggttg
     4321 cggcaccatg cgcaatcagc tgcaactttt cggcagcgcg acaacaatta tgcgttgcgt
     4381 aaaagtggca gtcaattaca gattttcttt aacctacgca atgagctatt gcggggggtg
     4441 ccgcaatgag ctgttgcgta cccccctttt ttaagttgtt gatttttaag tctttcgcat
     4501 ttcgccctat atctagttct ttggtgccca aagaagggca cccctgcggg gttcccccac
     4561 gccttcggcg cggctccccc tccggcaaaa agtggcccct ccggggcttg ttgatcgact
     4621 gcgcggcctt cggccttgcc caaggtggcg ctgccccctt ggaacccccg cactcgccgc
     4681 cgtgaggctc ggggggcagg cgggcgggct tcgcccttcg actgccccca ctcgcatagg
     4741 cttgggtcgt tccaggcgcg tcaaggccaa gccgctgcgc ggtcgctgcg cgagccttga
     4801 cccgccttcc acttggtgtc caaccggcaa gcgaagcgcg caggccgcag gccggaggct
     4861 tttccccaga gaaaattaaa aaaattgatg gggcaaggcc gcaggccgcg cagttggagc
     4921 cggtgggtat gtggtcgaag gctgggtagc gttcaccgac aaacaacaga taaaacgaaa
     4981 ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggctctag tagtgatcta
     5041 cactagcact atcagtgtta ttaagaccca ctttcacatt taagttgttt ttctaatccg
     5101 catatgatca attcaaggcc gaataagaag gctggctctg caccttggtg atcaaataat
     5161 tcgatagctt gtcgtaataa tggcggcata ctatcagtag taggtgtttc cctttcttct
     5221 ttagcgactt gatgctcttg atcttccaat acgcaaccta aagtaaaatg ccccacagcg
     5281 ctgagtgcat ataatgcatt ctctagtgaa aaaccttgtt ggcataaaaa ggctaattga
     5341 ttttcgagag tttcatactg tttttctgta ggccgtgtac ctaaatgtac ttttgctcca
     5401 tcgcgatgac ttagtaaagc acatctaaaa cttttagcgt tattacgtaa aaaatcttgc
     5461 cagctttccc cttctaaagg gcaaaagtga gtatggtgcc tatctaacat ctcaatggct
     5521 aaggcgtcga gcaaagcccg cttatttttt acatgccaat acaatgtagg ctgctctaca
     5581 cctagcttct gggcgagttt acgggttgtt aaaccttcga ttccgacctc attaagcagc
     5641 tctaatgcgc tgttaatcac tttactttta tctaatctag acatcattaa ttcctaattt
     5701 ttgttgacac tctatcgttg atagagttat tttaccactc cctatcagtg atagagaaaa
     5761 gaattcaaaa gatcttttaa gaaggagata tacatatgca tcatcatcat catcatcatg
     5821 gagaaagctt gtttaagggg ccgcgtgatt acaacccgat atcgagcacc atttgtcatt
     5881 tgacgaatga atctgatggg cacacaacat cgttgtatgg tattggattt ggtcccttca
     5941 tcattacaaa caagcacttg tttagaagaa ataatggaac actgttggtc caatcactac
     6001 atggtgtatt caaggtcaag aacaccacga ctttgcaaca acacctcatt gatgggaggg
     6061 acatgataat tattcgcatg cctaaggatt tcccaccatt tcctcaaaag ctgaaattta
     6121 gagagccaca aagggaagag cgcatatgtc ttgtgacaac caacttccaa actaagagca
     6181 tgtctagcat ggtgtcagac actagttgca cattcccttc atctgatggc atattctgga
     6241 agcattggat tcaaaccaag gatgggcagt gtggcagtcc attagtatca actagagatg
     6301 ggttcattgt tggtatacac tcagcatcga atttcaccaa cacaaacaat tatttcacaa
     6361 gcgtgccgaa aaacttcatg gaattgttga caaatcagga ggcgcagcag tgggttagtg
     6421 gttggcgatt aaatgctgac tcagtattgt gggggggcca taaagttttc atggtgaaac
     6481 ctgaagagcc ttttcagcca gttaaggaag cgactcaact catgaatcgt cgtcgccgtc
     6541 gctaagtcca tcagcttgtc cagcagggtt gtccacgggc cgagcgaagc gagccagccg
     6601 gtggccgcgg aacaaaacta aagcgccaca agggcgcttt agtttgtttt cagtccagtt
     6661 acgctggagt ctgaggctcg tcctgaatga tatcaagctt
//
