LOCUS       sequence-450357-        6930 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express igr3 (Rv3542c-Rv3541c-Rv3540c) with K253E mutation in 
            Rv3542c in Mycobacteria .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6930)
  AUTHORS   Panagoda NT, Balazsi G, Sampson NS
  TITLE     Mycobacterium tuberculosis Mce3R TetR-like Repressor Forms an 
            Asymmetric Four-Helix Bundle and Binds a Nonpalindrome 
            Sequencedagger.
  JOURNAL   ACS Chem Biol. 2024 Dec 20;19(12):2580-2592. doi: 
            10.1021/acschembio.4c00687. Epub 2024 Nov 15.
  PUBMED    39545866
REFERENCE   2  (bases 1 to 6930)
  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: 
            "10.1021/acschembio.4c00687"; journalName: "ACS Chem Biol"; date: 
            "2024-12-20- 20"; volume: "19"; issue: "12"; pages: "2580-2592"
COMMENT     Sequence Label: pMV261-igr3K253E
FEATURES             Location/Qualifiers
     source          1..6930
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1389..1435
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1585..1604)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1596..2411
                     /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
                     KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1669..1688)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2742..3330
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3231..3250
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             5779..5808
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
ORIGIN
        1 ccgactgggc gggcccgtcg gcgctgatca agtcgattgg gctgcggctg ggggtgccgt
       61 ggtacgccta cgacacggtg acgttctccg gtgaggtgac cgcggtgaac gacggcttga
      121 tcacggtaaa ggtggtgggc cgcaataccc ttggcgacca cgtgaccgcg acggtggaac
      181 tttcgatgag ggattcgtag tgttatcggg tcaggcggcc atcgtcggta tcggcgccac
      241 cgacttttcg aagaactccg gtcgaagtga gctgcggctg gcggccgagg cggtgttgga
      301 tgcgttggcc gatgcgggcc tgagcccgac cgatgtcgac gggctgacca cgttcacgat
      361 ggacaccaac accgaaatcg ccgtggcgcg tgcggccggc atcggcgagc tgacgttctt
      421 ctccaagatc cactacggcg gtggcgccgc agcggcgacc gtgcagcacg ccgctatggc
      481 agtggccacc ggggtggctg acgtcgtggt ggcgtatcgg gcattcaacg aacgatccgg
      541 catgcggttc ggtcaggtgc aaactcgttt gaccgagaat gccgactcca ccggcgtgga
      601 caattcgttt tcgtatccgc acgggctctc cacgcccgcc gcgcaagtgg cgatgatcgc
      661 tcgccggtac atgcacctgt ctggtgcgac cagccgggac ttcggtgctg tctcggtggc
      721 cgaccgcaag catgccgcca acaaccccaa ggcgtacttc tacggcaagc cgataaccat
      781 tgaggaccac cagaattcga ggtggatcgc cgagccgctg cggctgctgg actgctgcca
      841 ggagaccgac ggcgcggtcg cgatcgtggt gacgtcagct gcgcgcgcac gggacctcaa
      901 gcagcgcccg gtggtcattg aggcggctgc gcagggctgc agtccagacc agtacacgat
      961 ggtcagctac taccggccgg aactcgacgg cctgcccgag atgggcctgg tgggccggca
     1021 gctatgggcg cagtcggggc tgacgccggc cgatgtccag accgcagtcc tctacgacca
     1081 cttcacgccg tttaccctga ttcagttgga ggagttggga ttctgcggca agggcgaagc
     1141 gaaagacttc atcgccgacg gcgcgatcga ggtgggcggg cggctgccca tcaacaccca
     1201 cggcggtcaa ctcggcgaag cctacatcca tggcatgaac ggcatcgcgg agggggtgcg
     1261 gcagctgcgc ggcacctcgg tgaacccggt ggcgggcgtc gagcatgtgc tcgtcaccgc
     1321 gggcaccggg gtgcctacgt ccgggctgat cctgggttaa aactagcgta cgatcgactg
     1381 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatgccat
     1441 catggccgcg gtgatcagct agccacctga cgtcgggggg gggggaaagc cacgttgtgt
     1501 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
     1561 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
     1621 cttgctcgag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
     1681 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagccccatg
     1741 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
     1801 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
     1861 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca gcattccagg
     1921 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
     1981 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc
     2041 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
     2101 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataatctt ttgccattct
     2161 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
     2221 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     2281 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     2341 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
     2401 agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt
     2461 gacgggacgg cggctttgtt gaataaatcg aacttttgct gagttgaagg atcagatcac
     2521 gcatcttccc gacaacgcag accgttccgt ggcaaagcaa aagttcaaaa tcaccaactg
     2581 gtccacctac aacaaagctc tcaccaaccg tggctccctc actttctggc tggatgatgg
     2641 ggcgattcag gcctggtatg agtcagcaac accttcttca cgaggcagac ctcactagtt
     2701 ccatgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2761 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2821 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2881 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2941 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3001 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3061 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3121 ctacagcgtg agctatgaga aagcgccacg cttcccgagg ggagaaaggc ggacaggtat
     3181 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3241 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3301 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3361 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3421 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3481 cgcaacgcgt gagcccacca gctccgtaag ttcgggcgct gtgtggctcg tacccgcgca
     3541 ttcaggcggc agggggtcta acgggtctaa ggcggcgtgt acggccgcca cagcggctct
     3601 cagcggcccg gaaacgtcct cgaaacgacg catgtgttcc tcctggttgg tacaggtggt
     3661 tgggggtgct cggctgtcgc tggtgttcca ccaccagggc tcgacgggag agcgggggag
     3721 tgtgcagttg tggggtggcc cctcagcgaa atatctgact tggagctcgt gtcggaccat
     3781 acaccggtga ttaatcgtgg tctactacca agcgtgagcc acgtcgccga cgaatttgag
     3841 cagctctggc tgccgtactg gccgctggca agcgacgatc tgctcgaggg gatctaccgc
     3901 caaagccgcg cgtcggccct aggccgccgg tacatcgagg cgaacccaac agcgctggca
     3961 aacctgctgg tcgtggacgt agaccatcca gacgcagcgc tccgagcgct cagcgcccgg
     4021 gggtcccatc cgctgcccaa cgcgatcgtg ggcaatcgcg ccaacggcca cgcacacgca
     4081 gtgtgggcac tcaacgcccc tgttccacgc accgaatacg cgcggcgtaa gccgctcgca
     4141 tacatggcgg cgtgcgccga aggccttcgg cgcgccgtcg atggcgaccg cagttactca
     4201 ggcctcatga ccaaaaaccc cggccacatc gcctgggaaa cggaatggct ccactcagat
     4261 ctctacacac tcagccacat cgaggccgag ctcggcgcga acatgccacc gccgcgctgg
     4321 cgtcagcaga ccacgtacaa agcggctccg acgccgctag ggcggaattg cgcactgttc
     4381 gattccgtca ggttgtgggc ctatcgtccc gccctcatgc ggatctacct gccgacccgg
     4441 aacgtggacg gactcggccg cgcgatctat gccgagtgcc acgcgcgaaa cgccgaattt
     4501 ccgtgcaacg acgtgtgtcc cggaccgcta ccggacagcg aggtccgcgc catcgccaac
     4561 agcatttggc gttggatcac aaccaagtcg cgcatttggg cggacgggat cgtggtctac
     4621 gaggccacac tcagtgcgcg ccagtcggcc atctcgcgga agggcgcagc agcgcgcacg
     4681 gcggcgagca cagttgcgcg gcgcgcaaag tccgcgtcag ccatggaggc attgctatga
     4741 gcgacggcta cagcgacggc tacaaccggc agccgactgt ccgcaaaaag cggcgcgtga
     4801 ccgccgccga aggcgctcga atcaccggac tatccgaacg ccacgtcgtc cggctcgtgg
     4861 cgcaggaacg cagcgagtgg ctcgccgagc aggctgcacg ccgcgaacgc atccgcgcct
     4921 atcacgacga cgagggccac tcttggccgc aaacggccaa acatttcggg ctgcatctgg
     4981 acaccgttaa gcgactcggc tatcgggcga ggaaagagcg tgcggcagaa caggaagcgg
     5041 ctcaaaaggc ccacaacgaa gccgacaatc caccgctgtt ctaacgcaat tggggagcgg
     5101 gtgtcgcggg ggttccgtgg ggggttccgt tgcaacgggt cggacaggta aaagtcctgg
     5161 tagacgctag ttttctggtt tgggccatgc ctgtctcgtt gcgtgtttcg ttgcgcccgt
     5221 tttgaatacc agccagacga gacggggttc tacgaatctt ggtcgatacc aagccatttc
     5281 cgctgaatat cggggagctc accgccagaa tcggtggttg tggtgatgta cgtggcgaac
     5341 tccgttgtag tgcctgtggt ggcatccgtg gcgcggccgc ggtaccagat ctttaaatct
     5401 agacggtgac cacaacgcgc ccgctttgat cggggacgtc tgcggccgac catttacggg
     5461 tcttgttgtc gttggcggtc atgggccgaa catactcacc cggatcggag ggccgaggac
     5521 aaggtcgaac gaggggcatg acccggtgcg gggcttcttg cactcggcat aggcgagtgc
     5581 taagaataac gttggcactc gcgaccggtg agtgctaggt cgggacggtg aggccaggcc
     5641 cgtcgtcgca gcgagtggca gcgaggacaa cttgagccgt ccgtcgcggg cactgcgccc
     5701 ggccagcgta agtagcgggg ttgccgtcac ccggtgaccc ccgtttcatc cccgatccgg
     5761 aggaatcact tcgcaatgga acaaaaactc atctcagaag aggatctggt gaccggggtg
     5821 agcgacattc aggaagcggt tgcccagatc aaggcggccg ggcccagcaa gccgcgcctg
     5881 gcccgtgacc cggtgaacca gccgatgatc aacaactggg tggaggccat cggcgaccgc
     5941 aaccccatct atgtggacga cgcggcggct cgcgccgccg gccaccccgg aatcgtcgcg
     6001 ccaccggcca tgattcaggt gtggaccatg atgggtctgg gcggagtgcg tccgaaggac
     6061 gatccgctgg gacccatcat caagctgttc gacgacgccg gctacatcgg tgtggttgcc
     6121 accaactgcg agcagaccta ccaccgctac ctattgcccg gggagcaagt cagcatcagc
     6181 gccgagctcg gtgacgtcgt cggacccaag caaaccgcgc tcggcgaggg ctggttcatc
     6241 aaccagcaca tcgtatggca agtcggggac gaggacgtcg cagagatgaa ttggcgcatc
     6301 ctcaaattca agcccgcggg ttctccgagc tcggtgcctg acgatttgga tcccgacgct
     6361 atgatgcgtc cctcgtcgtc gcgggacacc gcgtttttct gggatggagt caaggcccac
     6421 gaattgcgga tccagcggct tgccgatggc agcctgcggc acccaccggt gccggcggtg
     6481 tggcaagaca aatcggtgcc gatcaactac gtcgtctcca gcggtcgcgg cacggtgttc
     6541 agcttcgtgg tgcatcacgc gccggaggtt ccgggtcgca cggtgccctt cgtgatcgcg
     6601 ctgatggagc tcgaggaagg ggtacgcatg ctgggcgagc tgcggggggc cgatcctgct
     6661 cgtgttgcga tcggtatgcc ggtacgagcg acctatatcg atttccccga ctggagcctc
     6721 tacgcatggg agcctgacga atgacggttg ttggtgctgt gctgcctgag ctgaagctct
     6781 acggagaccc gacgttcatc gtctcaacgg cgctggctac cagggacttc caggatgtgc
     6841 atcatgatcg agacaaggcc gtcgcccagg ggtcgaaaga cattttcgtc aacatcctca
     6901 ccgacaccgg gctggtgcaa cgctacgtca
//
