LOCUS       sequence-266168-        7036 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To generate an integrative plasmid carrying the TetR/Pip-OFF system,
            but not the integrase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7036)
  AUTHORS   Boldrin F, Anoosheh S, Serafini A, Cioetto Mazzabo L, Palu G, 
            Provvedi R, Manganelli R
  TITLE     Improving the stability of the TetR/Pip-OFF mycobacterial 
            repressible promoter system.
  JOURNAL   Sci Rep. 2019 Apr 8;9(1):5783. doi: 10.1038/s41598-019-42319-2.
  PUBMED    30962489
REFERENCE   2  (bases 1 to 7036)
  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: "2019-04-8"; volume: "9(1):5783. doi"; pages: " 
            10.1038/s41598-019-42319-2"
COMMENT     Sequence Label: pANTO5
FEATURES             Location/Qualifiers
     source          1..7036
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..624
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /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="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(74..93)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(876..895)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(960..979)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             2006..2797
                     /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      3975..4563
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4464..4483
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(5437..5483)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(5845..5856)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 atgtctagat tagataaaag taaagtgatt aacagcgcat tagagctgct taatgaggtc
       61 ggaatcgaag gtttaacaac ccgtaaactc gcccagaagc ttggtgtaga gcagcctaca
      121 ttgtattggc atgtaaaaaa taagcgggcc ctgctcgacg ccttagccat tgagatgtta
      181 gataggcacc atactcactt ttgcccttta gaaggggaaa gctggcaaga ttttttacgt
      241 aataacgcta aaagttttag atgtgcttta ctaagtcatc gcgatggagc aaaagtacat
      301 ttaggtacac ggcctacaga aaaacagtat gaaactctcg aaaatcaatt agccttttta
      361 tgccaacaag gtttttcact agagaacgcg ttatatgcac tcagcgctgt ggggcatttt
      421 accttaggtt gcgtattgga agatcaagag catcaagtcg ctaaagaaga aagggaaaca
      481 cctactactg atagtatgcc gccattatta cgacaagcta tcgaattatt tgatcaccaa
      541 ggtgcagagc cagccttctt attcggcctt gaattgatca tatgcggatt agaaaaacaa
      601 cttaaatgtg aaagtgggtc ttaatgacca tgggctagcg ctttagctaa ttaattgggg
      661 accctagagg tccccttttt tattttgcgg ccctagtgga tccgagctcg gtaccaagct
      721 tgacgcggat ccgcatgcgg taccaagctc tagccacctg acgtcggggg ggggggaaag
      781 ccacgttgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
      841 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
      901 acgggaaacg tcttgctcga ggccgcgatt aaattccaac atggatgctg atttatatgg
      961 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     1021 gaagccccat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     1081 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     1141 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccaaaataaa
     1201 aaaggggacc tctagggtcc ccaattaatt agtaatataa tctattaaag gtcattcaaa
     1261 aggtcatcca ccggatcaat tcccctgctc gcgcaggctg ggtgccaagc tctcgggtaa
     1321 catcaaggcc cgatccttgg agcccttgcc ctcccgcacg atgatcgtgc cgtgatcgaa
     1381 atccagatcc ttgacccgca gttgcaaacc ctcactgatc cgcatgcccg ttccatacag
     1441 aagctgggcg aacaaacgat gctcgccttc cagaaaaccg aggatgcgaa ccacttcatc
     1501 cggggtcagc accaccggca agcgccgcga cggccgaggt cttccgatct cctgaagcca
     1561 gggcagatcc gtgcacagca ccttgccgta gaagaacagc aaggccgcca atgcctgacg
     1621 atgcgtggag accgaaacct tgcgctcgtt cgccagccag gacagaaatg cctcgacttc
     1681 gctgctgccc aaggttgccg ggtgacgcac accgtggaaa cggatgaagg cacgaaccca
     1741 gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa
     1801 ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc
     1861 ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg
     1921 ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag
     1981 tcgccctaaa acaaagttaa acatcatgag ggaagcggtg atcgccgaag tatcgactca
     2041 actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca
     2101 tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt
     2161 tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga
     2221 aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt
     2281 gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg
     2341 gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc
     2401 tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga
     2461 actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct
     2521 atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg
     2581 catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc
     2641 aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct
     2701 tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta
     2761 cgtgaaaggc gagatcacca aggtagtcgg caaataatgt ctaacaattc gttcaagccg
     2821 acgccgcttc gcggcgcggc ttaactcaag cgttagatgc actaagcaca taattgctca
     2881 cagccaaact atcaggtcaa gtctgctttt attattttta agcgtgcata ataagcccta
     2941 cacaaattgg gagatatatc atgaaaggct ggctttttct tgttatcgca atagttggcg
     3001 aagtaatcgc aacatccgca ttaaaatcta gcgagggctt tactaagctg atccggtgga
     3061 tgaccttttg aatgaccttt aatagattat attactaatt aattggggac cctagaggtc
     3121 ccctttttta ttttgggaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     3181 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     3241 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     3301 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     3361 tctggaaaga aatgcataat cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     3421 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     3481 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     3541 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     3601 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
     3661 ggttgtaaca ctggcagagc attacgctga cttgacggga cggcggcttt gttgaataaa
     3721 tcgaactttt gctgagttga aggatcagat cacgcatctt cccgacaacg cagaccgttc
     3781 cgtggcaaag caaaagttca aaatcaccaa ctggtccacc tacaacaaag ctctcaccaa
     3841 ccgtggctcc ctcactttct ggctggatga tggggcgatt caggcctggt atgagtcagc
     3901 aacaccttct tcacgaggca gacctcacta gttccatgag cgtcagaccc cgtagaaaag
     3961 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4021 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4081 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     4141 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4201 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4261 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4321 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     4381 acgcttcccg aggggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4441 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4501 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4561 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4621 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     4681 gctgataccg ctcgccgcag ccgaacgacc gagcgcaacg cgtgcggccg cggtaccaga
     4741 tctttgtgga tgacctttta tagattatat tactaattaa ttggggaccc tagaggtccc
     4801 cttttttatt ttaaaaattt tttcacaaaa cggtttacaa gcataaagct agtgatcctc
     4861 ccgctgcgct gagtcttcag cgatccccat ccgcgacgtg ccaactaggt ctcctctcgt
     4921 cgtgaacaag gctaccgggt tgcaactcct gtgcaactct caggcttcaa cgcgcttcta
     4981 cgacctgcaa tttctttcca cttagaggat gcagccgaga gggggtaaaa acctatcttg
     5041 accggcccat atatgtggtc ggcagacacc cattcttcca aactagctac gcgggttcga
     5101 ttcccgtcgc ccgctccgct ggtcagaggg tgttttcgcc ctctggccat ttttctttcc
     5161 aggggtctgc aactcttgtg cgactcttct gacctgggca tacgcggttg caacgcatcc
     5221 ctgatctggc tactttcgat gctgacaaac gaatagagcc ccccgcctgc gcgaacagac
     5281 gaggggcatt cacaccagat tggagctggt gcagtgaaga gaatagaccg ggacaaggtt
     5341 gcaccgggag ttgcagcggt cggaaccctc gccgtcggcg ggctggcgtt cgccctgtcg
     5401 ttcacggctc tagctgatca ccgcggccat gatggcataa aacgaaaggc ccagtctttc
     5461 gactgagcct ttcgttttat ttgatgcctg gcagtcgatc gtacgctagt taactacgtc
     5521 gacatcgata agcttcgaat tctgcagctg gatcactgtg ctcgagcggc cgccagtgtg
     5581 atggatatct gcagaattcg cccttaagct tttaattaat caggcctgtt cgaccatcgc
     5641 gtcgatgccc gcgacgagca ggtcgagggc gaactcgaag tcccggtcca gcatctccgc
     5701 cacggtgtcg ccgccccggg ccgccatgat gtcctgcgcg tcctcgatga cgcccgcggt
     5761 gtccggcacc tcggtcaccg cggtcatcga gtcctggaag tactcctccg gactcagccc
     5821 ggtgtccgcc acccgggcga ggaagcggcc ctcgatggtg ccgtagccgt agacgaactg
     5881 gaagacggcc gagatggcgc cggtcaggcg gtgcgcgggc agcccgctgc ggcgcacgac
     5941 gttctgcacc gcgcgggaga aggccagcga gtgcgggccg atgttgaggt aggtgccgac
     6001 cagccgggac gaccaggggt ggcgcaccag cagcgcccgg ttctcccggg ccagggcccg
     6061 cagttcctcg cgccagtcga gcccggcgtc cgggtccggg tggcgcagct cgccgaagac
     6121 ggcgtccagg gcgagctcga gcaactggtc cttggtgtcg acgtaccagt acacggacat
     6181 cgcggtgacg ttcagctcgg cggccaggcg gcgcatcgag aaccccgtca ggccctccgt
     6241 gtccagcagc cggacggtga ccccggtgat ccggtcccgg tcgagcccgg acggctgccc
     6301 cccacggcga ccgccgcgcc gcccctcccc cgacagccac acgctgtccc gcggcccctc
     6361 ccgccctgcc ttcgccatgc gcacctctcc tcgactcgcg gccgccacac tagacaatat
     6421 gtctatcact gatagggatc agttgatcta tcactgatag ggaggcgtct gctaggaccc
     6481 gatcgccccg gggccgctgg atcgtgggat ggcgggtgga tcagccttcg tatgttccga
     6541 tgagccgggc ctgcatggtg gaaggccatc cgtcaggatg ggatatcaag cttaagggcg
     6601 aattccagca cactggcggc cgttactagg gccgctgatt agctaagcag aaggccatcc
     6661 tgacggatgc ctttttcgct ttaatactgc atgcactcta gaaatattgg atcgtcggca
     6721 ccgtcacggc cgtgggaggc ggcacgatcc gcgacgtgat gatcggccgc atccccacgg
     6781 tgctgcgcag tgagctctac gccatcccgg cgttgatctg tgcgttcgca cgcacaggcc
     6841 cggtgtgaga agggtctctg acgagcggga gaacccaccc ggggtgggcg agtttgtcct
     6901 gcgtgtgctc ggtcgagtag gctctgggag tacccgtgtg tacgaccagc acggcataca
     6961 tcatttcgac gccgagagat tcgccgcccg aaatgagcac gatccgcatg gtaccatcag
     7021 gaggaatcac ttcgca
//
