LOCUS       sequence-8530-ex        5092 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5092)
  AUTHORS   Guo XV, Monteleone M, Klotzsche M, Kamionka A, Hillen W, Braunstein 
            M, Ehrt S, Schnappinger D
  TITLE     Silencing Mycobacterium smegmatis by using tetracycline repressors.
  JOURNAL   J Bacteriol. 2007 Jul . 189(13):4614-23.
  PUBMED    17483222
REFERENCE   2  (bases 1 to 5092)
  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: "J 
            Bacteriol. 2007 Jul . 189(13):4614-23."
COMMENT     Sequence Label: pMC1s
FEATURES             Location/Qualifiers
     source          1..5092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(70..693)
                     /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     601..620
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      complement(1055..1082)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1264..1852)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1344..1363)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2184..2999)
                     /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     2907..2926
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2991..3010
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(4942..4988)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggccgcaaaa taaaaaaggg gacctctagg gtccccaatt aattagctaa agccgctagc
       61 ccatggtcat taagacccac tttcacattt aagttgtttt tctaatccgc atatgatcaa
      121 ttcaaggccg aataagaagg ctggctctgc accttggtga tcaaataatt cgatagcttg
      181 tcgtaataat ggcggcatac tatcagtagt aggtgtttcc ctttcttctt tagcgacttg
      241 atgctcttga tcttccaata cgcaacctaa ggtaaaatgc cccacagcgc tgagtgcata
      301 taacgcgttc tctagtgaaa aaccttgttg gcataaaaag gctaattgat tttcgagagt
      361 ttcatactgt ttttctgtag gccgtgtacc taaatgtact tttgctccat cgcgatgact
      421 tagtaaagca catctaaaac ttttagcgtt attacgtaaa aaatcttgcc agctttcccc
      481 ttctaaaggg caaaagtgag tatggtgcct atctaacatc tcaatggcta aggcgtcgag
      541 cagggcccgc ttatttttta catgccaata caatgtaggc tgctctacac caagcttctg
      601 ggcgagttta cgggttgtta aaccttcgat tccgacctca ttaagcagct ctaatgcgct
      661 gttaatcact ttacttttat ctaatctaga cattgcgaag tgattcctcc tgatggtacc
      721 atgcggatcg tgctcatttc gggcggcgaa tctctcggcg tcgaaatgat gtatgccgtg
      781 ctggtcgtac acacgggtac tcccagagcc tactcgaccg agcacacgca ggacaaactc
      841 gcccaccccg ggtgggttct cccgctcgtc agagaccctt ctcacaccgg gcctgtgcgt
      901 gcgaacgcac agatcaacgc cgggatggcg tagagctcac tgcgcagcac cgtggggatg
      961 cggccgatca tcacgtcgcg gatcgtgccg cctcccacgg ccgtgacggt gccgacgatc
     1021 caatatttct agagtgcatg cagtattaaa cgcaaaaagg ccatccgtca ggatggcctt
     1081 ctgcttagct aatcagcggc cgcacgcgtt gcgctcggtc gttcggctgc ggcgagcggt
     1141 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1201 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1261 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1321 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1381 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1441 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1501 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1561 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1621 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1681 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     1741 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     1801 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     1861 tttgatcttt tctacggggt ctgacgctca gtggaactag tgaggtctgc ctcgtgaaga
     1921 aggtgttgct gactcatacc aggcctgaat cgccccatca tccagccaga aagtgaggga
     1981 gccacggttg atgagagctt tgttgtaggt ggaccagttg gtgattttga acttttgctt
     2041 tgccacggaa cggtctgcgt tgtcgggaag atgcgtgatc tgatccttca actcagcaaa
     2101 agttcgattt attcaacaaa gccgccgtcc cgtcaagtca gcgtaatgct ctgccagtgt
     2161 tacaaccaat taaccaattc tgattagaaa aactcatcga gcatcaaatg aaactgcaat
     2221 ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga
     2281 gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg
     2341 actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt
     2401 gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagatt atgcatttct
     2461 ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc
     2521 aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa
     2581 ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca
     2641 atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc
     2701 gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga
     2761 ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg
     2821 ctacctttgc catgtttcag aaacaactct ggcgcatggg gcttcccata caagcgatag
     2881 attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca
     2941 tccatgttgg aatttaatcg cggcctcgag caagacgttt cccgttgaat atggctcata
     3001 acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt
     3061 ttatcttgtg caatgtaaca tcagagattt tgagacacaa cgtcgctttg ttgctagctg
     3121 gcgggaacgg caatgctcct ggcggcggcg gggctggtgc ccaggtctcc gcacagagcg
     3181 gcggtgctgg cgctcgcggc caggcgtggt tcttcgcgta ctgacaagaa acccccctct
     3241 ttaggactca gtgtccttgg gaggggggct ttttgcgttt caggaggtct tggccagctt
     3301 ggacatcgcc tcagcgatag cctcgtcgcg ggcctcagac gccatctggt acttcatcgc
     3361 catcctagga gtcgtgtgac cgagacgggc catcagctcc ttggtcgtcg cacctgcctg
     3421 agcggcgaac gtagcgccga cagcgcggag gtcgtggatg cggagttccg gccgaccgat
     3481 cttggcgtag ccacgcttca gcgacttggt gaacgcggac ttcgacagcc ggttgccctg
     3541 cgtcgtggtc accaggaatg cctcggggcc cttgttcatc ttcgtacggt ccttcatgtg
     3601 cgctcggatc atctccgcga cgtgaggcgg aaccgtcaca ggacgcttcg accggacggt
     3661 cttggcgttg ccaacgacga tcttgttccc cacgcgggaa gcgccacggc gcacccggag
     3721 cttcatcgtc atgccgtcgt ccacgatgtc cttgcggcga agctcgatca gctctccgaa
     3781 ccggaggctc gtccacgcca ggatgtatgc cgcgatccgg tagtgctcga agatctcagc
     3841 ggcgacgatg tccagctcct caggcgtcag cgcctctacg tcgcgctcat cggctgcctt
     3901 ctgctcgatc cggcacgggt tctctgcgat cagcttgtcc tcgaccgctg tgttcatcac
     3961 cgcccggagg acgttgtagg catgccggcg ggcagtcggg tgcttcctac ccatcccggc
     4021 ccaccacgca cgcaccagag ctggcgtcat ctctgtgacc gccacttcac ctagcaccgg
     4081 gtagatgcgg cgctccgcgt gcccgctgta cagatccctg gtgccgtctg cgaggtcgcg
     4141 ctccacgagc cacttccggg tgtactcctc cagcgtgatg gcgctggcgg ctgccttctt
     4201 cgcccggtcc tgtggagggg tccaggtctc catctcgatg agccgcttct cgcccgcgag
     4261 ccaggcttcg gcgtccatct tgttgtcgta ggtctgcagc gcgtagtacc tcacaccgtc
     4321 ctgcgggttg acgtatgagg cttggatcct cccgctgcgc tgagtcttca gcgatcccca
     4381 tccgcgacgt gccaactagg tctcctctcg tcgtgaacaa ggctaccggg ttgcaactcc
     4441 tgtgcaactc tcaggcttca acgcgcttct acgacctgca atttctttcc acttagagga
     4501 tgcagccgag agggggtaaa aacctatctt gaccggccca tatatgtggt cggcagacac
     4561 ccattcttcc aaactagcta cgcgggttcg attcccgtcg cccgctccgc tggtcagagg
     4621 gtgttttcgc cctctggcca tttttctttc caggggtctg caactcttgt gcgactcttc
     4681 tgacctgggc atacgcggtt gcaacgcatc cctgatctgg ctactttcga tgctgacaaa
     4741 cgaatagagc cccccgcctg cgcgaacaga cgaggggcat tcacaccaga ttggagctgg
     4801 tgcagtgaag agaatagacc gggacaaggt tgcaccggga gttgcagcgg tcggaaccct
     4861 cgccgtcggc gggctggcgt tcgccctgtc gttcacggct ctagctgatc accgcggcca
     4921 tgatggccgg acaaacaaca gataaaacga aaggcccagt ctttcgactg agcctttcgt
     4981 tttatttgat gcctggcagt cgatcgtacg ctagttaact acgtcgacat cgataagctt
     5041 cgaattctgc agctggatcc atggatatct agatttaaag atctggtacc gc
//
