LOCUS       sequence-9551-ex        5094 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 5094)
  AUTHORS   Klotzsche M, Ehrt S, Schnappinger D
  TITLE     Improved tetracycline repressors for gene silencing in mycobacteria.
  JOURNAL   Nucleic Acids Res. 2009 Apr;37(6):1778-88. doi: 10.1093/nar/gkp015. 
            Epub 2009 Jan 27.
  PUBMED    19174563
REFERENCE   2  (bases 1 to 5094)
  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: 
            "10.1093/nar/gkp015"; journalName: "Nucleic Acids Res"; date: 
            "2009-04"; volume: "37"; issue: "6"; pages: "1778-88"
COMMENT     Sequence Label: pTC-28S15-0X
FEATURES             Location/Qualifiers
     source          1..5094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1336..1924)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1416..1435)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2256..3071)
                     /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     2979..2998
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3063..3082
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(5014..5060)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gtcgacatcg atttaattaa tatgcatcaa ttgatttaaa tactagtgtt aactatttaa
       61 ttggggaccc tagaggtccc cttttttatt ttaaaaattt tttcnaaggg gacctctagg
      121 gtccccagct ggctagcgag tcatgaggtn accatggtca ttaggagccg ctctcgcact
      181 tcagctgctt ctccaagccg cagatgatca gctccaggcc gaacaagaag gccggctcgg
      241 cgccctggtg gtcgaacagc tcgatggcct ggcgcagcaa cggcggcatg ctgtcggtgg
      301 tcggggtctc gcgctcctcc ttggcggccg tgtgctcctg gtcctccaac acgcagccca
      361 gggtgaagtg gcccacggcc gacagggcgt acaacgcgtt ctccagcgag aagccgttct
      421 cggtcatgaa gcgcagctgg gtctccacgg tgtcgtactg cttctcgtcc gggcgggtgc
      481 ccaggtgcac cttggcgccg tcgcggtagc gcagcagcgc gcggcggaac gacatggcgt
      541 tgttgcgcag gaacgactgc cacgactcgc cggccgccgg cagcgagtag tcgtggtggc
      601 gcgccaggat ctccaccgcc agggcgtcca gcagggcccg cttgttcttg acgtgccagt
      661 acagggtcgg ctgctcgacg cccagcttct gggccagctt gcgggtggtc acgccctcga
      721 tgccgacctc gttgcccagg gccagggcgg agttgatgac cttggacttg tccaggcggg
      781 acattgcgaa gtgattcctc ctgatggtac cgcggatcgt gctcatttcg ggcggcgaat
      841 ctctcggcgt cgaaatgatg tatgccgtgc tggtcgtaca cacgggtact cccagagcct
      901 actcgaccga gcacacgcag gacaaactcg cccaccccgg gtgggttctc ccgctcgtca
      961 gagacccttc tcacaccggg cctgtgcgtg cgaacgcaca gatcaacgcc gggatggcgt
     1021 agagctcact gcgcagcacc gtggggatgc ggccgatcat cacgtcgcgg atcgtgccgc
     1081 ctcccacggc cgtgacggtg ccgacgatcc aatatttcta gtagatctga tatcctcgag
     1141 gcatgccacg tgcctgcagg tttaaacgcg gccgcacgcg ttgcgctcgg tcgttcggct
     1201 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1261 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1321 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1381 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1441 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1501 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
     1561 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1621 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1681 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1741 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     1801 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1861 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1921 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaact agtgaggtct
     1981 gcctcgtgaa gaaggtgttg ctgactcata ccaggcctga atcgccccat catccagcca
     2041 gaaagtgagg gagccacggt tgatgagagc tttgttgtag gtggaccagt tggtgatttt
     2101 gaacttttgc tttgccacgg aacggtctgc gttgtcggga agatgcgtga tctgatcctt
     2161 caactcagca aaagttcgat ttattcaaca aagccgccgt cccgtcaagt cagcgtaatg
     2221 ctctgccagt gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa
     2281 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     2341 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     2401 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     2461 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaaga
     2521 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     2581 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     2641 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     2701 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     2761 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     2821 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     2881 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatg gggcttccca
     2941 tacaagcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     3001 tataaatcag catccatgtt ggaatttaat cgcggcctcg agcaagacgt ttcccgttga
     3061 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
     3121 gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtcgctt
     3181 tgttgctagc tggcgggaac ggcaatgctc ctggcggcgg cggggctggt gcccaggtct
     3241 ccgcacagag cggcggtgct ggcgctcgcg gccaggcgtg gttcttcgcg tactgacaag
     3301 aaacccccct ctttaggact cagtgtcctt gggagggggg ctttttgcgt ttcaggaggt
     3361 cttggccagc ttggacatcg cctcagcgat agcctcgtcg cgggcctcag acgccatctg
     3421 gtacttcatc gccatcctag gagtcgtgtg accgagacgg gccatcagct ccttggtcgt
     3481 cgcacctgcc tgagcggcga acgtagcgcc gacagcgcgg aggtcgtgga tgcggagttc
     3541 cggccgaccg atcttggcgt agccacgctt cagcgacttg gtgaacgcgg acttcgacag
     3601 ccggttgccc tgcgtcgtgg tcaccaggaa tgcctcgggg cccttgttca tcttcgtacg
     3661 gtccttcatg tgcgctcgga tcatctccgc gacgtgaggc ggaaccgtca caggacgctt
     3721 cgaccggacg gtcttggcgt tgccaacgac gatcttgttc cccacgcggg aagcgccacg
     3781 gcgcacccgg agcttcatcg tcatgccgtc gtccacgatg tccttgcggc gaagctcgat
     3841 cagctctccg aaccggaggc tcgtccacgc caggatgtat gccgcgatcc ggtagtgctc
     3901 gaagatctca gcggcgacga tgtccagctc ctcaggcgtc agcgcctcta cgtcgcgctc
     3961 atcggctgcc ttctgctcga tccggcacgg gttctctgcg atcagcttgt cctcgaccgc
     4021 tgtgttcatc accgcccgga ggacgttgta ggcatgccgg cgggcagtcg ggtgcttcct
     4081 acccatcccg gcccaccacg cacgcaccag agctggcgtc atctctgtga ccgccacttc
     4141 acctagcacc gggtagatgc ggcgctccgc gtgcccgctg tacagatccc tggtgccgtc
     4201 tgcgaggtcg cgctccacga gccacttccg ggtgtactcc tccagcgtga tggcgctggc
     4261 ggctgccttc ttcgcccggt cctgtggagg ggtccaggtc tccatctcga tgagccgctt
     4321 ctcgcccgcg agccaggctt cggcgtccat cttgttgtcg taggtctgca gcgcgtagta
     4381 cctcacaccg tcctgcgggt tgacgtatga ggcttggatc ctcccgctgc gctgagtctt
     4441 cagcgatccc catccgcgac gtgccaacta ggtctcctct cgtcgtgaac aaggctaccg
     4501 ggttgcaact cctgtgcaac tctcaggctt caacgcgctt ctacgacctg caatttcttt
     4561 ccacttagag gatgcagccg agagggggta aaaacctatc ttgaccggcc catatatgtg
     4621 gtcggcagac acccattctt ccaaactagc tacgcgggtt cgattcccgt cgcccgctcc
     4681 gctggtcaga gggtgttttc gccctctggc catttttctt tccaggggtc tgcaactctt
     4741 gtgcgactct tctgacctgg gcatacgcgg ttgcaacgca tccctgatct ggctactttc
     4801 gatgctgaca aacgaataga gccccccgcc tgcgcgaaca gacgaggggc attcacacca
     4861 gattggagct ggtgcagtga agagaataga ccgggacaag gttgcaccgg gagttgcagc
     4921 ggtcggaacc ctcgccgtcg gcgggctggc gttcgccctg tcgttcacgg ctctagctga
     4981 tcaccgcggc catgatggcc ggacaaacaa cagataaaac gaaaggccca gtctttcgac
     5041 tgagcctttc gttttatttg atgcctggca gtcgatcgta cgctagttaa ctac
//
