LOCUS       sequence-9552-ex        3955 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 3955)
  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 3955)
  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-mcs
FEATURES             Location/Qualifiers
     source          1..3955
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(37..83)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(314..902)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(394..413)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1234..2049)
                     /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     1957..1976
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2041..2060
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tgatcaccgc ggccatgatg gccggacaaa caacagataa aacgaaaggc ccagtctttc
       61 gactgagcct ttcgttttat ttgatgcctg gcagtcgatc gtacgctagt taactacgtc
      121 gacatcgatt taattaagtt taaacgcggc cgcacgcgtt gcgctcggtc gttcggctgc
      181 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
      241 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      301 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      361 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      421 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      481 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
      541 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
      601 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      661 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
      721 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
      781 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
      841 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
      901 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaactag tgaggtctgc
      961 ctcgtgaaga aggtgttgct gactcatacc aggcctgaat cgccccatca tccagccaga
     1021 aagtgaggga gccacggttg atgagagctt tgttgtaggt ggaccagttg gtgattttga
     1081 acttttgctt tgccacggaa cggtctgcgt tgtcgggaag atgcgtgatc tgatccttca
     1141 actcagcaaa agttcgattt attcaacaaa gccgccgtcc cgtcaagtca gcgtaatgct
     1201 ctgccagtgt tacaaccaat taaccaattc tgattagaaa aactcatcga gcatcaaatg
     1261 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     1321 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     1381 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     1441 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagatt
     1501 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     1561 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
     1621 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     1681 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     1741 cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     1801 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     1861 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatggg gcttcccata
     1921 caagcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     1981 taaatcagca tccatgttgg aatttaatcg cggcctcgag caagacgttt cccgttgaat
     2041 atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga
     2101 tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacaa cgtcgctttg
     2161 ttgctagctg gcgggaacgg caatgctcct ggcggcggcg gggctggtgc ccaggtctcc
     2221 gcacagagcg gcggtgctgg cgctcgcggc caggcgtggt tcttcgcgta ctgacaagaa
     2281 acccccctct ttaggactca gtgtccttgg gaggggggct ttttgcgttt caggaggtct
     2341 tggccagctt ggacatcgcc tcagcgatag cctcgtcgcg ggcctcagac gccatctggt
     2401 acttcatcgc catcctagga gtcgtgtgac cgagacgggc catcagctcc ttggtcgtcg
     2461 cacctgcctg agcggcgaac gtagcgccga cagcgcggag gtcgtggatg cggagttccg
     2521 gccgaccgat cttggcgtag ccacgcttca gcgacttggt gaacgcggac ttcgacagcc
     2581 ggttgccctg cgtcgtggtc accaggaatg cctcggggcc cttgttcatc ttcgtacggt
     2641 ccttcatgtg cgctcggatc atctccgcga cgtgaggcgg aaccgtcaca ggacgcttcg
     2701 accggacggt cttggcgttg ccaacgacga tcttgttccc cacgcgggaa gcgccacggc
     2761 gcacccggag cttcatcgtc atgccgtcgt ccacgatgtc cttgcggcga agctcgatca
     2821 gctctccgaa ccggaggctc gtccacgcca ggatgtatgc cgcgatccgg tagtgctcga
     2881 agatctcagc ggcgacgatg tccagctcct caggcgtcag cgcctctacg tcgcgctcat
     2941 cggctgcctt ctgctcgatc cggcacgggt tctctgcgat cagcttgtcc tcgaccgctg
     3001 tgttcatcac cgcccggagg acgttgtagg catgccggcg ggcagtcggg tgcttcctac
     3061 ccatcccggc ccaccacgca cgcaccagag ctggcgtcat ctctgtgacc gccacttcac
     3121 ctagcaccgg gtagatgcgg cgctccgcgt gcccgctgta cagatccctg gtgccgtctg
     3181 cgaggtcgcg ctccacgagc cacttccggg tgtactcctc cagcgtgatg gcgctggcgg
     3241 ctgccttctt cgcccggtcc tgtggagggg tccaggtctc catctcgatg agccgcttct
     3301 cgcccgcgag ccaggcttcg gcgtccatct tgttgtcgta ggtctgcagc gcgtagtacc
     3361 tcacaccgtc ctgcgggttg acgtatgagg cttggatcct cccgctgcgc tgagtcttca
     3421 gcgatcccca tccgcgacgt gccaactagg tctcctctcg tcgtgaacaa ggctaccggg
     3481 ttgcaactcc tgtgcaactc tcaggcttca acgcgcttct acgacctgca atttctttcc
     3541 acttagagga tgcagccgag agggggtaaa aacctatctt gaccggccca tatatgtggt
     3601 cggcagacac ccattcttcc aaactagcta cgcgggttcg attcccgtcg cccgctccgc
     3661 tggtcagagg gtgttttcgc cctctggcca tttttctttc caggggtctg caactcttgt
     3721 gcgactcttc tgacctgggc atacgcggtt gcaacgcatc cctgatctgg ctactttcga
     3781 tgctgacaaa cgaatagagc cccccgcctg cgcgaacaga cgaggggcat tcacaccaga
     3841 ttggagctgg tgcagtgaag agaatagacc gggacaaggt tgcaccggga gttgcagcgg
     3901 tcggaaccct cgccgtcggc gggctggcgt tcgccctgtc gttcacggct ctagc
//
