LOCUS       sequence-8768-ex        8026 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 8026)
  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 8026)
  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: pUV15tetORm
FEATURES             Location/Qualifiers
     source          1..8026
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      20..47
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             join(406..408,409..411,412..1125)
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(439..467)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1062..1082
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1179..1994
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             1179..1994
                     /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
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1252..1271)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     complement(2055..2071)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2056..2072)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      4896..4923
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             5108..5731
                     /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(5181..5200)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     rep_origin      7197..7785
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7686..7705
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7939..7956
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 actagttgat tagctaagca gaaggccatc ctgacggatg gcctttttgc gtttaatact
       61 gtttaaactc tagaaatatt ggatcgtcgg caccgtcacg gccgtgggag gcggcacgat
      121 ccgcgacgtg atgatcggcc gcatccccac ggtgctgcgc agtgagctct acgccatccc
      181 ggcgttgatc tgtgcgttcg cacgcacagg cccggtgtga gaagggtctc tgacgagcgg
      241 gagaactccc tatcagtgat agagtttgtc ctccctatca gtgatagata ggctctggga
      301 gtacccgtgt gtacgaccag cacggcatac atcatttcga cgccgagaga ttcgccgccc
      361 gaaatgagca cgatccgcat gcttaattaa gaaggagata tacatatggc tagcaaagga
      421 gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg
      481 cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcta catacggaaa gcttaccctt
      541 aaatttattt gcactactgg aaaactacct gttccatggc caacacttgt cactactttg
      601 acctatggtg ttcaatgctt ttcccgttat ccggatcata tgaaacggca tgactttttc
      661 aagagtgcca tgcccgaagg ttatgtacag gaacgcacta tatctttcaa agatgacggg
      721 aactacaaga cgcgtgctga agtcaagttt gaaggtgata cccttgttaa tcgtatcgag
      781 ttaaaaggta ttgattttaa agaagatgga aacattctcg gacacaaact cgagtacaac
      841 tataactcac acaatgtata catcacggca gacaaacaaa agaatggaat caaagctaac
      901 ttcaaaattc gccacaacat tgaagatggc tccgttcaac tagcagacca ttatcaacaa
      961 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtcgacacaa
     1021 tctgcccttt cgaaagatcc caacgaaaag cgtgaccaca tggtccttct tgagtttgta
     1081 actgctgctg ggattacaca tggcatggat gagctctaca aataatgaat tcgagcattt
     1141 aaatggctcc cccgggctgc agtaggagga ggggtgttat gagccatatt caacgggaaa
     1201 cgtcttgctc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     1261 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg
     1321 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     1381 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     1441 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc
     1501 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     1561 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     1621 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     1681 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataag cttttgccat
     1741 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     1801 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     1861 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     1921 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     1981 atgagttttt ctaatcagat atcaagcttt agctaattaa ttggggaccc tagaggtccc
     2041 cttttttatt ttatcgatac cgtcgacctc gagggggggc ccgccgggga tcatcgagcc
     2101 gagaacgtta tcgaagttgg tcatgtgtaa tcccctcgtt tgaactttgg attaagcgta
     2161 gatacaccct tggacaagcc agttggattc ggagacaagc aaattcagcc ttaaaaaggg
     2221 cgaggcctgc ggtggtggaa caccgcaggg cctctaaccg ctcgacgcgc tgcaccaacc
     2281 agcccgcgaa cggctggcag ccagcgtaag gcgcggctca tcgggcggcg ttcgccacga
     2341 tgtcctgcac ttcgagccaa gcctcgaaca cctgctggtg tgcacgactc acccggttgt
     2401 tgacaccgcg cgcggccgtg cgggctcggt ggggcggctg tgtcgccctt gccagcgtga
     2461 gtagcgcgta cctcacctcg cccaacaggt cgcacacagc cgattcgtac gccataaagc
     2521 caggtgagcc caccagctcc gtaagttcgg gcgctgtgtg gctcgtaccc gcgcattcag
     2581 gcggcagggg gtctaacggg tctaaggcgg cgtgtacggc cgccacagcg gctctcagcg
     2641 gcccggaaac gtcctcgaaa cgacgcatgt gttcctcctg gttggtacag gtggttgggg
     2701 gtgctcggct gtcgctggtg ttccaccacc agggctcgac gggagagcgg gggagtgtgc
     2761 agttgtgggg tggcccctca gcgaaatatc tgacttggag ctcgtgtcgg accatacacc
     2821 ggtgattaat cgtggtctac taccaagcgt gagccacgtc gccgacgaat ttgagcagct
     2881 ctggctgccg tactggccgc tggcaagcga cgatctgctc gaggggatct accgccaaag
     2941 ccgcgcgtcg gccctaggcc gccggtacat cgaggcgaac ccaacagcgc tggcaaacct
     3001 gctggtcgtg gacgtagacc atccagacgc agcgctccga gcgctcagcg cccgggggtc
     3061 ccatccgctg cccaacgcga tcgtgggcaa tcgcgccaac ggccacgcac acgcagtgtg
     3121 ggcactcaac gcccctgttc cacgcaccga atacgcgcgg cgtaagccgc tcgcatacat
     3181 ggcggcgtgc gccgaaggcc ttcggcgcgc cgtcgacggc gaccgcagtt actcaggcct
     3241 catgaccaaa aaccccggcc acatcgcctg ggaaacggaa tggctccact cagatctcta
     3301 cacactcagc cacatcgagg ccgagctcgg cgcgaacatg ccaccgccgc gctggcgtca
     3361 gcagaccacg tacaaagcgg ctccgacgcc gctagggcgg aattgcgcac tgttcgattc
     3421 cgtcaggttg tgggcctatc gtcccgccct catgcggatc tacctgccga cccggaacgt
     3481 ggacggactc ggccgcgcga tctatgccga gtgccacgcg cgaaacgccg aattcccgtg
     3541 caacgacgtg tgtcccggac cgctaccgga cagcgaggtc cgcgccatcg ccaacagcat
     3601 ttggcgttgg atcacaacca agtcgcgcat ttgggcggac gggatcgtgg tctacgaggc
     3661 cacactcagt gcgcgccagt cggccatctc gcggaagggc gcagcagcgc gcacggcggc
     3721 gagcacagtt gcgcggcgcg caaagtccgc gtcagccatg gaggcattgc tatgagcgac
     3781 ggctacagcg acggctacag cgacggctac aaccggcagc cgactgtccg caaaaagcgg
     3841 cgcgtgaccg ccgccgaagg cgctcgaatc accggactat ccgaacgcca cgtcgtccgg
     3901 ctcgtggcgc aggaacgcag cgagtggctc gccgagcagg ctgcacgccg cgaacgcatc
     3961 cgcgcctatc acgacgacga gggccactct tggccgcaaa cggccaaaca tttcgggctg
     4021 catctggaca ccgttaagcg actcggctat cgggcgagga aagagcgtgc ggcagaacag
     4081 gaagcggctc aaaaggccca caacgaagcc gacaatccac cgctgttcta acgcaattgg
     4141 ggagcgggtg tcgcgggggt tccgtggggg gttccgttgc aacgggtcgg acaggtaaaa
     4201 gtcctggtag acgctagttt tctggtttgg gccatgcctg tctcgttgcg tgtttcgttg
     4261 cgcccgtttt gaataccagc cagacgagac ggggttctac gaatcttggt cgataccaag
     4321 ccatttccgc tgaatatcgg ggagctcacc gccagaatcg gtggttgtgg tgatgtacgt
     4381 ggcgaactcc gttgtagtgc ctgtggtggc atccgtggcc actctcgttg cacggttcgt
     4441 tgtgccgtta caggccccgt tgacagctca ccgaacgtag ttaaaacatg ctggtcaaac
     4501 taggtttacc aacgatacga gtcagctcat ctagggccag ttctaggcgt tgttcgttgc
     4561 gcggttcgtt gcgcatgttt cgtgtggttg ctagatggct ccgcaaccac acgcttcgag
     4621 gttgagtgct tccagcacgg gcgcgatcca gaagaacttc gtcgtgcgac tgtcctcgtt
     4681 gatccttgcc gagctgggat ggaagctcgg ccgaccaccc tggaggagat gatcgaggat
     4741 gccagggcct ttcacgcccg ccgctgctga gcgtccgccg ccgggcccgc accgccgtcg
     4801 gccggcccgc tccgggctcg cagcagcggg cttcggcgcg ggcccggggc tcccgagcgc
     4861 gggcggggct ccgggcggcc gctgattagc taagcagaag gccatcctga cggatggcct
     4921 ttttgcgttt aatactgcat gcactctaga aatattggat cgtcgcaccg ggttaagcct
     4981 ggagtgcggt ggtgcctggt cggcattttc gcagtcgagg gctctcgtgt agcctgggcg
     5041 agttgccgac gcaggcgacc ctcntgccac ggatccgcat ggtaccatca ggaggaatca
     5101 cttcgcaatg tctagattag ataaaagtaa agtgattaac agcgcattag agctgcttaa
     5161 tgaggtcgga atcgaaggtt taacaacccg taaactcgcc cagaagcttg gtgtagagca
     5221 gcctacattg tattggcatg taaaaaataa gcgggccctg ctcgacgcct tagccattga
     5281 gatgttagat aggcaccata ctcacttttg ccctttagaa ggggaaagct ggcaagattt
     5341 tttacgtaat aacgctaaaa gttttagatg tgctttacta agtcatcgcg atggagcaaa
     5401 agtacattta ggtacacggc ctacagaaaa acagtatgaa actctcgaaa atcaattagc
     5461 ctttttatgc caacaaggtt tttcactaga gaacgcgtta tatgcactca gcgctgtggg
     5521 gcattttacc ttaggttgcg tattggaaga tcaagagcat caagtcgcta aagaagaaag
     5581 ggaaacacct actactgata gtatgccgcc attattacga caagctatcg aattatttga
     5641 tcaccaaggt gcagagccag ccttcttatt cggccttgaa ttgatcatat gcggattaga
     5701 aaaacaactt aaatgtgaaa gtgggtctta atgaccatgg gctagcggct ttagctaatt
     5761 aattggggac cctagaggtc ccctttttta ttttgcggcc gccgggggcc gggggcggcg
     5821 ccgggcggcc cggggcgtca ggcgccgggg gcggtgtccg gcggccccca gaggaactgc
     5881 gccagttcct ccggatcggt gaagccggag agatccagcg gggtctcctc gaacacctcg
     5941 aagtcgtgca ggaaggtgaa ggcgagcagt tcgcgggcga agtcctcggt ccgcttccac
     6001 tgcgccccgt cgagcagcgc ggccaggatc tcgcggtcgc cccggaaggc gttgagatgc
     6061 agttgcacca ggctgtagcg ggagtctccc gcatagacgt cggtgaagtc gacgatcccg
     6121 gtgacctcgg tcgcggccag gtccacgaag atgttggtcc cgtgcaggtc gccgtggacg
     6181 aaccggggtt cgcggccggc cagcagcgtg tccacgtccg gcagccagtc ctccaggcgg
     6241 tccagcagcc ggggcgagag gtagccccac ccgcggtggt cctcgacggt cgccgcgcgg
     6301 cgttcccgca gcagttccgg gaagacctcg gaatgggggg tgagcacggt gttcccggtc
     6361 agcggcaccc tgtgcagccg gccgagcacc cggccgagtt cgcgggccag ggcgagcagc
     6421 gcgttccggt cggtcgtgcc gtccatcgcg gaccgccagg tggtgccggt catccggctc
     6481 atcaccaggt agggccacgg ccaggctccg gtgccgggcc gcagctcgcc gcggccgagg
     6541 aggcggggca ccggcaccgg ggcgtccgcc aggaccgcgt acgcctccga ctccgacgcg
     6601 aggctctccg gaccgcacca gtgctcgccg aacagcttga tcaccgggtc gggctcgccg
     6661 accagtacgg ggttggtgct ctcgccgggc acccgcagca ccggcggcac cggcagcccg
     6721 agctcctcca gggctcggcg ggccagcggc tcccagaatt cctggtcgtt ccgcaggctc
     6781 gcgtaggaat catccgaatc aatacggtcg agaagtaaca gggattcttg tgtcacagcg
     6841 gacctctatt cacagggtac gggccggctt aattccgcac ggccggtcgc gacacggcct
     6901 gtccgcaccg cggtcaggcg ttgacgatga cgggctggtc ggccacgtcc ggggagtcag
     6961 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     7021 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     7081 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     7141 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     7201 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     7261 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7321 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     7381 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7441 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     7501 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7561 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7621 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7681 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7741 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7801 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     7861 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     7921 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     7981 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctaga
//
