LOCUS       sequence-8767-ex        6466 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 6466)
  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 6466)
  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: pTEK-4S-0X
FEATURES             Location/Qualifiers
     source          1..6466
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(465..484)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(1168..1184)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1169..1185)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             complement(4186..5001)
                     /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     4909..4928
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4993..5012
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5573..6161
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6062..6081
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6315..6332
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      6422..6449
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcggccgcgt ttaaacctgc aggcacgtgg catgcctcga ggatatcaga tctactagaa
       61 atattggatc gtcggcaccg tcacggccgt gggaggcggc acgatccgcg acgtgatgat
      121 cggccgcatc cccacggtgc tgcgcagtga gctctacgcc atcccggcgt tgatctgtgc
      181 gttcgcacgc acaggcccgg tgtgagaagg gtctctgacg agcgggagaa cccacccggg
      241 gtgggcgagt ttgtcctgcg tgtgctcggt cgagtaggct ctgggagtac ccgtgtgtac
      301 gaccagcacg gcatacatca tttcgacgcc gagagattcg ccgcccgaaa tgagcacgat
      361 ccgcggtacc atcaggagga atcacttcgc aatgtctaga ttagataaaa gtaaagtgat
      421 taacagcgca ttagagctgg gtaatgaggt cggaatcgaa ggtttaacaa cccgtaaact
      481 cgcccagaag cttggtgtag agcagcctac attgtattgg catgtaaaaa ataagcgggc
      541 cctactggat gcgctggcgg tggagatctt ggcgcgtcat catgattatt cactgcctgc
      601 ggcgggggaa tcctggcagt catttctgcg caataatgca atgagtttcc gccgcgcgct
      661 gctgcgttac cgtgacgggg caaaagtgca cctcggcacg cgtcctgatg aaaaacagta
      721 tgatacggtg gaaacccagt tacgctttat gacagaaaac ggcttttcac tgcgcgacgg
      781 gttatatgcg atttcagcgg tcagtcattt taccttaggt gccgtactgg agcagcagga
      841 gcatactgcc gccctgaccg accgccctgc agcaccggac gaaaacctgc cgccgctatt
      901 gcgggaagcg ctgcagatta tggacagtga tgatggtgag caggcctttc tgcatggcct
      961 ggagagcctg atccgggggt ttgaggtgca gcttacggca ctgttgcaaa tagtctgagg
     1021 ccatggtgac ctcatgactc gctagccagc tggggaccct agaggtcccc ttngaaaaaa
     1081 tttttaaaat aaaaaagggg acctctaggg tccccaatta aatagttaac actagtattt
     1141 aaatcaattg atgcatatta attaaatcga taccgtcgac ctcgaggggg ggcccgccgg
     1201 ggatcatcga gccgagaacg ttatcgaagt tggtcatgtg taatcccctc gtttgaactt
     1261 tggattaagc gtagatacac ccttggacaa gccagttgga ttcggagaca agcaaattca
     1321 gccttaaaaa gggcgaggcc tgcggtggtg gaacaccgca gggcctctaa ccgctcgacg
     1381 cgctgcacca accagcccgc gaacggctgg cagccagcgt aaggcgcggc tcatcgggcg
     1441 gcgttcgcca cgatgtcctg cacttcgagc caagcctcga acacctgctg gtgtgcacga
     1501 ctcacccggt tgttgacacc gcgcgcggcc gtgcgggctc ggtggggcgg ctgtgtcgcc
     1561 cttgccagcg tgagtagcgc gtacctcacc tcgcccaaca ggtcgcacac agccgattcg
     1621 tacgccataa agccaggtga gcccaccagc tccgtaagtt cgggcgctgt gtggctcgta
     1681 cccgcgcatt caggcggcag ggggtctaac gggtctaagg cggcgtgtac ggccgccaca
     1741 gcggctctca gcggcccgga aacgtcctcg aaacgacgca tgtgttcctc ctggttggta
     1801 caggtggttg ggggtgctcg gctgtcgctg gtgttccacc accagggctc gacgggagag
     1861 cgggggagtg tgcagttgtg gggtggcccc tcagcgaaat atctgacttg gagctcgtgt
     1921 cggaccatac accggtgatt aatcgtggtc tactaccaag cgtgagccac gtcgccgacg
     1981 aatttgagca gctctggctg ccgtactggc cgctggcaag cgacgatctg ctcgagggga
     2041 tctaccgcca aagccgcgcg tcggccctag gccgccggta catcgaggcg aacccaacag
     2101 cgctggcaaa cctgctggtc gtggacgtag accatccaga cgcagcgctc cgagcgctca
     2161 gcgcccgggg gtcccatccg ctgcccaacg cgatcgtggg caatcgcgcc aacggccacg
     2221 cacacgcagt gtgggcactc aacgcccctg ttccacgcac cgaatacgcg cggcgtaagc
     2281 cgctcgcata catggcggcg tgcgccgaag gccttcggcg cgccgtcgac ggcgaccgca
     2341 gttactcagg cctcatgacc aaaaaccccg gccacatcgc ctgggaaacg gaatggctcc
     2401 actcagatct ctacacactc agccacatcg aggccgagct cggcgcgaac atgccaccgc
     2461 cgcgctggcg tcagcagacc acgtacaaag cggctccgac gccgctaggg cggaattgcg
     2521 cactgttcga ttccgtcagg ttgtgggcct atcgtcccgc cctcatgcgg atctacctgc
     2581 cgacccggaa cgtggacgga ctcggccgcg cgatctatgc cgagtgccac gcgcgaaacg
     2641 ccgaattccc gtgcaacgac gtgtgtcccg gaccgctacc ggacagcgag gtccgcgcca
     2701 tcgccaacag catttggcgt tggatcacaa ccaagtcgcg catttgggcg gacgggatcg
     2761 tggtctacga ggccacactc agtgcgcgcc agtcggccat ctcgcggaag ggcgcagcag
     2821 cgcgcacggc ggcgagcaca gttgcgcggc gcgcaaagtc cgcgtcagcc atggaggcat
     2881 tgctatgagc gacggctaca gcgacggcta cagcgacggc tacaaccggc agccgactgt
     2941 ccgcaaaaag cggcgcgtga ccgccgccga aggcgctcga atcaccggac tatccgaacg
     3001 ccacgtcgtc cggctcgtgg cgcaggaacg cagcgagtgg ctcgccgagc aggctgcacg
     3061 ccgcgaacgc atccgcgcct atcacgacga cgagggccac tcttggccgc aaacggccaa
     3121 acatttcggg ctgcatctgg acaccgttaa gcgactcggc tatcgggcga ggaaagagcg
     3181 tgcggcagaa caggaagcgg ctcaaaaggc ccacaacgaa gccgacaatc caccgctgtt
     3241 ctaacgcaat tggggagcgg gtgtcgcggg ggttccgtgg ggggttccgt tgcaacgggt
     3301 cggacaggta aaagtcctgg tagacgctag ttttctggtt tgggccatgc ctgtctcgtt
     3361 gcgtgtttcg ttgcgcccgt tttgaatacc agccagacga gacggggttc tacgaatctt
     3421 ggtcgatacc aagccatttc cgctgaatat cggggagctc accgccagaa tcggtggttg
     3481 tggtgatgta cgtggcgaac tccgttgtag tgcctgtggt ggcatccgtg gccactctcg
     3541 ttgcacggtt cgttgtgccg ttacaggccc cgttgacagc tcaccgaacg tagttaaaac
     3601 atgctggtca aactaggttt accaacgata cgagtcagct catctagggc cagttctagg
     3661 cgttgttcgt tgcgcggttc gttgcgcatg tttcgtgtgg ttgctagatg gctccgcaac
     3721 cacacgcttc gaggttgagt gcttccagca cgggcgcgat ccagaagaac ttcgtcgtgc
     3781 gactgtcctc gttgatcctt gccgagctgg gatggaagct cggccgacca ccctggagga
     3841 gatgatcgag gatgccaggg cctttcacgc ccgccgctgc tgagcgtccg ccgccgggcc
     3901 cgcaccgccg tcggccggca gaaggtgttg ctgactcata ccaggcctga atcgccccat
     3961 catccagcca gaaagtgagg gagccacggt tgatgagagc tttgttgtag gtggaccagt
     4021 tggtgatttt gaacttttgc tttgccacgg aacggtctgc gttgtcggga agatgcgtga
     4081 tctgatcctt caactcagca aaagttcgat ttattcaaca aagccgccgt cccgtcaagt
     4141 cagcgtaatg ctctgccagt gttacaacca attaaccaat tctgattaga aaaactcatc
     4201 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
     4261 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     4321 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     4381 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     4441 tggcaaaaga ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     4501 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     4561 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     4621 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     4681 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     4741 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     4801 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatg
     4861 gggcttccca tacaagcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     4921 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctcg agcaagacgt
     4981 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
     5041 tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac
     5101 aacgtcgctt tgttgctagc tggcgggaac ggcaatgctc ctggcggcgg cggggctggt
     5161 gcccaggtct ccgcacagag cggcggtgct ggcgctcgcg gccaggcgtg gttcttcgcg
     5221 tactgacaag aaacccccct ctttaggact cagtgtcctt gggatcgaca cggcctgtcc
     5281 gcaccgcggt caggcgttga cgatgacggg ctggtcggcc acgtccgggg agtcaggcaa
     5341 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5401 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5461 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5521 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5581 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5641 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5701 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5761 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5821 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5881 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5941 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6001 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6061 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6121 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6181 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6241 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     6301 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     6361 cgcctctccc cgcgcgttgg ccgattcatt aatgcagcta gaactagttg attagctaag
     6421 cagaaggcca tcctgacgga tggccttttt gcgtttaata ctgttt
//
