LOCUS       sequence-9550-ex        7945 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 7945)
  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 7945)
  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-0X1L
FEATURES             Location/Qualifiers
     source          1..7945
                     /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"
     regulatory      269..278
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             663..3710
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
                     LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP
                     FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
                     DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
                     FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
                     TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
                     LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
                     YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
                     NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
                     LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
                     WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
                     FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
                     PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
                     SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
                     ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
                     ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR
                     CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
                     SWSPSVSAEFQLSAGRYHYQLVWCQK"
     primer_bind     complement(665..687)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    3974..4108
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4011..4030)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4065..4084
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4304..4892)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4384..4403)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5224..6039)
                     /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     5947..5966
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6031..6050
                     /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 taattaatat gcatcaattg atttaaatac tagtgttaac tatttaattg
      181 gggaccctag aggtcccctt ttttatttta aaaatttttt cnaaggggac ctctagggtc
      241 cccagctggc tagcgagtca tgaggtnacc atggtggcca tctagacatc atcggtacca
      301 ggcctagatc tactagaaat attggatcgt cggcaccgtc acggccgtgg gaggcggcac
      361 gatccgcgac gtgatgatcg gccgcatccc cacggtgctg cgcagtgagc tctacgccat
      421 cccggcgttg atctgtgcgt tcgcacgcac aggcccggtg tgagaagggt ctctgacgag
      481 cgggagaact ccctatcagt gatagagttt gtcctcccta tcagtgatag ataggctctg
      541 ggagtacccg tgtgtacgac cagcacggca tacatcattt cgacgccgag agattcgccg
      601 cccgaaatga gcacgatccg ctcgaggcat gcgaaggaga tatacatatg gagatcgatc
      661 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
      721 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
      781 cccaacagtt gcgcagcctg aatggcgaat ggcgctttgc ctggtttccg gcaccagaag
      841 cggtgccgga aagctggctg gagtgcgatc ttcctgaggc cgatactgtc gtcgtcccct
      901 caaactggca gatgcacggt tacgatgcgc ccatctacac caacgtaacc tatcccatta
      961 cggtcaatcc gccgtttgtt cccacggaga atccgacggg ttgttactcg ctcacattta
     1021 atgttgatga aagctggcta caggaaggcc agacgcgaat tatttttgat ggcgttaact
     1081 cggcgtttca tctgtggtgc aacgggcgct gggtcggtta cggccaggac agtcgtttgc
     1141 cgtctgaatt tgacctgagc gcatttttac gcgccggaga aaaccgcctc gcggtgatgg
     1201 tgctgcgttg gagtgacggc agttatctgg aagatcagga tatgtggcgg atgagcggca
     1261 ttttccgtga cgtctcgttg ctgcataaac cgactacaca aatcagcgat ttccatgttg
     1321 ccactcgctt taatgatgat ttcagccgcg ctgtactgga ggctgaagtt cagatgtgcg
     1381 gcgagttgcg tgactaccta cgggtaacag tttctttatg gcagggtgaa acgcaggtcg
     1441 ccagcggcac cgcgcctttc ggcggtgaaa ttatcgatga gcgtggtggt tatgccgatc
     1501 gcgtcacact acgtctgaac gtcgaaaacc cgaaactgtg gagcgccgaa atcccgaatc
     1561 tctatcgtgc ggtggttgaa ctgcacaccg ccgacggcac gctgattgaa gcagaagcct
     1621 gcgatgtcgg tttccgcgag gtgcggattg aaaatggtct gctgctgctg aacggcaagc
     1681 cgttgctgat tcgaggcgtt aaccgtcacg agcatcatcc tctgcatggt caggtcatgg
     1741 atgagcagac gatggtgcag gatatcctgc tgatgaagca gaacaacttt aacgccgtgc
     1801 gctgttcgca ttatccgaac catccgctgt ggtacacgct gtgcgaccgc tacggcctgt
     1861 atgtggtgga tgaagccaat attgaaaccc acggcatggt gccaatgaat cgtctgaccg
     1921 atgatccgcg ctggctaccg gcgatgagcg aacgcgtaac gcgaatggtg cagcgcgatc
     1981 gtaatcaccc gagtgtgatc atctggtcgc tggggaatga atcaggccac ggcgctaatc
     2041 acgacgcgct gtatcgctgg atcaaatctg tcgatccttc ccgcccggtg cagtatgaag
     2101 gcggcggagc cgacaccacg gccaccgata ttatttgccc gatgtacgcg cgcgtggatg
     2161 aagaccagcc cttcccggct gtgccgaaat ggtccatcaa aaaatggctt tcgctacctg
     2221 gagagacgcg cccgctgatc ctttgcgaat acgcccacgc gatgggtaac agtcttggcg
     2281 gtttcgctaa atactggcag gcgtttcgtc agtatccccg tttacagggc ggcttcgtct
     2341 gggactgggt ggatcagtcg ctgattaaat atgatgaaaa cggcaacccg tggtcggctt
     2401 acggcggtga ttttggcgat acgccgaacg atcgccagtt ctgtatgaac ggtctggtct
     2461 ttgccgaccg cacgccgcat ccagcgctga cggaagcaaa acaccagcag cagtttttcc
     2521 agttccgttt atccgggcaa accatcgaag tgaccagcga atacctgttc cgtcatagcg
     2581 ataacgagct cctgcactgg atggtggcgc tggatggtaa gccgctggca agcggtgaag
     2641 tgcctctgga tgtcgctcca caaggtaaac agttgattga actgcctgaa ctaccgcagc
     2701 cggagagcgc cgggcaactc tggctcacag tacgcgtagt gcaaccgaac gcgaccgcat
     2761 ggtcagaagc cgggcacatc agcgcctggc agcagtggcg tctggcggaa aacctcagtg
     2821 tgacgctccc cgccgcgtcc cacgccatcc cgcatctgac caccagcgaa atggattttt
     2881 gcatcgagct gggtaataag cgttggcaat ttaaccgcca gtcaggcttt ctttcacaga
     2941 tgtggattgg cgataaaaaa caactgctga cgccgctgcg cgatcagttc acccgtgcac
     3001 cgctggataa cgacattggc gtaagtgaag cgacccgcat tgaccctaac gcctgggtcg
     3061 aacgctggaa ggcggcgggc cattaccagg ccgaagcagc gttgttgcag tgcacggcag
     3121 atacacttgc tgatgcggtg ctgattacga ccgctcacgc gtggcagcat caggggaaaa
     3181 ccttatttat cagccggaaa acctaccgga ttgatggtag tggtcaaatg gcgattaccg
     3241 ttgatgttga agtggcgagc gatacaccgc atccggcgcg gattggcctg aactgccagc
     3301 tggcgcaggt agcagagcgg gtaaactggc tcggattagg gccgcaagaa aactatcccg
     3361 accgccttac tgccgcctgt tttgaccgct gggatctgcc attgtcagac atgtataccc
     3421 cgtacgtctt cccgagcgaa aacggtctgc gctgcgggac gcgcgaattg aattatggcc
     3481 cacaccagtg gcgcggcgac ttccagttca acatcagccg ctacagtcaa cagcaactga
     3541 tggaaaccag ccatcgccat ctgctgcacg cggaagaagg cacatggctg aatatcgacg
     3601 gtttccatat ggggattggt ggcgacgact cctggagccc gtcagtatcg gcggaattcc
     3661 agctgagcgc cggtcgctac cattaccagt tggtctggtg tcaaaaataa taataaccgg
     3721 gcaggccatg tctgcccgta tttcgcgtaa ggaaatccat tatgtactat ttaaaaaaca
     3781 caaacttttg gatgttcggt ttattctttt tcttttactt ttttatcatg ggagcctact
     3841 tcccgttttt cccgatttgg ctacatgaca tcaaccatat cagcaaaagt gatacgggta
     3901 ttatttttgc cgctatttct ctgttctcgc tattattcca accgctgttt ggtctgcttt
     3961 ctgacaaact cggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4021 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4081 catcaatgta tcttatcatg tctggatcct ctagagtcga cctgcaggtt taaacgcggc
     4141 cgcacgcgtt gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4201 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4261 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4321 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4381 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4441 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
     4501 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4561 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4621 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4681 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4741 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4801 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4861 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4921 ctgacgctca gtggaactag tgaggtctgc ctcgtgaaga aggtgttgct gactcatacc
     4981 aggcctgaat cgccccatca tccagccaga aagtgaggga gccacggttg atgagagctt
     5041 tgttgtaggt ggaccagttg gtgattttga acttttgctt tgccacggaa cggtctgcgt
     5101 tgtcgggaag atgcgtgatc tgatccttca actcagcaaa agttcgattt attcaacaaa
     5161 gccgccgtcc cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat taaccaattc
     5221 tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
     5281 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     5341 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     5401 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     5461 gactgaatcc ggtgagaatg gcaaaagatt atgcatttct ttccagactt gttcaacagg
     5521 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     5581 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
     5641 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
     5701 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
     5761 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     5821 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     5881 aaacaactct ggcgcatggg gcttcccata caagcgatag attgtcgcac ctgattgccc
     5941 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     6001 cggcctcgag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
     6061 tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca
     6121 tcagagattt tgagacacaa cgtcgctttg ttgctagctg gcgggaacgg caatgctcct
     6181 ggcggcggcg gggctggtgc ccaggtctcc gcacagagcg gcggtgctgg cgctcgcggc
     6241 caggcgtggt tcttcgcgta ctgacaagaa acccccctct ttaggactca gtgtccttgg
     6301 gaggggggct ttttgcgttt caggaggtct tggccagctt ggacatcgcc tcagcgatag
     6361 cctcgtcgcg ggcctcagac gccatctggt acttcatcgc catcctagga gtcgtgtgac
     6421 cgagacgggc catcagctcc ttggtcgtcg cacctgcctg agcggcgaac gtagcgccga
     6481 cagcgcggag gtcgtggatg cggagttccg gccgaccgat cttggcgtag ccacgcttca
     6541 gcgacttggt gaacgcggac ttcgacagcc ggttgccctg cgtcgtggtc accaggaatg
     6601 cctcggggcc cttgttcatc ttcgtacggt ccttcatgtg cgctcggatc atctccgcga
     6661 cgtgaggcgg aaccgtcaca ggacgcttcg accggacggt cttggcgttg ccaacgacga
     6721 tcttgttccc cacgcgggaa gcgccacggc gcacccggag cttcatcgtc atgccgtcgt
     6781 ccacgatgtc cttgcggcga agctcgatca gctctccgaa ccggaggctc gtccacgcca
     6841 ggatgtatgc cgcgatccgg tagtgctcga agatctcagc ggcgacgatg tccagctcct
     6901 caggcgtcag cgcctctacg tcgcgctcat cggctgcctt ctgctcgatc cggcacgggt
     6961 tctctgcgat cagcttgtcc tcgaccgctg tgttcatcac cgcccggagg acgttgtagg
     7021 catgccggcg ggcagtcggg tgcttcctac ccatcccggc ccaccacgca cgcaccagag
     7081 ctggcgtcat ctctgtgacc gccacttcac ctagcaccgg gtagatgcgg cgctccgcgt
     7141 gcccgctgta cagatccctg gtgccgtctg cgaggtcgcg ctccacgagc cacttccggg
     7201 tgtactcctc cagcgtgatg gcgctggcgg ctgccttctt cgcccggtcc tgtggagggg
     7261 tccaggtctc catctcgatg agccgcttct cgcccgcgag ccaggcttcg gcgtccatct
     7321 tgttgtcgta ggtctgcagc gcgtagtacc tcacaccgtc ctgcgggttg acgtatgagg
     7381 cttggatcct cccgctgcgc tgagtcttca gcgatcccca tccgcgacgt gccaactagg
     7441 tctcctctcg tcgtgaacaa ggctaccggg ttgcaactcc tgtgcaactc tcaggcttca
     7501 acgcgcttct acgacctgca atttctttcc acttagagga tgcagccgag agggggtaaa
     7561 aacctatctt gaccggccca tatatgtggt cggcagacac ccattcttcc aaactagcta
     7621 cgcgggttcg attcccgtcg cccgctccgc tggtcagagg gtgttttcgc cctctggcca
     7681 tttttctttc caggggtctg caactcttgt gcgactcttc tgacctgggc atacgcggtt
     7741 gcaacgcatc cctgatctgg ctactttcga tgctgacaaa cgaatagagc cccccgcctg
     7801 cgcgaacaga cgaggggcat tcacaccaga ttggagctgg tgcagtgaag agaatagacc
     7861 gggacaaggt tgcaccggga gttgcagcgg tcggaaccct cgccgtcggc gggctggcgt
     7921 tcgccctgtc gttcacggct ctagc
//
