LOCUS       sequence-313827-        5907 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  aTC inducible Nanoluc expression, pNBU1 backbone with BsaI sites, 
            AmpR, ErmR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5907)
  AUTHORS   Taketani M, Zhang J, Zhang S, Triassi AJ, Huang YJ, Griffith LG, 
            Voigt CA
  TITLE     Genetic circuit design automation for the gut resident species 
            Bacteroides thetaiotaomicron.
  JOURNAL   Nat Biotechnol. 2020 Aug;38(8):962-969. doi: 
            10.1038/s41587-020-0468-5. Epub 2020 Mar 30.
  PUBMED    32231334
REFERENCE   2  (bases 1 to 5907)
  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: 
            "10.1038/s41587-020-0468-5"; journalName: "Nat Biotechnol"; date: 
            "2020-08"; volume: "38"; issue: "8"; pages: "962-969"
COMMENT     Sequence Label: pMT445
FEATURES             Location/Qualifiers
     source          1..5907
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    78..96
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    140..158
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        165..214
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             join(4141..4209,4210..5001)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4359..4378)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            5132..5241
                     /note="incP origin of transfer"
     rep_origin      5412..5800
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 ctttccaaga gaagaaagcc ttgttaaatt gacttagtgt aaaagcgcag tactgcttga
       61 ccataagaac aaaaaaatct ctatcactga tagggataaa gtttggaaga taaagctaaa
      121 agttcttatc tttgcagtct ccctatcagt gatagagacg caagctgtca ccggatgtgc
      181 tttccggtct gatgagtccg tgaggacgaa acagcctcta caaataattt tgtttaacgc
      241 attttaaaat aaaataaatt atttatttaa ttaaacgaat atggttttta ctctggaaga
      301 ttttgttggc gattggcgtc agaccgcggg ttataatttg gatcaagtcc tggaacaggg
      361 tggcgtaagc tctctgttcc agaacctggg tgtgagcgtg acgccgattc agcgcatcgt
      421 tctgtccggc gagaacggtc tgaaaattga tattcatgtg atcatcccgt acgaaggcct
      481 gagcggtgac caaatgggtc aaatcgagaa aatctttaaa gtcgtctacc cagttgacga
      541 tcaccacttc aaggttatct tgcattacgg tacgctggtg attgatggtg tgaccccgaa
      601 tatgattgac tatttcggcc gtccgtatga aggcattgcc gtttttgacg gtaaaaagat
      661 caccgtcacc ggtaccctgt ggaatggcaa taagattatt gacgagcgtc tgattaaccc
      721 ggacggcagc ctgctgttcc gcgtgaccat caacggtgtc acgggttggc gtctgtgcga
      781 gcgcatcctg gcataacaat gaatgggctt tcctcggtac caaattccag aaaagaggcc
      841 tcccgaaagg ggggcctttt ttcgttttgg tccctaattg cctatcttcc agtgatggaa
      901 cagcatttgt gcattggctg caacaatcag ccttacttgt gcctgttcta tttccgaacc
      961 gaccgcttgt atgaatgcat caaaattcgt tttctctacg ttggattcct tgttgctcat
     1021 attgtgatga taatttctac aaatatagtc attggtaact atctatgaaa ctgtttgata
     1081 cttttatcag tccagtagtt ttacaaggtc tttcttcatt tcttcgtcaa tatccctgta
     1141 tcgtctgaaa gctttgcttc cctccttgtg tcccgacagt gcggaaacaa ggttcgggtc
     1201 tttcactttt ttatagatat tgccgataaa cgtacgtctt gccagatggc tgcttgccac
     1261 ttcataaata ggtcgtttga tttcgttgtg cgtcaacggg tctaagattg ttacgatgcg
     1321 gtcaactcca gctaatttga atatcttttt tatggcatca ttgtactttt gctcggatat
     1381 gaacggcaac agttttccct catattcttt gtagcgttca aggatttctt tcgctttgtc
     1441 gttaagtgga acacgtaccg taaccggatt cccctctttg gttttcttgg gaatatattc
     1501 tatggcttca ttgaccacat ttagtttggt cattcggtac aggtcgctca ccctgcatcc
     1561 tatcagtgtc tgaaatatga atatatccct ctgtattgcc agttgtgggg tggcagaaag
     1621 gtctgcatta aaaatcctgt ccctttcttc gagtgttata taataaggtg taccatatgt
     1681 acactcctct atcggaaact tgtcgaaagg tctgtttgtg gtgcgtttgt tatcgaagca
     1741 ccacaggaag aatgtgcgta ttcttgaaaa acagtctatc agcgtgtttt tgcttctggg
     1801 ctgtggtgtc ctcttttcgg gaatggcttc ataaatgctc gggtaaagtt cataatactg
     1861 gtattcgttc tgaaagaaat cccacatatc ccgaagcgtg tcaggtgtta ccaaatccac
     1921 atcaaggata aagccctttt gtcctctctt tgtagccctt acatatagtt cataacgcag
     1981 taaggctctt ttgacaaccc ggaaattttt ctttcgtact tccgacaaag ggtgcttgtt
     2041 tagaaattca tcgaatagtt ctccaatggt aggcttgata accacttcct ccggcaaaaa
     2101 atatttttca ggatggtaaa atttatcaag tgttgttttt agccattctt tgtctattgc
     2161 ttccttttct tgttgatata ccttctcgat ataggttttc aactggcgta tctcttcatt
     2221 tatatgggta cgcatttctt cattgcagac agctttcgtt tttacacatt cgtctttatc
     2281 atcccataga ttgggattga tggctaactg agtaggagca acggagtcta actgccttcc
     2341 gtttctaaaa cggacataga ttgtagccat ggattctgta tcatatcgtt tggctgcttt
     2401 tttaatgata aaggttactt tcatagactt tcaggttgaa ttttactctg ctgcaaatat
     2461 aaatattttc cccagcattt tccccacatc tgctaaatat tttgcaattc gattaaactt
     2521 ggattaaaat ttaataggat tataacatat tgaaatacag tgtaattgtg gcgtttttcc
     2581 gcatttttct ttttacccac attttcccca ctttaattat ttcaaatgcg gggtctgggt
     2641 acaagaaaga aagctaagta tttgatagtt caatacttag ctttttcttt tgcttgaatt
     2701 ttccccacat tttccccaca cgtgcaaaaa atatagcagt aagtcattat ttcttttggt
     2761 tgaacgtaga gagtagcgat attaaaagaa tccgatgaga aaagactaat atttatctat
     2821 ccattcagtt tgatttctca ggactttaca tcgtcctgaa agtatttgtt ggtaccgagg
     2881 acgcgtaaac atttacagtt gcatgtggcc tattgttttt agccgttaaa tattttataa
     2941 ctattaaata gcgatacaaa ttgttcgaaa ctaatattgt ttatatcata tattctcgca
     3001 tgttttaaag ctttattaaa ttgatttttt gtaaacagtt tttcgtactc tttgttaacc
     3061 catttcatta caaaagtttc atattttttt ctctctttaa atgccatttt tgctggcttt
     3121 ctttttaata caattaatgt gctatccact ttaggttttg gatggaaata atacctagga
     3181 atttttgcta atatagaaat atctacctct gccattaaca gcaatgctag tgatctgttt
     3241 gtatctaata acattttagc aaaaccatat tccactatta aataacttat tgtggctgaa
     3301 ctttcaaaaa caatttttcg aattatattt gtgcttatgt tgtaaggtat gctgccaaat
     3361 attttatatg gattgtggct aggaaatgta aatttcagta tatcatcatt tactatttga
     3421 tagttaggat aatttaagag cttattacga gttacctcac ataatttaga atcaatttct
     3481 atcgccgtta caaaattaca tctctttacc aatccagcag taaaatgacc tttccctgca
     3541 cctatttcaa agatgttatc tttttcatct aaacttatgc aattcattat tttttctatg
     3601 tgatattttg aagtaataaa attttgacta tcttttatat ttactttgtt cattataacc
     3661 tctccttaat ttattgcatc tcttttcgaa tatttatgtt ttttgagaaa agaacgtact
     3721 catggttcat cccgatatgc gtatcggtct gtatatcagc aactttctat gtgtttcaac
     3781 tacaatagtc atctattctc atctttctga gtccaccccc tgcaaagccc ctctttacga
     3841 cataaaaatt cggtcggaaa aggtatgcaa aagatgtttc tctctttaag agaaactctt
     3901 cgggatgcaa aaatatgaaa ataactccaa ttcaccaaat tatatagcga cttttttaca
     3961 aaatgctaaa atttgttgat ttccgtcaag caattgttga gcaaaaatgt cttttacgat
     4021 aaaatgatac ctcaatatca actgtttagc aaaacgatat ttctcttaaa gagagaaaca
     4081 cctttttgtt caccaatccc cgacttttaa tcccgcggcc atgattgaaa aaggaagagt
     4141 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     4201 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     4261 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     4321 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     4381 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     4441 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     4501 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     4561 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     4621 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     4681 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     4741 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     4801 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtca
     4861 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     4921 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4981 tcactgatta agcattggta actgtcagac caagtttact cataacgcgt caattcgagg
     5041 gggatcaatt ccgtgatagg tgggctgccc ttcctggttg gcttggtttc atcagccatc
     5101 cgcttgccct catctgttac gccggcggta gccggccagc ctcgcagagc aggattcccg
     5161 ttgagcaccg ccaggtgcga ataagggaca gtgaagaagg aacacccgct cgcgggtggg
     5221 cctacttcac ctatcctgcc cggctgacgc cgttggatac accaaggaaa gtctacacga
     5281 accctttggc aaaatcctgt atatcgtgcg aaaaaggatg gatataccga aaaaatcgct
     5341 ataatgaccc cgaagcaggg ttatgcagcg gaaaacggaa ttgatccggc cacgatgcgt
     5401 ccggcgtaga ggatctgaag atcagcagtt caacctgttg atagtacgta ctaagctctc
     5461 atgtttcacg tactaagctc tcatgtttaa cgtactaagc tctcatgttt aacgaactaa
     5521 accctcatgg ctaacgtact aagctctcat ggctaacgta ctaagctctc atgtttcacg
     5581 tactaagctc tcatgtttga acaataaaat taatataaat cagcaactta aatagcctct
     5641 aaggttttaa gttttataag aaaaaaaaga atatataagg cttttaaagc ttttaaggtt
     5701 taacggttgt ggacaacaag ccagggatgt aacgcactga gaagccctta gagcctctca
     5761 aagcaatttt gagtgacaca ggaacactta acggctgaca tgggaattcc cctccaccgc
     5821 ggtgtattaa tgcggctgca atttttttgg gcggggccgc ccaaaaaaat cctagcaccc
     5881 tgcagggtct cccgcttttt gcacccg
//
