LOCUS       sequence-313820-        5877 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG 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 5877)
  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 5877)
  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: pMT444
FEATURES             Location/Qualifiers
     source          1..5877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        complement(602..651)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     protein_bind    698..714
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    731..747
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     rep_origin      893..1281
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            1452..1561
                     /note="incP origin of transfer"
     CDS             complement(join(1692..2483,2484..2552))
                     /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     2315..2334
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gaggaaagcc cattcattgt tatgccagga tgcgctcgca cagacgccaa cccgtgacac
       61 cgttgatggt cacgcggaac agcaggctgc cgtccgggtt aatcagacgc tcgtcaataa
      121 tcttattgcc attccacagg gtaccggtga cggtgatctt tttaccgtca aaaacggcaa
      181 tgccttcata cggacggccg aaatagtcaa tcatattcgg ggtcacacca tcaatcacca
      241 gcgtaccgta atgcaagata accttgaagt ggtgatcgtc aactgggtag acgactttaa
      301 agattttctc gatttgaccc atttggtcac cgctcaggcc ttcgtacggg atgatcacat
      361 gaatatcaat tttcagaccg ttctcgccgg acagaacgat gcgctgaatc ggcgtcacgc
      421 tcacacccag gttctggaac agagagctta cgccaccctg ttccaggact tgatccaaat
      481 tataacccgc ggtctgacgc caatcgccaa caaaatcttc cagagtaaaa accatattcg
      541 tttaattaaa taaataattt attttatttt aaaatgcgtt aaacaaaatt atttgtagag
      601 gctgtttcgt cctcacggac tcatcagacc ggaaagcaca tccggtgaca gctgtttata
      661 ttatttatat ttgtttgacg agaatatctt tatttaattg ttatccgctc acaattgccg
      721 acaaaggtaa ttgttatccg ctcacaattc aaataaatag atagaaaaat aacagtttgt
      781 cgaattttct ttgtaaagcg ggagaccctg cagggtgcta ggattttttt gggcggcccc
      841 gcccaaaaaa attgcagccg cattaataca ccgcggtgga ggggaattcc catgtcagcc
      901 gttaagtgtt cctgtgtcac tcaaaattgc tttgagaggc tctaagggct tctcagtgcg
      961 ttacatccct ggcttgttgt ccacaaccgt taaaccttaa aagctttaaa agccttatat
     1021 attctttttt ttcttataaa acttaaaacc ttagaggcta tttaagttgc tgatttatat
     1081 taattttatt gttcaaacat gagagcttag tacgtgaaac atgagagctt agtacgttag
     1141 ccatgagagc ttagtacgtt agccatgagg gtttagttcg ttaaacatga gagcttagta
     1201 cgttaaacat gagagcttag tacgtgaaac atgagagctt agtacgtact atcaacaggt
     1261 tgaactgctg atcttcagat cctctacgcc ggacgcatcg tggccggatc aattccgttt
     1321 tccgctgcat aaccctgctt cggggtcatt atagcgattt tttcggtata tccatccttt
     1381 ttcgcacgat atacaggatt ttgccaaagg gttcgtgtag actttccttg gtgtatccaa
     1441 cggcgtcagc cgggcaggat aggtgaagta ggcccacccg cgagcgggtg ttccttcttc
     1501 actgtccctt attcgcacct ggcggtgctc aacgggaatc ctgctctgcg aggctggccg
     1561 gctaccgccg gcgtaacaga tgagggcaag cggatggctg atgaaaccaa gccaaccagg
     1621 aagggcagcc cacctatcac ggaattgatc cccctcgaat tgacgcgtta tgagtaaact
     1681 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     1741 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     1801 ccatctggcc ccagtgctgc aatgataccg cgtgacccac gctcaccggc tccagattta
     1861 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     1921 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     1981 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     2041 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     2101 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     2161 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2221 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2281 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     2341 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     2401 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     2461 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     2521 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaatc atggccgcgg
     2581 gattaaaagt cggggattgg tgaacaaaaa ggtgtttctc tctttaagag aaatatcgtt
     2641 ttgctaaaca gttgatattg aggtatcatt ttatcgtaaa agacattttt gctcaacaat
     2701 tgcttgacgg aaatcaacaa attttagcat tttgtaaaaa agtcgctata taatttggtg
     2761 aattggagtt attttcatat ttttgcatcc cgaagagttt ctcttaaaga gagaaacatc
     2821 ttttgcatac cttttccgac cgaattttta tgtcgtaaag aggggctttg cagggggtgg
     2881 actcagaaag atgagaatag atgactattg tagttgaaac acatagaaag ttgctgatat
     2941 acagaccgat acgcatatcg ggatgaacca tgagtacgtt cttttctcaa aaaacataaa
     3001 tattcgaaaa gagatgcaat aaattaagga gaggttataa tgaacaaagt aaatataaaa
     3061 gatagtcaaa attttattac ttcaaaatat cacatagaaa aaataatgaa ttgcataagt
     3121 ttagatgaaa aagataacat ctttgaaata ggtgcaggga aaggtcattt tactgctgga
     3181 ttggtaaaga gatgtaattt tgtaacggcg atagaaattg attctaaatt atgtgaggta
     3241 actcgtaata agctcttaaa ttatcctaac tatcaaatag taaatgatga tatactgaaa
     3301 tttacatttc ctagccacaa tccatataaa atatttggca gcatacctta caacataagc
     3361 acaaatataa ttcgaaaaat tgtttttgaa agttcagcca caataagtta tttaatagtg
     3421 gaatatggtt ttgctaaaat gttattagat acaaacagat cactagcatt gctgttaatg
     3481 gcagaggtag atatttctat attagcaaaa attcctaggt attatttcca tccaaaacct
     3541 aaagtggata gcacattaat tgtattaaaa agaaagccag caaaaatggc atttaaagag
     3601 agaaaaaaat atgaaacttt tgtaatgaaa tgggttaaca aagagtacga aaaactgttt
     3661 acaaaaaatc aatttaataa agctttaaaa catgcgagaa tatatgatat aaacaatatt
     3721 agtttcgaac aatttgtatc gctatttaat agttataaaa tatttaacgg ctaaaaacaa
     3781 taggccacat gcaactgtaa atgtttacgc gtcctcggta ccaacaaata ctttcaggac
     3841 gatgtaaagt cctgagaaat caaactgaat ggatagataa atattagtct tttctcatcg
     3901 gattctttta atatcgctac tctctacgtt caaccaaaag aaataatgac ttactgctat
     3961 attttttgca cgtgtgggga aaatgtgggg aaaattcaag caaaagaaaa agctaagtat
     4021 tgaactatca aatacttagc tttctttctt gtacccagac cccgcatttg aaataattaa
     4081 agtggggaaa atgtgggtaa aaagaaaaat gcggaaaaac gccacaatta cactgtattt
     4141 caatatgtta taatcctatt aaattttaat ccaagtttaa tcgaattgca aaatatttag
     4201 cagatgtggg gaaaatgctg gggaaaatat ttatatttgc agcagagtaa aattcaacct
     4261 gaaagtctat gaaagtaacc tttatcatta aaaaagcagc caaacgatat gatacagaat
     4321 ccatggctac aatctatgtc cgttttagaa acggaaggca gttagactcc gttgctccta
     4381 ctcagttagc catcaatccc aatctatggg atgataaaga cgaatgtgta aaaacgaaag
     4441 ctgtctgcaa tgaagaaatg cgtacccata taaatgaaga gatacgccag ttgaaaacct
     4501 atatcgagaa ggtatatcaa caagaaaagg aagcaataga caaagaatgg ctaaaaacaa
     4561 cacttgataa attttaccat cctgaaaaat attttttgcc ggaggaagtg gttatcaagc
     4621 ctaccattgg agaactattc gatgaatttc taaacaagca ccctttgtcg gaagtacgaa
     4681 agaaaaattt ccgggttgtc aaaagagcct tactgcgtta tgaactatat gtaagggcta
     4741 caaagagagg acaaaagggc tttatccttg atgtggattt ggtaacacct gacacgcttc
     4801 gggatatgtg ggatttcttt cagaacgaat accagtatta tgaactttac ccgagcattt
     4861 atgaagccat tcccgaaaag aggacaccac agcccagaag caaaaacacg ctgatagact
     4921 gtttttcaag aatacgcaca ttcttcctgt ggtgcttcga taacaaacgc accacaaaca
     4981 gacctttcga caagtttccg atagaggagt gtacatatgg tacaccttat tatataacac
     5041 tcgaagaaag ggacaggatt tttaatgcag acctttctgc caccccacaa ctggcaatac
     5101 agagggatat attcatattt cagacactga taggatgcag ggtgagcgac ctgtaccgaa
     5161 tgaccaaact aaatgtggtc aatgaagcca tagaatatat tcccaagaaa accaaagagg
     5221 ggaatccggt tacggtacgt gttccactta acgacaaagc gaaagaaatc cttgaacgct
     5281 acaaagaata tgagggaaaa ctgttgccgt tcatatccga gcaaaagtac aatgatgcca
     5341 taaaaaagat attcaaatta gctggagttg accgcatcgt aacaatctta gacccgttga
     5401 cgcacaacga aatcaaacga cctatttatg aagtggcaag cagccatctg gcaagacgta
     5461 cgtttatcgg caatatctat aaaaaagtga aagacccgaa ccttgtttcc gcactgtcgg
     5521 gacacaagga gggaagcaaa gctttcagac gatacaggga tattgacgaa gaaatgaaga
     5581 aagaccttgt aaaactactg gactgataaa agtatcaaac agtttcatag atagttacca
     5641 atgactatat ttgtagaaat tatcatcaca atatgagcaa caaggaatcc aacgtagaga
     5701 aaacgaattt tgatgcattc atacaagcgg tcggttcgga aatagaacag gcacaagtaa
     5761 ggctgattgt tgcagccaat gcacaaatgc tgttccatca ctggaagata ggcaattagg
     5821 gaccaaaacg aaaaaaggcc cccctttcgg gaggcctctt ttctggaatt tggtacc
//
