LOCUS       sequence-313833-        6855 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bile Acid 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 6855)
  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 6855)
  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: pMT447
FEATURES             Location/Qualifiers
     source          1..6855
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1417..1485,1486..2277)
                     /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(1635..1654)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            2408..2517
                     /note="incP origin of transfer"
     rep_origin      2688..3076
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     misc_RNA        3291..3340
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
ORIGIN
        1 caaaaaatat agcagtaagt cattatttct tttggttgaa cgtagagagt agcgatatta
       61 aaagaatccg atgagaaaag actaatattt atctatccat tcagtttgat ttctcaggac
      121 tttacatcgt cctgaaagta tttgttggta ccgaggacgc gtaaacattt acagttgcat
      181 gtggcctatt gtttttagcc gttaaatatt ttataactat taaatagcga tacaaattgt
      241 tcgaaactaa tattgtttat atcatatatt ctcgcatgtt ttaaagcttt attaaattga
      301 ttttttgtaa acagtttttc gtactctttg ttaacccatt tcattacaaa agtttcatat
      361 ttttttctct ctttaaatgc catttttgct ggctttcttt ttaatacaat taatgtgcta
      421 tccactttag gttttggatg gaaataatac ctaggaattt ttgctaatat agaaatatct
      481 acctctgcca ttaacagcaa tgctagtgat ctgtttgtat ctaataacat tttagcaaaa
      541 ccatattcca ctattaaata acttattgtg gctgaacttt caaaaacaat ttttcgaatt
      601 atatttgtgc ttatgttgta aggtatgctg ccaaatattt tatatggatt gtggctagga
      661 aatgtaaatt tcagtatatc atcatttact atttgatagt taggataatt taagagctta
      721 ttacgagtta cctcacataa tttagaatca atttctatcg ccgttacaaa attacatctc
      781 tttaccaatc cagcagtaaa atgacctttc cctgcaccta tttcaaagat gttatctttt
      841 tcatctaaac ttatgcaatt cattattttt tctatgtgat attttgaagt aataaaattt
      901 tgactatctt ttatatttac tttgttcatt ataacctctc cttaatttat tgcatctctt
      961 ttcgaatatt tatgtttttt gagaaaagaa cgtactcatg gttcatcccg atatgcgtat
     1021 cggtctgtat atcagcaact ttctatgtgt ttcaactaca atagtcatct attctcatct
     1081 ttctgagtcc accccctgca aagcccctct ttacgacata aaaattcggt cggaaaaggt
     1141 atgcaaaaga tgtttctctc tttaagagaa actcttcggg atgcaaaaat atgaaaataa
     1201 ctccaattca ccaaattata tagcgacttt tttacaaaat gctaaaattt gttgatttcc
     1261 gtcaagcaat tgttgagcaa aaatgtcttt tacgataaaa tgatacctca atatcaactg
     1321 tttagcaaaa cgatatttct cttaaagaga gaaacacctt tttgttcacc aatccccgac
     1381 ttttaatccc gcggccatga ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     1441 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     1501 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     1561 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     1621 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     1681 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     1741 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     1801 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     1861 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     1921 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     1981 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2041 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     2101 attgctgata aatctggagc cggtgagcgt gggtcacgcg gtatcattgc agcactgggg
     2161 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     2221 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     2281 tcagaccaag tttactcata acgcgtcaat tcgaggggga tcaattccgt gataggtggg
     2341 ctgcccttcc tggttggctt ggtttcatca gccatccgct tgccctcatc tgttacgccg
     2401 gcggtagccg gccagcctcg cagagcagga ttcccgttga gcaccgccag gtgcgaataa
     2461 gggacagtga agaaggaaca cccgctcgcg ggtgggccta cttcacctat cctgcccggc
     2521 tgacgccgtt ggatacacca aggaaagtct acacgaaccc tttggcaaaa tcctgtatat
     2581 cgtgcgaaaa aggatggata taccgaaaaa atcgctataa tgaccccgaa gcagggttat
     2641 gcagcggaaa acggaattga tccggccacg atgcgtccgg cgtagaggat ctgaagatca
     2701 gcagttcaac ctgttgatag tacgtactaa gctctcatgt ttcacgtact aagctctcat
     2761 gtttaacgta ctaagctctc atgtttaacg aactaaaccc tcatggctaa cgtactaagc
     2821 tctcatggct aacgtactaa gctctcatgt ttcacgtact aagctctcat gtttgaacaa
     2881 taaaattaat ataaatcagc aacttaaata gcctctaagg ttttaagttt tataagaaaa
     2941 aaaagaatat ataaggcttt taaagctttt aaggtttaac ggttgtggac aacaagccag
     3001 ggatgtaacg cactgagaag cccttagagc ctctcaaagc aattttgagt gacacaggaa
     3061 cacttaacgg ctgacatggg aattcccctc caccgcggtg tattaatgcg gctgcaattt
     3121 ttttgggcgg ggccgcccaa aaaaatccta gcaccctgca gggtctcccg ctttacaaag
     3181 aaaattcgac aaactgttat ttttctatct atttaaatgt acacccgttt gtttacttta
     3241 ctttagttat gtacctttgt cggcaatgta cacccgaaag tttactttag ctgtcaccgg
     3301 atgtgctttc cggtctgatg agtccgtgag gacgaaacag cctctacaaa taattttgtt
     3361 taacgcattt taaaataaaa taaattattt atttaattaa acgaatatgg tttttactct
     3421 ggaagatttt gttggcgatt ggcgtcagac cgcgggttat aatttggatc aagtcctgga
     3481 acagggtggc gtaagctctc tgttccagaa cctgggtgtg agcgtgacgc cgattcagcg
     3541 catcgttctg tccggcgaga acggtctgaa aattgatatt catgtgatca tcccgtacga
     3601 aggcctgagc ggtgaccaaa tgggtcaaat cgagaaaatc tttaaagtcg tctacccagt
     3661 tgacgatcac cacttcaagg ttatcttgca ttacggtacg ctggtgattg atggtgtgac
     3721 cccgaatatg attgactatt tcggccgtcc gtatgaaggc attgccgttt ttgacggtaa
     3781 aaagatcacc gtcaccggta ccctgtggaa tggcaataag attattgacg agcgtctgat
     3841 taacccggac ggcagcctgc tgttccgcgt gaccatcaac ggtgtcacgg gttggcgtct
     3901 gtgcgagcgc atcctggcat aatgaactgg gatccctcgg taccaaattt tcgaaaaaag
     3961 acgctgaaaa gcgtcttttt tcgttttggt ccgcggccgc cggccgctga tctggaagaa
     4021 gcaatgaaag ctgctgttaa gtctccgaat caggtattgt tcctgacagg tgtattccca
     4081 tccggtaaac gcggatactt tgcagttgat ctgactcagg aataaattat aaattaaggt
     4141 aagaagattg taggataagc taatgaaata gaaaaaggat gccgtcacac aacttgtcgg
     4201 cattcttttt tgttttatta gttgaaaata tagtgaaaaa gttgcctaaa tatgtatgtt
     4261 aacaaattat ttgtcgtaac tttgcactcc aaatctgttt ttaacatatg gcactaatgc
     4321 agaaaaaatt gactcgttct cagcagaaac acttggacat catcaacgct gctaaagaag
     4381 aattcatcga attcggtttc ttggctgcta acatggaccg tatcacttct tctgctgaag
     4441 tatctaaacg tactttgtac cgtcacttcg aatctaaaga agtattgttc gaatctgtat
     4501 tgactatcat caacgactct gtaaacgaat ctatctctta ccacttcgac ccgaacaaat
     4561 ctactgaaga acagttgact gaaatcgctt acaaagaaat cgacgtattg tacaaaactt
     4621 acggtatcgc tttggctcgt actatcgtaa tggaattctt gcgtcagccg gaaatggcta
     4681 aaactttgat ccagaacatc tactctatcc gtgctatcac tcagtggttc cgttctgcta
     4741 tcgaagctaa acgtttgaaa gacgctgacc cgaaattgat gactgacgta tacgtatctt
     4801 tgttccaggg tttgttcttc tggccgcagg taatgcactt ggacttggaa ccgcacggtg
     4861 aagaattgtc tcagaaaatc gaaactttga ctactatctt cttgcagtct tacggtgtag
     4921 ctgaataaca atgaatgggc tttcctcggt accaaattcc agaaaagagg cctcccgaaa
     4981 ggggggcctt ttttcgtttt ggtccctaat tgcctatctt ccagtgatgg aacagcattt
     5041 gtgcattggc tgcaacaatc agccttactt gtgcctgttc tatttccgaa ccgaccgctt
     5101 gtatgaatgc atcaaaattc gttttctcta cgttggattc cttgttgctc atattgtatg
     5161 ataatttcta caaatatagt cattggtaac tatctatgaa actgtttgat acttttatca
     5221 gtccagtagt tttacaaggt ctttcttcat ttcttcgtca atatccctgt atcgtctgaa
     5281 agctttgctt ccctccttgt gtcccgacag tgcggaaaca aggttcgggt ctttcacttt
     5341 tttatagata ttgccgataa acgtacgtct tgccagatgg ctgcttgcca cttcataaat
     5401 aggtcgtttg atttcgttgt gcgtcaacgg gtctaagatt gttacgatgc ggtcaactcc
     5461 agctaatttg aatatctttt ttatggcatc attgtacttt tgctcggata tgaacggcaa
     5521 cagttttccc tcatattctt tgtagcgttc aaggatttct ttcgctttgt cgttaagtgg
     5581 aacacgtacc gtaaccggat tcccctcttt ggttttcttg ggaatatatt ctatggcttc
     5641 attgaccaca tttagtttgg tcattcggta caggtcgctc accctgcatc ctatcagtgt
     5701 ctgaaatatg aatatatccc tctgtattgc cagttgtggg gtggcagaaa ggtctgcatt
     5761 aaaaatcctg tccctttctt cgagtgttat ataataaggt gtaccatatg tacactcctc
     5821 tatcggaaac ttgtcgaaag gtctgtttgt ggtgcgtttg ttatcgaagc accacaggaa
     5881 gaatgtgcgt attcttgaaa aacagtctat cagcgtgttt ttgcttctgg gctgtggtgt
     5941 cctcttttcg ggaatggctt cataaatgct cgggtaaagt tcataatact ggtattcgtt
     6001 ctgaaagaaa tcccacatat cccgaagcgt gtcaggtgtt accaaatcca catcaaggat
     6061 aaagcccttt tgtcctctct ttgtagccct tacatatagt tcataacgca gtaaggctct
     6121 tttgacaacc cggaaatttt tctttcgtac ttccgacaaa gggtgcttgt ttagaaattc
     6181 atcgaatagt tctccaatgg taggcttgat aaccacttcc tccggcaaaa aatatttttc
     6241 aggatggtaa aatttatcaa gtgttgtttt tagccattct ttgtctattg cttccttttc
     6301 ttgttgatat accttctcga tataggtttt caactggcgt atctcttcat ttatatgggt
     6361 acgcatttct tcattgcaga cagctttcgt ttttacacat tcgtctttat catcccatag
     6421 attgggattg atggctaact gagtaggagc aacggagtct aactgccttc cgtttctaaa
     6481 acggacatag attgtagcca tggattctgt atcatatcgt ttggctgctt ttttaatgat
     6541 aaaggttact ttcatagact ttcaggttga attttactct gctgcaaata taaatatttt
     6601 ccccagcatt ttccccacat ctgctaaata ttttgcaatt cgattaaact tggattaaaa
     6661 tttaatagga ttataacata ttgaaataca gtgtaattgt ggcgtttttc cgcatttttc
     6721 tttttaccca cattttcccc actttaatta tttcaaatgc ggggtctggg tacaagaaag
     6781 aaagctaagt atttgatagt tcaatactta gctttttctt ttgcttgaat tttccccaca
     6841 ttttccccac acgtg
//
