LOCUS       sequence-313834-        7128 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bile Acid inducible sgRNA-1 with PM1 driving Nanoluc expression (NOT
            Gate 1), pNBU1 backbone, AmpR, ErmR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7128)
  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 7128)
  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: pMT448
FEATURES             Location/Qualifiers
     source          1..7128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1381..1449,1450..2241)
                     /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(1599..1618)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            2372..2481
                     /note="incP origin of transfer"
     rep_origin      2652..3040
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     misc_RNA        3273..3348
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        3527..3576
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
ORIGIN
        1 tgaacgtaga gagtagcgat attaaaagaa tccgatgaga aaagactaat atttatctat
       61 ccattcagtt tgatttctca ggactttaca tcgtcctgaa agtatttgtt ggtaccgagg
      121 acgcgtaaac atttacagtt gcatgtggcc tattgttttt agccgttaaa tattttataa
      181 ctattaaata gcgatacaaa ttgttcgaaa ctaatattgt ttatatcata tattctcgca
      241 tgttttaaag ctttattaaa ttgatttttt gtaaacagtt tttcgtactc tttgttaacc
      301 catttcatta caaaagtttc atattttttt ctctctttaa atgccatttt tgctggcttt
      361 ctttttaata caattaatgt gctatccact ttaggttttg gatggaaata atacctagga
      421 atttttgcta atatagaaat atctacctct gccattaaca gcaatgctag tgatctgttt
      481 gtatctaata acattttagc aaaaccatat tccactatta aataacttat tgtggctgaa
      541 ctttcaaaaa caatttttcg aattatattt gtgcttatgt tgtaaggtat gctgccaaat
      601 attttatatg gattgtggct aggaaatgta aatttcagta tatcatcatt tactatttga
      661 tagttaggat aatttaagag cttattacga gttacctcac ataatttaga atcaatttct
      721 atcgccgtta caaaattaca tctctttacc aatccagcag taaaatgacc tttccctgca
      781 cctatttcaa agatgttatc tttttcatct aaacttatgc aattcattat tttttctatg
      841 tgatattttg aagtaataaa attttgacta tcttttatat ttactttgtt cattataacc
      901 tctccttaat ttattgcatc tcttttcgaa tatttatgtt ttttgagaaa agaacgtact
      961 catggttcat cccgatatgc gtatcggtct gtatatcagc aactttctat gtgtttcaac
     1021 tacaatagtc atctattctc atctttctga gtccaccccc tgcaaagccc ctctttacga
     1081 cataaaaatt cggtcggaaa aggtatgcaa aagatgtttc tctctttaag agaaactctt
     1141 cgggatgcaa aaatatgaaa ataactccaa ttcaccaaat tatatagcga cttttttaca
     1201 aaatgctaaa atttgttgat ttccgtcaag caattgttga gcaaaaatgt cttttacgat
     1261 aaaatgatac ctcaatatca actgtttagc aaaacgatat ttctcttaaa gagagaaaca
     1321 cctttttgtt caccaatccc cgacttttaa tcccgcggcc atgattgaaa aaggaagagt
     1381 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     1441 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     1501 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     1561 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     1621 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     1681 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     1741 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     1801 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     1861 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     1921 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     1981 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     2041 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtca
     2101 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     2161 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     2221 tcactgatta agcattggta actgtcagac caagtttact cataacgcgt caattcgagg
     2281 gggatcaatt ccgtgatagg tgggctgccc ttcctggttg gcttggtttc atcagccatc
     2341 cgcttgccct catctgttac gccggcggta gccggccagc ctcgcagagc aggattcccg
     2401 ttgagcaccg ccaggtgcga ataagggaca gtgaagaagg aacacccgct cgcgggtggg
     2461 cctacttcac ctatcctgcc cggctgacgc cgttggatac accaaggaaa gtctacacga
     2521 accctttggc aaaatcctgt atatcgtgcg aaaaaggatg gatataccga aaaaatcgct
     2581 ataatgaccc cgaagcaggg ttatgcagcg gaaaacggaa ttgatccggc cacgatgcgt
     2641 ccggcgtaga ggatctgaag atcagcagtt caacctgttg atagtacgta ctaagctctc
     2701 atgtttcacg tactaagctc tcatgtttaa cgtactaagc tctcatgttt aacgaactaa
     2761 accctcatgg ctaacgtact aagctctcat ggctaacgta ctaagctctc atgtttcacg
     2821 tactaagctc tcatgtttga acaataaaat taatataaat cagcaactta aatagcctct
     2881 aaggttttaa gttttataag aaaaaaaaga atatataagg cttttaaagc ttttaaggtt
     2941 taacggttgt ggacaacaag ccagggatgt aacgcactga gaagccctta gagcctctca
     3001 aagcaatttt gagtgacaca ggaacactta acggctgaca tgggaattcc cctccaccgc
     3061 ggtgtattaa tgcggctgca atttttttgg gcggggccgc ccaaaaaaat cctagcaccc
     3121 tgcagggtct cccgctttac aaagaaaatt cgacaaactg ttatttttct atctatttaa
     3181 atgtacaccc gtttgtttac tttactttag ttatgtacct ttgtcggcaa tgtacacccg
     3241 aaagtttact ttcaaagata atagaactta aggttttaga gctagaaata gcaagttaaa
     3301 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt tttttggatc
     3361 caaaaaaaaa aaacacccta acgggtgttt tttttttttt ggtctgccgc ggccgctgct
     3421 gtcataggcc ggttgtccgg tagtccggat gatttaagga agatttctct tcttttttca
     3481 ctttctccga acaacctcca cttaagttct attatctttg cagtagctgt caccggatgt
     3541 gctttccggt ctgatgagtc cgtgaggacg aaacagcctc tacaaataat tttgtttaac
     3601 gcattttaaa ataaaataaa ttatttattt aattaaacga atatggtttt tactctggaa
     3661 gattttgttg gcgattggcg tcagaccgcg ggttataatt tggatcaagt cctggaacag
     3721 ggtggcgtaa gctctctgtt ccagaacctg ggtgtgagcg tgacgccgat tcagcgcatc
     3781 gttctgtccg gcgagaacgg tctgaaaatt gatattcatg tgatcatccc gtacgaaggc
     3841 ctgagcggtg accaaatggg tcaaatcgag aaaatcttta aagtcgtcta cccagttgac
     3901 gatcaccact tcaaggttat cttgcattac ggtacgctgg tgattgatgg tgtgaccccg
     3961 aatatgattg actatttcgg ccgtccgtat gaaggcattg ccgtttttga cggtaaaaag
     4021 atcaccgtca ccggtaccct gtggaatggc aataagatta ttgacgagcg tctgattaac
     4081 ccggacggca gcctgctgtt ccgcgtgacc atcaacggtg tcacgggttg gcgtctgtgc
     4141 gagcgcatcc tggcataatg aactgggatc cctcggtacc aaattttcga aaaaagacgc
     4201 tgaaaagcgt cttttttcgt tttggtccgc ggccgccggc cgctgatctg gaagaagcaa
     4261 tgaaagctgc tgttaagtct ccgaatcagg tattgttcct gacaggtgta ttcccatccg
     4321 gtaaacgcgg atactttgca gttgatctga ctcaggaata aattataaat taaggtaaga
     4381 agattgtagg ataagctaat gaaatagaaa aaggatgccg tcacacaact tgtcggcatt
     4441 cttttttgtt ttattagttg aaaatatagt gaaaaagttg cctaaatatg tatgttaaca
     4501 aattatttgt cgtaactttg cactccaaat ctgtttttaa catatggcac taatgcagaa
     4561 aaaattgact cgttctcagc agaaacactt ggacatcatc aacgctgcta aagaagaatt
     4621 catcgaattc ggtttcttgg ctgctaacat ggaccgtatc acttcttctg ctgaagtatc
     4681 taaacgtact ttgtaccgtc acttcgaatc taaagaagta ttgttcgaat ctgtattgac
     4741 tatcatcaac gactctgtaa acgaatctat ctcttaccac ttcgacccga acaaatctac
     4801 tgaagaacag ttgactgaaa tcgcttacaa agaaatcgac gtattgtaca aaacttacgg
     4861 tatcgctttg gctcgtacta tcgtaatgga attcttgcgt cagccggaaa tggctaaaac
     4921 tttgatccag aacatctact ctatccgtgc tatcactcag tggttccgtt ctgctatcga
     4981 agctaaacgt ttgaaagacg ctgacccgaa attgatgact gacgtatacg tatctttgtt
     5041 ccagggtttg ttcttctggc cgcaggtaat gcacttggac ttggaaccgc acggtgaaga
     5101 attgtctcag aaaatcgaaa ctttgactac tatcttcttg cagtcttacg gtgtagctga
     5161 ataacaatga atgggctttc ctcggtacca aattccagaa aagaggcctc ccgaaagggg
     5221 ggcctttttt cgttttggtc cctaattgcc tatcttccag tgatggaaca gcatttgtgc
     5281 attggctgca acaatcagcc ttacttgtgc ctgttctatt tccgaaccga ccgcttgtat
     5341 gaatgcatca aaattcgttt tctctacgtt ggattccttg ttgctcatat tgtgatgata
     5401 atttctacaa atatagtcat tggtaactat ctatgaaact gtttgatact tttatcagtc
     5461 cagtagtttt acaaggtctt tcttcatttc ttcgtcaata tccctgtatc gtctgaaagc
     5521 tttgcttccc tccttgtgtc ccgacagtgc ggaaacaagg ttcgggtctt tcactttttt
     5581 atagatattg ccgataaacg tacgtcttgc cagatggctg cttgccactt cataaatagg
     5641 tcgtttgatt tcgttgtgcg tcaacgggtc taagattgtt acgatgcggt caactccagc
     5701 taatttgaat atctttttta tggcatcatt gtacttttgc tcggatatga acggcaacag
     5761 ttttccctca tattctttgt agcgttcaag gatttctttc gctttgtcgt taagtggaac
     5821 acgtaccgta accggattcc cctctttggt tttcttggga atatattcta tggcttcatt
     5881 gaccacattt agtttggtca ttcggtacag gtcgctcacc ctgcatccta tcagtgtctg
     5941 aaatatgaat atatccctct gtattgccag ttgtggggtg gcagaaaggt ctgcattaaa
     6001 aatcctgtcc ctttcttcga gtgttatata ataaggtgta ccatatgtac actcctctat
     6061 cggaaacttg tcgaaaggtc tgtttgtggt gcgtttgtta tcgaagcacc acaggaagaa
     6121 tgtgcgtatt cttgaaaaac agtctatcag cgtgtttttg cttctgggct gtggtgtcct
     6181 cttttcggga atggcttcat aaatgctcgg gtaaagttca taatactggt attcgttctg
     6241 aaagaaatcc cacatatccc gaagcgtgtc aggtgttacc aaatccacat caaggataaa
     6301 gcccttttgt cctctctttg tagcccttac atatagttca taacgcagta aggctctttt
     6361 gacaacccgg aaatttttct ttcgtacttc cgacaaaggg tgcttgttta gaaattcatc
     6421 gaatagttct ccaatggtag gcttgataac cacttcctcc ggcaaaaaat atttttcagg
     6481 atggtaaaat ttatcaagtg ttgtttttag ccattctttg tctattgctt ccttttcttg
     6541 ttgatatacc ttctcgatat aggttttcaa ctggcgtatc tcttcattta tatgggtacg
     6601 catttcttca ttgcagacag ctttcgtttt tacacattcg tctttatcat cccatagatt
     6661 gggattgatg gctaactgag taggagcaac ggagtctaac tgccttccgt ttctaaaacg
     6721 gacatagatt gtagccatgg attctgtatc atatcgtttg gctgcttttt taatgataaa
     6781 ggttactttc atagactttc aggttgaatt ttactctgct gcaaatataa atattttccc
     6841 cagcattttc cccacatctg ctaaatattt tgcaattcga ttaaacttgg attaaaattt
     6901 aataggatta taacatattg aaatacagtg taattgtggc gtttttccgc atttttcttt
     6961 ttacccacat tttccccact ttaattattt caaatgcggg gtctgggtac aagaaagaaa
     7021 gctaagtatt tgatagttca atacttagct ttttcttttg cttgaatttt ccccacattt
     7081 tccccacacg tgcaaaaaat atagcagtaa gtcattattt cttttggt
//
