LOCUS       sequence-313840-        7138 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bile Acid inducible sgRNA-3 with PM3 driving Nanoluc expression (NOT
            Gate 3), pNBU1 backbone, AmpR, ErmR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7138)
  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 7138)
  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: pMT450
FEATURES             Location/Qualifiers
     source          1..7138
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        195..270
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        459..508
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             join(5441..5509,5510..6301)
                     /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(5659..5678)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            6432..6541
                     /note="incP origin of transfer"
     rep_origin      6712..7100
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 caattttttt gggcggggcc gcccaaaaaa atcctagcac cctgcagggt ctcccgcttt
       61 acaaagaaaa ttcgacaaac tgttattttt ctatctattt aaatgtacac ccgtttgttt
      121 actttacttt agttatgtac ctttgtcggc aatgtacacc cgaaagttta ctttcaaaga
      181 taattaagtt aagggtttta gagctagaaa tagcaagtta aaataaggct agtccgttat
      241 caacttgaaa aagtggcacc gagtcggtgc tttttttgga tccctcggta ccaaattcca
      301 gaaaagagac gctgaaaagc gtcttttttc gttttggtcc gcggtggctg ctgtcatagg
      361 ccggttgtcc ggtagtccgg atgatttaag gaagatttct cttctttttt cactttctcc
      421 gaacaacctc caccttaact taattatctt tgcagtagct gtcaccggat gtgctttccg
      481 gtctgatgag tccgtgagga cgaaacagcc tctacaaata attttgttta acgcatttta
      541 aaataaaata aattatttat ttaattaaac gaatatggtt tttactctgg aagattttgt
      601 tggcgattgg cgtcagaccg cgggttataa tttggatcaa gtcctggaac agggtggcgt
      661 aagctctctg ttccagaacc tgggtgtgag cgtgacgccg attcagcgca tcgttctgtc
      721 cggcgagaac ggtctgaaaa ttgatattca tgtgatcatc ccgtacgaag gcctgagcgg
      781 tgaccaaatg ggtcaaatcg agaaaatctt taaagtcgtc tacccagttg acgatcacca
      841 cttcaaggtt atcttgcatt acggtacgct ggtgattgat ggtgtgaccc cgaatatgat
      901 tgactatttc ggccgtccgt atgaaggcat tgccgttttt gacggtaaaa agatcaccgt
      961 caccggtacc ctgtggaatg gcaataagat tattgacgag cgtctgatta acccggacgg
     1021 cagcctgctg ttccgcgtga ccatcaacgg tgtcacgggt tggcgtctgt gcgagcgcat
     1081 cctggcataa tgaactggga tccctcggta ccaaattttc gaaaaaagac gctgaaaagc
     1141 gtcttttttc gttttggtcc gcggccgccg gccgctgatc tggaagaagc aatgaaagct
     1201 gctgttaagt ctccgaatca ggtattgttc ctgacaggtg tattcccatc cggtaaacgc
     1261 ggatactttg cagttgatct gactcaggaa taaattataa attaaggtaa gaagattgta
     1321 ggataagcta atgaaataga aaaaggatgc cgtcacacaa cttgtcggca ttcttttttg
     1381 ttttattagt tgaaaatata gtgaaaaagt tgcctaaata tgtatgttaa caaattattt
     1441 gtcgtaactt tgcactccaa atctgttttt aacatatggc actaatgcag aaaaaattga
     1501 ctcgttctca gcagaaacac ttggacatca tcaacgctgc taaagaagaa ttcatcgaat
     1561 tcggtttctt ggctgctaac atggaccgta tcacttcttc tgctgaagta tctaaacgta
     1621 ctttgtaccg tcacttcgaa tctaaagaag tattgttcga atctgtattg actatcatca
     1681 acgactctgt aaacgaatct atctcttacc acttcgaccc gaacaaatct actgaagaac
     1741 agttgactga aatcgcttac aaagaaatcg acgtattgta caaaacttac ggtatcgctt
     1801 tggctcgtac tatcgtaatg gaattcttgc gtcagccgga aatggctaaa actttgatcc
     1861 agaacatcta ctctatccgt gctatcactc agtggttccg ttctgctatc gaagctaaac
     1921 gtttgaaaga cgctgacccg aaattgatga ctgacgtata cgtatctttg ttccagggtt
     1981 tgttcttctg gccgcaggta atgcacttgg acttggaacc gcacggtgaa gaattgtctc
     2041 agaaaatcga aactttgact actatcttct tgcagtctta cggtgtagct gaataacaat
     2101 gaatgggctt tcctcggtac caaattccag aaaagaggcc tcccgaaagg ggggcctttt
     2161 ttcgttttgg tccctaattg cctatcttcc agtgatggaa cagcatttgt gcattggctg
     2221 caacaatcag ccttacttgt gcctgttcta tttccgaacc gaccgcttgt atgaatgcat
     2281 caaaattcgt tttctctacg ttggattcct tgttgctcat attgtgatga taatttctac
     2341 aaatatagtc attggtaact atctatgaaa ctgtttgata cttttatcag tccagtagtt
     2401 ttacaaggtc tttcttcatt tcttcgtcaa tatccctgta tcgtctgaaa gctttgcttc
     2461 cctccttgtg tcccgacagt gcggaaacaa ggttcgggtc tttcactttt ttatagatat
     2521 tgccgataaa cgtacgtctt gccagatggc tgcttgccac ttcataaata ggtcgtttga
     2581 tttcgttgtg cgtcaacggg tctaagattg ttacgatgcg gtcaactcca gctaatttga
     2641 atatcttttt tatggcatca ttgtactttt gctcggatat gaacggcaac agttttccct
     2701 catattcttt gtagcgttca aggatttctt tcgctttgtc gttaagtgga acacgtaccg
     2761 taaccggatt cccctctttg gttttcttgg gaatatattc tatggcttca ttgaccacat
     2821 ttagtttggt cattcggtac aggtcgctca ccctgcatcc tatcagtgtc tgaaatatga
     2881 atatatccct ctgtattgcc agttgtgggg tggcagaaag gtctgcatta aaaatcctgt
     2941 ccctttcttc gagtgttata taataaggtg taccatatgt acactcctct atcggaaact
     3001 tgtcgaaagg tctgtttgtg gtgcgtttgt tatcgaagca ccacaggaag aatgtgcgta
     3061 ttcttgaaaa acagtctatc agcgtgtttt tgcttctggg ctgtggtgtc ctcttttcgg
     3121 gaatggcttc ataaatgctc gggtaaagtt cataatactg gtattcgttc tgaaagaaat
     3181 cccacatatc ccgaagcgtg tcaggtgtta ccaaatccac atcaaggata aagccctttt
     3241 gtcctctctt tgtagccctt acatatagtt cataacgcag taaggctctt ttgacaaccc
     3301 ggaaattttt ctttcgtact tccgacaaag ggtgcttgtt tagaaattca tcgaatagtt
     3361 ctccaatggt aggcttgata accacttcct ccggcaaaaa atatttttca ggatggtaaa
     3421 atttatcaag tgttgttttt agccattctt tgtctattgc ttccttttct tgttgatata
     3481 ccttctcgat ataggttttc aactggcgta tctcttcatt tatatgggta cgcatttctt
     3541 cattgcagac agctttcgtt tttacacatt cgtctttatc atcccataga ttgggattga
     3601 tggctaactg agtaggagca acggagtcta actgccttcc gtttctaaaa cggacataga
     3661 ttgtagccat ggattctgta tcatatcgtt tggctgcttt tttaatgata aaggttactt
     3721 tcatagactt tcaggttgaa ttttactctg ctgcaaatat aaatattttc cccagcattt
     3781 tccccacatc tgctaaatat tttgcaattc gattaaactt ggattaaaat ttaataggat
     3841 tataacatat tgaaatacag tgtaattgtg gcgtttttcc gcatttttct ttttacccac
     3901 attttcccca ctttaattat ttcaaatgcg gggtctgggt acaagaaaga aagctaagta
     3961 tttgatagtt caatacttag ctttttcttt tgcttgaatt ttccccacat tttccccaca
     4021 cgtgcaaaaa atatagcagt aagtcattat ttcttttggt tgaacgtaga gagtagcgat
     4081 attaaaagaa tccgatgaga aaagactaat atttatctat ccattcagtt tgatttctca
     4141 ggactttaca tcgtcctgaa agtatttgtt ggtaccgagg acgcgtaaac atttacagtt
     4201 gcatgtggcc tattgttttt agccgttaaa tattttataa ctattaaata gcgatacaaa
     4261 ttgttcgaaa ctaatattgt ttatatcata tattctcgca tgttttaaag ctttattaaa
     4321 ttgatttttt gtaaacagtt tttcgtactc tttgttaacc catttcatta caaaagtttc
     4381 atattttttt ctctctttaa atgccatttt tgctggcttt ctttttaata caattaatgt
     4441 gctatccact ttaggttttg gatggaaata atacctagga atttttgcta atatagaaat
     4501 atctacctct gccattaaca gcaatgctag tgatctgttt gtatctaata acattttagc
     4561 aaaaccatat tccactatta aataacttat tgtggctgaa ctttcaaaaa caatttttcg
     4621 aattatattt gtgcttatgt tgtaaggtat gctgccaaat attttatatg gattgtggct
     4681 aggaaatgta aatttcagta tatcatcatt tactatttga tagttaggat aatttaagag
     4741 cttattacga gttacctcac ataatttaga atcaatttct atcgccgtta caaaattaca
     4801 tctctttacc aatccagcag taaaatgacc tttccctgca cctatttcaa agatgttatc
     4861 tttttcatct aaacttatgc aattcattat tttttctatg tgatattttg aagtaataaa
     4921 attttgacta tcttttatat ttactttgtt cattataacc tctccttaat ttattgcatc
     4981 tcttttcgaa tatttatgtt ttttgagaaa agaacgtact catggttcat cccgatatgc
     5041 gtatcggtct gtatatcagc aactttctat gtgtttcaac tacaatagtc atctattctc
     5101 atctttctga gtccaccccc tgcaaagccc ctctttacga cataaaaatt cggtcggaaa
     5161 aggtatgcaa aagatgtttc tctctttaag agaaactctt cgggatgcaa aaatatgaaa
     5221 ataactccaa ttcaccaaat tatatagcga cttttttaca aaatgctaaa atttgttgat
     5281 ttccgtcaag caattgttga gcaaaaatgt cttttacgat aaaatgatac ctcaatatca
     5341 actgtttagc aaaacgatat ttctcttaaa gagagaaaca cctttttgtt caccaatccc
     5401 cgacttttaa tcccgcggcc atgattgaaa aaggaagagt atgagtattc aacatttccg
     5461 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5521 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5581 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5641 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5701 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5761 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5821 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5881 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5941 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6001 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6061 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6121 gtttattgct gataaatctg gagccggtga gcgtgggtca cgcggtatca ttgcagcact
     6181 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6241 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6301 actgtcagac caagtttact cataacgcgt caattcgagg gggatcaatt ccgtgatagg
     6361 tgggctgccc ttcctggttg gcttggtttc atcagccatc cgcttgccct catctgttac
     6421 gccggcggta gccggccagc ctcgcagagc aggattcccg ttgagcaccg ccaggtgcga
     6481 ataagggaca gtgaagaagg aacacccgct cgcgggtggg cctacttcac ctatcctgcc
     6541 cggctgacgc cgttggatac accaaggaaa gtctacacga accctttggc aaaatcctgt
     6601 atatcgtgcg aaaaaggatg gatataccga aaaaatcgct ataatgaccc cgaagcaggg
     6661 ttatgcagcg gaaaacggaa ttgatccggc cacgatgcgt ccggcgtaga ggatctgaag
     6721 atcagcagtt caacctgttg atagtacgta ctaagctctc atgtttcacg tactaagctc
     6781 tcatgtttaa cgtactaagc tctcatgttt aacgaactaa accctcatgg ctaacgtact
     6841 aagctctcat ggctaacgta ctaagctctc atgtttcacg tactaagctc tcatgtttga
     6901 acaataaaat taatataaat cagcaactta aatagcctct aaggttttaa gttttataag
     6961 aaaaaaaaga atatataagg cttttaaagc ttttaaggtt taacggttgt ggacaacaag
     7021 ccagggatgt aacgcactga gaagccctta gagcctctca aagcaatttt gagtgacaca
     7081 ggaacactta acggctgaca tgggaattcc cctccaccgc ggtgtattaa tgcggctg
//
