LOCUS       sequence-313938-        7125 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bile Acid inducible sgRNA-5 with PM5 driving Nanoluc expression (NOT
            Gate 5), pNBU1 backbone, AmpR, ErmR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7125)
  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 7125)
  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: pMT492
FEATURES             Location/Qualifiers
     source          1..7125
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1345..1413,1414..2205)
                     /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(1563..1582)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            2336..2445
                     /note="incP origin of transfer"
     rep_origin      2616..3004
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     misc_RNA        3237..3312
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        3488..3537
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
ORIGIN
        1 gagaaaagac taatatttat ctatccattc agtttgattt ctcaggactt tacatcgtcc
       61 tgaaagtatt tgttggtacc gaggacgcgt aaacatttac agttgcatgt ggcctattgt
      121 ttttagccgt taaatatttt ataactatta aatagcgata caaattgttc gaaactaata
      181 ttgtttatat catatattct cgcatgtttt aaagctttat taaattgatt ttttgtaaac
      241 agtttttcgt actctttgtt aacccatttc attacaaaag tttcatattt ttttctctct
      301 ttaaatgcca tttttgctgg ctttcttttt aatacaatta atgtgctatc cactttaggt
      361 tttggatgga aataatacct aggaattttt gctaatatag aaatatctac ctctgccatt
      421 aacagcaatg ctagtgatct gtttgtatct aataacattt tagcaaaacc atattccact
      481 attaaataac ttattgtggc tgaactttca aaaacaattt ttcgaattat atttgtgctt
      541 atgttgtaag gtatgctgcc aaatatttta tatggattgt ggctaggaaa tgtaaatttc
      601 agtatatcat catttactat ttgatagtta ggataattta agagcttatt acgagttacc
      661 tcacataatt tagaatcaat ttctatcgcc gttacaaaat tacatctctt taccaatcca
      721 gcagtaaaat gacctttccc tgcacctatt tcaaagatgt tatctttttc atctaaactt
      781 atgcaattca ttattttttc tatgtgatat tttgaagtaa taaaattttg actatctttt
      841 atatttactt tgttcattat aacctctcct taatttattg catctctttt cgaatattta
      901 tgttttttga gaaaagaacg tactcatggt tcatcccgat atgcgtatcg gtctgtatat
      961 cagcaacttt ctatgtgttt caactacaat agtcatctat tctcatcttt ctgagtccac
     1021 cccctgcaaa gcccctcttt acgacataaa aattcggtcg gaaaaggtat gcaaaagatg
     1081 tttctctctt taagagaaac tcttcgggat gcaaaaatat gaaaataact ccaattcacc
     1141 aaattatata gcgacttttt tacaaaatgc taaaatttgt tgatttccgt caagcaattg
     1201 ttgagcaaaa atgtctttta cgataaaatg atacctcaat atcaactgtt tagcaaaacg
     1261 atatttctct taaagagaga aacacctttt tgttcaccaa tccccgactt ttaatcccgc
     1321 ggccatgatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     1381 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     1441 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     1501 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     1561 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     1621 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     1681 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     1741 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     1801 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     1861 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     1921 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     1981 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     2041 tctggagccg gtgagcgtgg gtcacgcggt atcattgcag cactggggcc agatggtaag
     2101 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     2161 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     2221 tactcataac gcgtcaattc gagggggatc aattccgtga taggtgggct gcccttcctg
     2281 gttggcttgg tttcatcagc catccgcttg ccctcatctg ttacgccggc ggtagccggc
     2341 cagcctcgca gagcaggatt cccgttgagc accgccaggt gcgaataagg gacagtgaag
     2401 aaggaacacc cgctcgcggg tgggcctact tcacctatcc tgcccggctg acgccgttgg
     2461 atacaccaag gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag
     2521 gatggatata ccgaaaaaat cgctataatg accccgaagc agggttatgc agcggaaaac
     2581 ggaattgatc cggccacgat gcgtccggcg tagaggatct gaagatcaac agttcaacct
     2641 gttgatagta cgtactaagc tctcatgttt cacgtactaa gctctcatgt ttaacgtact
     2701 aagctctcat gtttaacgaa ctaaaccctc atggctaacg tactaagctc tcatggctaa
     2761 cgtactaagc tctcatgttt cacgtactaa gctctcatgt ttgaacaata aaattaatat
     2821 aaatcagcaa cttaaatagc ctctaaggtt ttaagtttta taagaaaaaa aagaatatat
     2881 aaggctttta aagcttttaa ggtttaacgg ttgtggacaa caagccaggg atgtaacgca
     2941 ctgagaagcc cttagagcct ctcaaagcaa ttttgagtga cacaggaaca cttaacggct
     3001 gacatgggaa ttcccctcca ccgcggtgta ttaatgcggc tgcaattttt ttgggcgggg
     3061 ccgcccaaaa aaatcctagc accctgcagg gtctcccgct ttacaaagaa aattcgacaa
     3121 actgttattt ttctatctat ttaaatgtac acccgtttgt ttactttact ttagttatgt
     3181 acctttgtcg gcaatgtaca cccgaaagtt tactttcaaa gatagaataa gtctttgttt
     3241 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca
     3301 ccgagtcggt gctttttttc caattattga acaccctaac gggtgttttt ttgtttctgg
     3361 tctcccgccg cggccgctgc tgtcataggc cggttgtccg gtagtccgga tgatttaagg
     3421 aagatttctc ttcttttttc actttctccg aacaacctcc aaaagactta ttctatcttt
     3481 gcagtagctg tcaccggatg tgctttccgg tctgatgagt ccgtgaggac gaaacagcct
     3541 ctacaaataa ttttgtttaa cgcattttaa aataaaataa attatttatt taattaaacg
     3601 aatatggttt ttactctgga agattttgtt ggcgattggc gtcagaccgc gggttataat
     3661 ttggatcaag tcctggaaca gggtggcgta agctctctgt tccagaacct gggtgtgagc
     3721 gtgacgccga ttcagcgcat cgttctgtcc ggcgagaacg gtctgaaaat tgatattcat
     3781 gtgatcatcc cgtacgaagg cctgagcggt gaccaaatgg gtcaaatcga gaaaatcttt
     3841 aaagtcgtct acccagttga cgatcaccac ttcaaggtta tcttgcatta cggtacgctg
     3901 gtgattgatg gtgtgacccc gaatatgatt gactatttcg gccgtccgta tgaaggcatt
     3961 gccgtttttg acggtaaaaa gatcaccgtc accggtaccc tgtggaatgg caataagatt
     4021 attgacgagc gtctgattaa cccggacggc agcctgctgt tccgcgtgac catcaacggt
     4081 gtcacgggtt ggcgtctgtg cgagcgcatc ctggcataat gaactgggat ccctcggtac
     4141 caaattttcg aaaaaagacg ctgaaaagcg tcttttttcg ttttggtccg cggccgccgg
     4201 ccgctgatct ggaagaagca atgaaagctg ctgttaagtc tccgaatcag gtattgttcc
     4261 tgacaggtgt attcccatcc ggtaaacgcg gatactttgc agttgatctg actcaggaat
     4321 aaattataaa ttaaggtaag aagattgtag gataagctaa tgaaatagaa aaaggatgcc
     4381 gtcacacaac ttgtcggcat tcttttttgt tttattagtt gaaaatatag tgaaaaagtt
     4441 gcctaaatat gtatgttaac aaattatttg tcgtaacttt gcactccaaa tctgttttta
     4501 acatatggca ctaatgcaga aaaaattgac tcgttctcag cagaaacact tggacatcat
     4561 caacgctgct aaagaagaat tcatcgaatt cggtttcttg gctgctaaca tggaccgtat
     4621 cacttcttct gctgaagtat ctaaacgtac tttgtaccgt cacttcgaat ctaaagaagt
     4681 attgttcgaa tctgtattga ctatcatcaa cgactctgta aacgaatcta tctcttacca
     4741 cttcgacccg aacaaatcta ctgaagaaca gttgactgaa atcgcttaca aagaaatcga
     4801 cgtattgtac aaaacttacg gtatcgcttt ggctcgtact atcgtaatgg aattcttgcg
     4861 tcagccggaa atggctaaaa ctttgatcca gaacatctac tctatccgtg ctatcactca
     4921 gtggttccgt tctgctatcg aagctaaacg tttgaaagac gctgacccga aattgatgac
     4981 tgacgtatac gtatctttgt tccagggttt gttcttctgg ccgcaggtaa tgcacttgga
     5041 cttggaaccg cacggtgaag aattgtctca gaaaatcgaa actttgacta ctatcttctt
     5101 gcagtcttac ggtgtagctg aataacaatg aatgggcttt cctcggtacc aaattccaga
     5161 aaagaggcct cccgaaaggg gggccttttt tcgttttggt ccctaattgc ctatcttcca
     5221 gtgatggaac agcatttgtg cattggctgc aacaatcagc cttacttgtg cctgttctat
     5281 ttccgaaccg accgcttgta tgaatgcatc aaaattcgtt ttctctacgt tggattcctt
     5341 gttgctcata ttgtgatgat aatttctaca aatatagtca ttggtaacta tctatgaaac
     5401 tgtttgatac ttttatcagt ccagtagttt tacaaggtct ttcttcattt cttcgtcaat
     5461 atccctgtat cgtctgaaag ctttgcttcc ctccttgtgt cccgacagtg cggaaacaag
     5521 gttcgggtct ttcacttttt tatagatatt gccgataaac gtacgtcttg ccagatggct
     5581 gcttgccact tcataaatag gtcgtttgat ttcgttgtgc gtcaacgggt ctaagattgt
     5641 tacgatgcgg tcaactccag ctaatttgaa tatctttttt atggcatcat tgtacttttg
     5701 ctcggatatg aacggcaaca gttttccctc atattctttg tagcgttcaa ggatttcttt
     5761 cgctttgtcg ttaagtggaa cacgtaccgt aaccggattc ccctctttgg ttttcttggg
     5821 aatatattct atggcttcat tgaccacatt tagtttggtc attcggtaca ggtcgctcac
     5881 cctgcatcct atcagtgtct gaaatatgaa tatatccctc tgtattgcca gttgtggggt
     5941 ggcagaaagg tctgcattaa aaatcctgtc cctttcttcg agtgttatat aataaggtgt
     6001 accatatgta cactcctcta tcggaaactt gtcgaaaggt ctgtttgtgg tgcgtttgtt
     6061 atcgaagcac cacaggaaga atgtgcgtat tcttgaaaaa cagtctatca gcgtgttttt
     6121 gcttctgggc tgtggtgtcc tcttttcggg aatggcttca taaatgctcg ggtaaagttc
     6181 ataatactgg tattcgttct gaaagaaatc ccacatatcc cgaagcgtgt caggtgttac
     6241 caaatccaca tcaaggataa agcccttttg tcctctcttt gtagccctta catatagttc
     6301 ataacgcagt aaggctcttt tgacaacccg gaaatttttc tttcgtactt ccgacaaagg
     6361 gtgcttgttt agaaattcat cgaatagttc tccaatggta ggcttgataa ccacttcctc
     6421 cggcaaaaaa tatttttcag gatggtaaaa tttatcaagt gttgttttta gccattcttt
     6481 gtctattgct tccttttctt gttgatatac cttctcgata taggttttca actggcgtat
     6541 ctcttcattt atatgggtac gcatttcttc attgcagaca gctttcgttt ttacacattc
     6601 gtctttatca tcccatagat tgggattgat ggctaactga gtaggagcaa cggagtctaa
     6661 ctgccttccg tttctaaaac ggacatagat tgtagccatg gattctgtat catatcgttt
     6721 ggctgctttt ttaatgataa aggttacttt catagacttt caggttgaat tttactctgc
     6781 tgcaaatata aatattttcc ccagcatttt ccccacatct gctaaatatt ttgcaattcg
     6841 attaaacttg gattaaaatt taataggatt ataacatatt gaaatacagt gtaattgtgg
     6901 cgtttttccg catttttctt tttacccaca ttttccccac tttaattatt tcaaatgcgg
     6961 ggtctgggta caagaaagaa agctaagtat ttgatagttc aatacttagc tttttctttt
     7021 gcttgaattt tccccacatt ttccccacac gtgcaaaaaa tatagcagta agtcattatt
     7081 tcttttggtt gaacgtagag agtagcgata ttaaaagaat ccgat
//
