LOCUS       sequence-313939-        7139 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bile Acid inducible sgRNA-6 with PM6 driving Nanoluc expression (NOT
            Gate 6), pNBU1 backbone, AmpR, ErmR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7139)
  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 7139)
  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: pMT493
FEATURES             Location/Qualifiers
     source          1..7139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        99..174
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        364..413
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             join(5346..5414,5415..6206)
                     /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(5564..5583)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            6337..6446
                     /note="incP origin of transfer"
     rep_origin      6617..7005
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 atttaaatgt acacccgttt gtttacttta ctttagttat gtacctttgt cggcaatgta
       61 cacccgaaag tttactttca aagataaaat ggtttagtgt tttagagcta gaaatagcaa
      121 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt
      181 tggatcccca attattgaac accctaacgg gtgttttttt ttttttggtc tcccgcggcc
      241 gcgccgcggc cgctgctgtc ataggccggt tgtccggtag tccggatgat ttaaggaaga
      301 tttctcttct tttttcactt tctccgaaca acctccaact aaaccatttt atctttgcag
      361 tagctgtcac cggatgtgct ttccggtctg atgagtccgt gaggacgaaa cagcctctac
      421 aaataatttt gtttaacgca ttttaaaata aaataaatta tttatttaat taaacgaata
      481 tggtttttac tctggaagat tttgttggcg attggcgtca gaccgcgggt tataatttgg
      541 atcaagtcct ggaacagggt ggcgtaagct ctctgttcca gaacctgggt gtgagcgtga
      601 cgccgattca gcgcatcgtt ctgtccggcg agaacggtct gaaaattgat attcatgtga
      661 tcatcccgta cgaaggcctg agcggtgacc aaatgggtca aatcgagaaa atctttaaag
      721 tcgtctaccc agttgacgat caccacttca aggttatctt gcattacggt acgctggtga
      781 ttgatggtgt gaccccgaat atgattgact atttcggccg tccgtatgaa ggcattgccg
      841 tttttgacgg taaaaagatc accgtcaccg gtaccctgtg gaatggcaat aagattattg
      901 acgagcgtct gattaacccg gacggcagcc tgctgttccg cgtgaccatc aacggtgtca
      961 cgggttggcg tctgtgcgag cgcatcctgg cataatgaac tgggatccct cggtaccaaa
     1021 ttttcgaaaa aagacgctga aaagcgtctt ttttcgtttt ggtccgcggc cgccggccgc
     1081 tgatctggaa gaagcaatga aagctgctgt taagtctccg aatcaggtat tgttcctgac
     1141 aggtgtattc ccatccggta aacgcggata ctttgcagtt gatctgactc aggaataaat
     1201 tataaattaa ggtaagaaga ttgtaggata agctaatgaa atagaaaaag gatgccgtca
     1261 cacaacttgt cggcattctt ttttgtttta ttagttgaaa atatagtgaa aaagttgcct
     1321 aaatatgtat gttaacaaat tatttgtcgt aactttgcac tccaaatctg tttttaacat
     1381 atggcactaa tgcagaaaaa attgactcgt tctcagcaga aacacttgga catcatcaac
     1441 gctgctaaag aagaattcat cgaattcggt ttcttggctg ctaacatgga ccgtatcact
     1501 tcttctgctg aagtatctaa acgtactttg taccgtcact tcgaatctaa agaagtattg
     1561 ttcgaatctg tattgactat catcaacgac tctgtaaacg aatctatctc ttaccacttc
     1621 gacccgaaca aatctactga agaacagttg actgaaatcg cttacaaaga aatcgacgta
     1681 ttgtacaaaa cttacggtat cgctttggct cgtactatcg taatggaatt cttgcgtcag
     1741 ccggaaatgg ctaaaacttt gatccagaac atctactcta tccgtgctat cactcagtgg
     1801 ttccgttctg ctatcgaagc taaacgtttg aaagacgctg acccgaaatt gatgactgac
     1861 gtatacgtat ctttgttcca gggtttgttc ttctggccgc aggtaatgca cttggacttg
     1921 gaaccgcacg gtgaagaatt gtctcagaaa atcgaaactt tgactactat cttcttgcag
     1981 tcttacggtg tagctgaata acaatgaatg ggctttcctc ggtaccaaat tccagaaaag
     2041 aggcctcccg aaaggggggc cttttttcgt tttggtccct aattgcctat cttccagtga
     2101 tggaacagca tttgtgcatt ggctgcaaca atcagcctta cttgtgcctg ttctatttcc
     2161 gaaccgaccg cttgtatgaa tgcatcaaaa ttcgttttct ctacgttgga ttccttgttg
     2221 ctcatattgt gatgataatt tctacaaata tagtcattgg taactatcta tgaaactgtt
     2281 tgatactttt atcagtccag tagttttaca aggtctttct tcatttcttc gtcaatatcc
     2341 ctgtatcgtc tgaaagcttt gcttccctcc ttgtgtcccg acagtgcgga aacaaggttc
     2401 gggtctttca cttttttata gatattgccg ataaacgtac gtcttgccag atggctgctt
     2461 gccacttcat aaataggtcg tttgatttcg ttgtgcgtca acgggtctaa gattgttacg
     2521 atgcggtcaa ctccagctaa tttgaatatc ttttttatgg catcattgta cttttgctcg
     2581 gatatgaacg gcaacagttt tccctcatat tctttgtagc gttcaaggat ttctttcgct
     2641 ttgtcgttaa gtggaacacg taccgtaacc ggattcccct ctttggtttt cttgggaata
     2701 tattctatgg cttcattgac cacatttagt ttggtcattc ggtacaggtc gctcaccctg
     2761 catcctatca gtgtctgaaa tatgaatata tccctctgta ttgccagttg tggggtggca
     2821 gaaaggtctg cattaaaaat cctgtccctt tcttcgagtg ttatataata aggtgtacca
     2881 tatgtacact cctctatcgg aaacttgtcg aaaggtctgt ttgtggtgcg tttgttatcg
     2941 aagcaccaca ggaagaatgt gcgtattctt gaaaaacagt ctatcagcgt gtttttgctt
     3001 ctgggctgtg gtgtcctctt ttcgggaatg gcttcataaa tgctcgggta aagttcataa
     3061 tactggtatt cgttctgaaa gaaatcccac atatcccgaa gcgtgtcagg tgttaccaaa
     3121 tccacatcaa ggataaagcc cttttgtcct ctctttgtag cccttacata tagttcataa
     3181 cgcagtaagg ctcttttgac aacccggaaa tttttctttc gtacttccga caaagggtgc
     3241 ttgtttagaa attcatcgaa tagttctcca atggtaggct tgataaccac ttcctccggc
     3301 aaaaaatatt tttcaggatg gtaaaattta tcaagtgttg tttttagcca ttctttgtct
     3361 attgcttcct tttcttgttg atataccttc tcgatatagg ttttcaactg gcgtatctct
     3421 tcatttatat gggtacgcat ttcttcattg cagacagctt tcgtttttac acattcgtct
     3481 ttatcatccc atagattggg attgatggct aactgagtag gagcaacgga gtctaactgc
     3541 cttccgtttc taaaacggac atagattgta gccatggatt ctgtatcata tcgtttggct
     3601 gcttttttaa tgataaaggt tactttcata gactttcagg ttgaatttta ctctgctgca
     3661 aatataaata ttttccccag cattttcccc acatctgcta aatattttgc aattcgatta
     3721 aacttggatt aaaatttaat aggattataa catattgaaa tacagtgtaa ttgtggcgtt
     3781 tttccgcatt tttcttttta cccacatttt ccccacttta attatttcaa atgcggggtc
     3841 tgggtacaag aaagaaagct aagtatttga tagttcaata cttagctttt tcttttgctt
     3901 gaattttccc cacattttcc ccacacgtgc aaaaaatata gcagtaagtc attatttctt
     3961 ttggttgaac gtagagagta gcgatattaa aagaatccga tgagaaaaga ctaatattta
     4021 tctatccatt cagtttgatt tctcaggact ttacatcgtc ctgaaagtat ttgttggtac
     4081 cgaggacgcg taaacattta cagttgcatg tggcctattg tttttagccg ttaaatattt
     4141 tataactatt aaatagcgat acaaattgtt cgaaactaat attgtttata tcatatattc
     4201 tcgcatgttt taaagcttta ttaaattgat tttttgtaaa cagtttttcg tactctttgt
     4261 taacccattt cattacaaaa gtttcatatt tttttctctc tttaaatgcc atttttgctg
     4321 gctttctttt taatacaatt aatgtgctat ccactttagg ttttggatgg aaataatacc
     4381 taggaatttt tgctaatata gaaatatcta cctctgccat taacagcaat gctagtgatc
     4441 tgtttgtatc taataacatt ttagcaaaac catattccac tattaaataa cttattgtgg
     4501 ctgaactttc aaaaacaatt tttcgaatta tatttgtgct tatgttgtaa ggtatgctgc
     4561 caaatatttt atatggattg tggctaggaa atgtaaattt cagtatatca tcatttacta
     4621 tttgatagtt aggataattt aagagcttat tacgagttac ctcacataat ttagaatcaa
     4681 tttctatcgc cgttacaaaa ttacatctct ttaccaatcc agcagtaaaa tgacctttcc
     4741 ctgcacctat ttcaaagatg ttatcttttt catctaaact tatgcaattc attatttttt
     4801 ctatgtgata ttttgaagta ataaaatttt gactatcttt tatatttact ttgttcatta
     4861 taacctctcc ttaatttatt gcatctcttt tcgaatattt atgttttttg agaaaagaac
     4921 gtactcatgg ttcatcccga tatgcgtatc ggtctgtata tcagcaactt tctatgtgtt
     4981 tcaactacaa tagtcatcta ttctcatctt tctgagtcca ccccctgcaa agcccctctt
     5041 tacgacataa aaattcggtc ggaaaaggta tgcaaaagat gtttctctct ttaagagaaa
     5101 ctcttcggga tgcaaaaata tgaaaataac tccaattcac caaattatat agcgactttt
     5161 ttacaaaatg ctaaaatttg ttgatttccg tcaagcaatt gttgagcaaa aatgtctttt
     5221 acgataaaat gatacctcaa tatcaactgt ttagcaaaac gatatttctc ttaaagagag
     5281 aaacaccttt ttgttcacca atccccgact tttaatcccg cggccatgat tgaaaaagga
     5341 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5401 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5461 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5521 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5581 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5641 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5701 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5761 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5821 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5881 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5941 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6001 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6061 ggtcacgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6121 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6181 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcataa cgcgtcaatt
     6241 cgagggggat caattccgtg ataggtgggc tgcccttcct ggttggcttg gtttcatcag
     6301 ccatccgctt gccctcatct gttacgccgg cggtagccgg ccagcctcgc agagcaggat
     6361 tcccgttgag caccgccagg tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg
     6421 gtgggcctac ttcacctatc ctgcccggct gacgccgttg gatacaccaa ggaaagtcta
     6481 cacgaaccct ttggcaaaat cctgtatatc gtgcgaaaaa ggatggatat accgaaaaaa
     6541 tcgctataat gaccccgaag cagggttatg cagcggaaaa cggaattgat ccggccacga
     6601 tgcgtccggc gtagaggatc tgaagatcag cagttcaacc tgttgatagt acgtactaag
     6661 ctctcatgtt tcacgtacta agctctcatg tttaacgtac taagctctca tgtttaacga
     6721 actaaaccct catggctaac gtactaagct ctcatggcta acgtactaag ctctcatgtt
     6781 tcacgtacta agctctcatg tttgaacaat aaaattaata taaatcagca acttaaatag
     6841 cctctaaggt tttaagtttt ataagaaaaa aaagaatata taaggctttt aaagctttta
     6901 aggtttaacg gttgtggaca acaagccagg gatgtaacgc actgagaagc ccttagagcc
     6961 tctcaaagca attttgagtg acacaggaac acttaacggc tgacatggga attcccctcc
     7021 accgcggtgt attaatgcgg ctgcaatttt tttgggcggg gccgcccaaa aaaatcctag
     7081 caccctgcag ggtctcccgc tttacaaaga aaattcgaca aactgttatt tttctatct
//
