LOCUS       sequence-313941-        7130 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bile Acid inducible sgRNA-7 with PM7 driving Nanoluc expression (NOT
            Gate 7), pNBU1 backbone, AmpR, ErmR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7130)
  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 7130)
  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: pMT494
FEATURES             Location/Qualifiers
     source          1..7130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        100..175
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        356..405
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             join(5338..5406,5407..6198)
                     /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(5556..5575)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            6329..6438
                     /note="incP origin of transfer"
     rep_origin      6609..6997
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 tatttaaatg tacacccgtt tgtttacttt actttagtta tgtacctttg tcggcaatgt
       61 acacccgaaa gtttactttc aaagatatat aactatatag ttttagagct agaaatagca
      121 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
      181 ttgacgaaca ataaggcctc ccaaatcggg gggccttttt tattgataac aaaagccgcg
      241 gccgctgctg tcataggccg gttgtccggt agtccggatg atttaaggaa gatttctctt
      301 cttttttcac tttctccgaa caacctccat atatagttat atatctttgc agtagctgtc
      361 accggatgtg ctttccggtc tgatgagtcc gtgaggacga aacagcctct acaaataatt
      421 ttgtttaacg cattttaaaa taaaataaat tatttattta attaaacgaa tatggttttt
      481 actctggaag attttgttgg cgattggcgt cagaccgcgg gttataattt ggatcaagtc
      541 ctggaacagg gtggcgtaag ctctctgttc cagaacctgg gtgtgagcgt gacgccgatt
      601 cagcgcatcg ttctgtccgg cgagaacggt ctgaaaattg atattcatgt gatcatcccg
      661 tacgaaggcc tgagcggtga ccaaatgggt caaatcgaga aaatctttaa agtcgtctac
      721 ccagttgacg atcaccactt caaggttatc ttgcattacg gtacgctggt gattgatggt
      781 gtgaccccga atatgattga ctatttcggc cgtccgtatg aaggcattgc cgtttttgac
      841 ggtaaaaaga tcaccgtcac cggtaccctg tggaatggca ataagattat tgacgagcgt
      901 ctgattaacc cggacggcag cctgctgttc cgcgtgacca tcaacggtgt cacgggttgg
      961 cgtctgtgcg agcgcatcct ggcataatga actgggatcc ctcggtacca aattttcgaa
     1021 aaaagacgct gaaaagcgtc ttttttcgtt ttggtccgcg gccgccggcc gctgatctgg
     1081 aagaagcaat gaaagctgct gttaagtctc cgaatcaggt attgttcctg acaggtgtat
     1141 tcccatccgg taaacgcgga tactttgcag ttgatctgac tcaggaataa attataaatt
     1201 aaggtaagaa gattgtagga taagctaatg aaatagaaaa aggatgccgt cacacaactt
     1261 gtcggcattc ttttttgttt tattagttga aaatatagtg aaaaagttgc ctaaatatgt
     1321 atgttaacaa attatttgtc gtaactttgc actccaaatc tgtttttaac atatggcact
     1381 aatgcagaaa aaattgactc gttctcagca gaaacacttg gacatcatca acgctgctaa
     1441 agaagaattc atcgaattcg gtttcttggc tgctaacatg gaccgtatca cttcttctgc
     1501 tgaagtatct aaacgtactt tgtaccgtca cttcgaatct aaagaagtat tgttcgaatc
     1561 tgtattgact atcatcaacg actctgtaaa cgaatctatc tcttaccact tcgacccgaa
     1621 caaatctact gaagaacagt tgactgaaat cgcttacaaa gaaatcgacg tattgtacaa
     1681 aacttacggt atcgctttgg ctcgtactat cgtaatggaa ttcttgcgtc agccggaaat
     1741 ggctaaaact ttgatccaga acatctactc tatccgtgct atcactcagt ggttccgttc
     1801 tgctatcgaa gctaaacgtt tgaaagacgc tgacccgaaa ttgatgactg acgtatacgt
     1861 atctttgttc cagggtttgt tcttctggcc gcaggtaatg cacttggact tggaaccgca
     1921 cggtgaagaa ttgtctcaga aaatcgaaac tttgactact atcttcttgc agtcttacgg
     1981 tgtagctgaa taacaatgaa tgggctttcc tcggtaccaa attccagaaa agaggcctcc
     2041 cgaaaggggg gccttttttc gttttggtcc ctaattgcct atcttccagt gatggaacag
     2101 catttgtgca ttggctgcaa caatcagcct tacttgtgcc tgttctattt ccgaaccgac
     2161 cgcttgtatg aatgcatcaa aattcgtttt ctctacgttg gattccttgt tgctcatatt
     2221 gtgatgataa tttctacaaa tatagtcatt ggtaactatc tatgaaactg tttgatactt
     2281 ttatcagtcc agtagtttta caaggtcttt cttcatttct tcgtcaatat ccctgtatcg
     2341 tctgaaagct ttgcttccct ccttgtgtcc cgacagtgcg gaaacaaggt tcgggtcttt
     2401 cactttttta tagatattgc cgataaacgt acgtcttgcc agatggctgc ttgccacttc
     2461 ataaataggt cgtttgattt cgttgtgcgt caacgggtct aagattgtta cgatgcggtc
     2521 aactccagct aatttgaata tcttttttat ggcatcattg tacttttgct cggatatgaa
     2581 cggcaacagt tttccctcat attctttgta gcgttcaagg atttctttcg ctttgtcgtt
     2641 aagtggaaca cgtaccgtaa ccggattccc ctctttggtt ttcttgggaa tatattctat
     2701 ggcttcattg accacattta gtttggtcat tcggtacagg tcgctcaccc tgcatcctat
     2761 cagtgtctga aatatgaata tatccctctg tattgccagt tgtggggtgg cagaaaggtc
     2821 tgcattaaaa atcctgtccc tttcttcgag tgttatataa taaggtgtac catatgtaca
     2881 ctcctctatc ggaaacttgt cgaaaggtct gtttgtggtg cgtttgttat cgaagcacca
     2941 caggaagaat gtgcgtattc ttgaaaaaca gtctatcagc gtgtttttgc ttctgggctg
     3001 tggtgtcctc ttttcgggaa tggcttcata aatgctcggg taaagttcat aatactggta
     3061 ttcgttctga aagaaatccc acatatcccg aagcgtgtca ggtgttacca aatccacatc
     3121 aaggataaag cccttttgtc ctctctttgt agcccttaca tatagttcat aacgcagtaa
     3181 ggctcttttg acaacccgga aatttttctt tcgtacttcc gacaaagggt gcttgtttag
     3241 aaattcatcg aatagttctc caatggtagg cttgataacc acttcctccg gcaaaaaata
     3301 tttttcagga tggtaaaatt tatcaagtgt tgtttttagc cattctttgt ctattgcttc
     3361 cttttcttgt tgatatacct tctcgatata ggttttcaac tggcgtatct cttcatttat
     3421 atgggtacgc atttcttcat tgcagacagc tttcgttttt acacattcgt ctttatcatc
     3481 ccatagattg ggattgatgg ctaactgagt aggagcaacg gagtctaact gccttccgtt
     3541 tctaaaacgg acatagattg tagccatgga ttctgtatca tatcgtttgg ctgctttttt
     3601 aatgataaag gttactttca tagactttca ggttgaattt tactctgctg caaatataaa
     3661 tattttcccc agcattttcc ccacatctgc taaatatttt gcaattcgat taaacttgga
     3721 ttaaaattta ataggattat aacatattga aatacagtgt aattgtggcg tttttccgca
     3781 tttttctttt tacccacatt ttccccactt taattatttc aaatgcgggg tctgggtaca
     3841 agaaagaaag ctaagtattt gatagttcaa tacttagctt tttcttttgc ttgaattttc
     3901 cccacatttt ccccacacgt gcaaaaaata tagcagtaag tcattatttc ttttggttga
     3961 acgtagagag tagcgatatt aaaagaatcc gatgagaaaa gactaatatt tatctatcca
     4021 ttcagtttga tttctcagga ctttacatcg tcctgaaagt atttgttggt accgaggacg
     4081 cgtaaacatt tacagttgca tgtggcctat tgtttttagc cgttaaatat tttataacta
     4141 ttaaatagcg atacaaattg ttcgaaacta atattgttta tatcatatat tctcgcatgt
     4201 tttaaagctt tattaaattg attttttgta aacagttttt cgtactcttt gttaacccat
     4261 ttcattacaa aagtttcata tttttttctc tctttaaatg ccatttttgc tggctttctt
     4321 tttaatacaa ttaatgtgct atccacttta ggttttggat ggaaataata cctaggaatt
     4381 tttgctaata tagaaatatc tacctctgcc attaacagca atgctagtga tctgtttgta
     4441 tctaataaca ttttagcaaa accatattcc actattaaat aacttattgt ggctgaactt
     4501 tcaaaaacaa tttttcgaat tatatttgtg cttatgttgt aaggtatgct gccaaatatt
     4561 ttatatggat tgtggctagg aaatgtaaat ttcagtatat catcatttac tatttgatag
     4621 ttaggataat ttaagagctt attacgagtt acctcacata atttagaatc aatttctatc
     4681 gccgttacaa aattacatct ctttaccaat ccagcagtaa aatgaccttt ccctgcacct
     4741 atttcaaaga tgttatcttt ttcatctaaa cttatgcaat tcattatttt ttctatgtga
     4801 tattttgaag taataaaatt ttgactatct tttatattta ctttgttcat tataacctct
     4861 ccttaattta ttgcatctct tttcgaatat ttatgttttt tgagaaaaga acgtactcat
     4921 ggttcatccc gatatgcgta tcggtctgta tatcagcaac tttctatgtg tttcaactac
     4981 aatagtcatc tattctcatc tttctgagtc caccccctgc aaagcccctc tttacgacat
     5041 aaaaattcgg tcggaaaagg tatgcaaaag atgtttctct ctttaagaga aactcttcgg
     5101 gatgcaaaaa tatgaaaata actccaattc accaaattat atagcgactt ttttacaaaa
     5161 tgctaaaatt tgttgatttc cgtcaagcaa ttgttgagca aaaatgtctt ttacgataaa
     5221 atgatacctc aatatcaact gtttagcaaa acgatatttc tcttaaagag agaaacacct
     5281 ttttgttcac caatccccga cttttaatcc cgcggccatg attgaaaaag gaagagtatg
     5341 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5401 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5461 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5521 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5581 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5641 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5701 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5761 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5821 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5881 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5941 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6001 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtcacgc
     6061 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6121 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6181 ctgattaagc attggtaact gtcagaccaa gtttactcat aacgcgtcaa ttcgaggggg
     6241 atcaattccg tgataggtgg gctgcccttc ctggttggct tggtttcatc agccatccgc
     6301 ttgccctcat ctgttacgcc ggcggtagcc ggccagcctc gcagagcagg attcccgttg
     6361 agcaccgcca ggtgcgaata agggacagtg aagaaggaac acccgctcgc gggtgggcct
     6421 acttcaccta tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc tacacgaacc
     6481 ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa aatcgctata
     6541 atgaccccga agcagggtta tgcagcggaa aacggaattg atccggccac gatgcgtccg
     6601 gcgtagagga tctgaagatc agcagttcaa cctgttgata gtacgtacta agctctcatg
     6661 tttcacgtac taagctctca tgtttaacgt actaagctct catgtttaac gaactaaacc
     6721 ctcatggcta acgtactaag ctctcatggc taacgtacta agctctcatg tttcacgtac
     6781 taagctctca tgtttgaaca ataaaattaa tataaatcag caacttaaat agcctctaag
     6841 gttttaagtt ttataagaaa aaaaagaata tataaggctt ttaaagcttt taaggtttaa
     6901 cggttgtgga caacaagcca gggatgtaac gcactgagaa gcccttagag cctctcaaag
     6961 caattttgag tgacacagga acacttaacg gctgacatgg gaattcccct ccaccgcggt
     7021 gtattaatgc ggctgcaatt tttttgggcg gggccgccca aaaaaatcct agcaccctgc
     7081 agggtctccc gctttacaaa gaaaattcga caaactgtta tttttctatc
//
