LOCUS       sequence-313935-        9591 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  XOR circuit (A000 and P000 design) with aTC and Bile Acid as inputs,
            AmpR, ErmR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9591)
  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 9591)
  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: pMT462
FEATURES             Location/Qualifiers
     source          1..9591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        141..216
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    420..438
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    482..500
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        525..600
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        815..890
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        1134..1209
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        1424..1499
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    1660..1678
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1722..1740
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        1765..1840
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        2028..2077
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     misc_RNA        2858..2907
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             join(7844..7912,7913..8704)
                     /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(8062..8081)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            8835..8944
                     /note="incP origin of transfer"
     rep_origin      9115..9503
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 ctcccgctgg agtgctgtca taggccggtt gtccggtagt ccggatgatt taaggaagat
       61 ttctcttctt ttttcacttt ctccgaacaa cctccactta agttctatta tctttgcagt
      121 caaagataaa tcagagaaaa gttttagagc tagaaatagc aagttaaaat aaggctagtc
      181 cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tttggatcct tcagtcaaaa
      241 aacttaagac cgccggtctt gtccactacc ttgcagtaat gcggtggaca ggatcggcgg
      301 ttttcttttc tcttctcaag cggccgctac gttttgcacc cgctttccaa gagaagaaag
      361 ccttgttaaa ttgacttagt gtaaaagcgc agtactgctt gaccataaga acaaaaaaat
      421 ctctatcact gatagggata aagtttggaa gataaagcta aaagttctta tctttgcagt
      481 ctccctatca gtgatagaga cgcacaaaga tacttatgtt agaagtttta gagctagaaa
      541 tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc
      601 tttttttgga tccaacgcat gagaaagccc ccggaagatc accttccggg ggctttttta
      661 ttgcgcgcgg ccgcaggttt acaaagaaaa ttcgacaaac tgttattttt ctatctattt
      721 aaatgtacac ccgtttgttt actttacttt agttatgtac ctttgtcggc aatgtacacc
      781 cgaaagttta ctttcaaaga taaatcagag aaaagtttta gagctagaaa tagcaagtta
      841 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc tttttttgga
      901 tccttcagtc aaaaaactta agaccgccgg tcttgtccac taccttgcag taatgcggtg
      961 gacaggatcg gcggttttct tttctcttct caagcggccg cgctttgctg tcataggccg
     1021 gttgtccggt agtccggatg atttaaggaa gatttctctt cttttttcac tttctccgaa
     1081 caacctccac ttaagttcta ttatctttgc agtcaaagat acttatgtta gaagttttag
     1141 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
     1201 agtcggtgct ttttttggat ccaacgcatg agaaagcccc cggaagatca ccttccgggg
     1261 gcttttttat tgcgcgcggc cgccgcttta caaagaaaat tcgacaaact gttatttttc
     1321 tatctattta aatgtacacc cgtttgttta ctttacttta gttatgtacc tttgtcggca
     1381 atgtacaccc gaaagtttac tttcaaagat aatagaactt aaggttttag agctagaaat
     1441 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
     1501 ttttttggat ccaaaaaaaa aaaacaccct aacgggtgtt tttttttttt tggtctgccg
     1561 cggccgctgt cttttgcacc cgctttccaa gagaagaaag ccttgttaaa ttgacttagt
     1621 gtaaaagcgc agtactgctt gaccataaga acaaaaaaat ctctatcact gatagggata
     1681 aagtttggaa gataaagcta aaagttctta tctttgcagt ctccctatca gtgatagaga
     1741 cgcacaaaga taatagaact taaggtttta gagctagaaa tagcaagtta aaataaggct
     1801 agtccgttat caacttgaaa aagtggcacc gagtcggtgc tttttttgga tccaaaaaaa
     1861 aaaaacaccc taacgggtgt tttttttttt tttggtctgc cgcggccgca atgggagtgc
     1921 tgtcataggc cggttgtccg gtagtccgga tgatttaagg aagatttctc ttcttttttc
     1981 actttctccg aacaacctcc attttctctg atttatcttt gcagtagctg tcaccggatg
     2041 tgctttccgg tctgatgagt ccgtgaggac gaaacagcct ctacaaataa ttttgtttaa
     2101 cgcattttaa aataaaataa attatttatt taattaaacg aatatggttt ttactctgga
     2161 agattttgtt ggcgattggc gtcagaccgc gggttataat ttggatcaag tcctggaaca
     2221 gggtggcgta agctctctgt tccagaacct gggtgtgagc gtgacgccga ttcagcgcat
     2281 cgttctgtcc ggcgagaacg gtctgaaaat tgatattcat gtgatcatcc cgtacgaagg
     2341 cctgagcggt gaccaaatgg gtcaaatcga gaaaatcttt aaagtcgtct acccagttga
     2401 cgatcaccac ttcaaggtta tcttgcatta cggtacgctg gtgattgatg gtgtgacccc
     2461 gaatatgatt gactatttcg gccgtccgta tgaaggcatt gccgtttttg acggtaaaaa
     2521 gatcaccgtc accggtaccc tgtggaatgg caataagatt attgacgagc gtctgattaa
     2581 cccggacggc agcctgctgt tccgcgtgac catcaacggt gtcacgggtt ggcgtctgtg
     2641 cgagcgcatc ctggcataat gaactgggat ccctcggtac caaattttcg aaaaaagacg
     2701 ctgaaaagcg tcttttttcg ttttggtccg cggccgccgc cgctacgtgc tgtcataggc
     2761 cggttgtccg gtagtccgga tgatttaagg aagatttctc ttcttttttc actttctccg
     2821 aacaacctcc attctaacat aagtatcttt gcagtagctg tcaccggatg tgctttccgg
     2881 tctgatgagt ccgtgaggac gaaacagcct ctacaaataa ttttgtttaa cgcattttaa
     2941 aataaaataa attatttatt taattaaacg aatatggttt ttactctgga agattttgtt
     3001 ggcgattggc gtcagaccgc gggttataat ttggatcaag tcctggaaca gggtggcgta
     3061 agctctctgt tccagaacct gggtgtgagc gtgacgccga ttcagcgcat cgttctgtcc
     3121 ggcgagaacg gtctgaaaat tgatattcat gtgatcatcc cgtacgaagg cctgagcggt
     3181 gaccaaatgg gtcaaatcga gaaaatcttt aaagtcgtct acccagttga cgatcaccac
     3241 ttcaaggtta tcttgcatta cggtacgctg gtgattgatg gtgtgacccc gaatatgatt
     3301 gactatttcg gccgtccgta tgaaggcatt gccgtttttg acggtaaaaa gatcaccgtc
     3361 accggtaccc tgtggaatgg caataagatt attgacgagc gtctgattaa cccggacggc
     3421 agcctgctgt tccgcgtgac catcaacggt gtcacgggtt ggcgtctgtg cgagcgcatc
     3481 ctggcataat gaactgggat ccctcggtac caaattttcg aaaaaagacg ctgaaaagcg
     3541 tcttttttcg ttttggtccg cggccgccgg ccgcaatgtg atctggaaga agcaatgaaa
     3601 gctgctgtta agtctccgaa tcaggtattg ttcctgacag gtgtattccc atccggtaaa
     3661 cgcggatact ttgcagttga tctgactcag gaataaatta taaattaagg taagaagatt
     3721 gtaggataag ctaatgaaat agaaaaagga tgccgtcaca caacttgtcg gcattctttt
     3781 ttgttttatt agttgaaaat atagtgaaaa agttgcctaa atatgtatgt taacaaatta
     3841 tttgtcgtaa ctttgcactc caaatctgtt tttaacatat ggcactaatg cagaaaaaat
     3901 tgactcgttc tcagcagaaa cacttggaca tcatcaacgc tgctaaagaa gaattcatcg
     3961 aattcggttt cttggctgct aacatggacc gtatcacttc ttctgctgaa gtatctaaac
     4021 gtactttgta ccgtcacttc gaatctaaag aagtattgtt cgaatctgta ttgactatca
     4081 tcaacgactc tgtaaacgaa tctatctctt accacttcga cccgaacaaa tctactgaag
     4141 aacagttgac tgaaatcgct tacaaagaaa tcgacgtatt gtacaaaact tacggtatcg
     4201 ctttggctcg tactatcgta atggaattct tgcgtcagcc ggaaatggct aaaactttga
     4261 tccagaacat ctactctatc cgtgctatca ctcagtggtt ccgttctgct atcgaagcta
     4321 aacgtttgaa agacgctgac ccgaaattga tgactgacgt atacgtatct ttgttccagg
     4381 gtttgttctt ctggccgcag gtaatgcact tggacttgga accgcacggt gaagaattgt
     4441 ctcagaaaat cgaaactttg actactatct tcttgcagtc ttacggtgta gctgaataac
     4501 aatgaatggg ctttcctcgg taccaaattc cagaaaagag gcctcccgaa aggggggcct
     4561 tttttcgttt tggtccctaa ttgcctatct tccagtgatg gaacagcatt tgtgcattgg
     4621 ctgcaacaat cagccttact tgtgcctgtt ctatttccga accgaccgct tgtatgaatg
     4681 catcaaaatt cgttttctct acgttggatt ccttgttgct catattgtga tgataatttc
     4741 tacaaatata gtcattggta actatctatg aaactgtttg atacttttat cagtccagta
     4801 gttttacaag gtctttcttc atttcttcgt caatatccct gtatcgtctg aaagctttgc
     4861 ttccctcctt gtgtcccgac agtgcggaaa caaggttcgg gtctttcact tttttataga
     4921 tattgccgat aaacgtacgt cttgccagat ggctgcttgc cacttcataa ataggtcgtt
     4981 tgatttcgtt gtgcgtcaac gggtctaaga ttgttacgat gcggtcaact ccagctaatt
     5041 tgaatatctt ttttatggca tcattgtact tttgctcgga tatgaacggc aacagttttc
     5101 cctcatattc tttgtagcgt tcaaggattt ctttcgcttt gtcgttaagt ggaacacgta
     5161 ccgtaaccgg attcccctct ttggttttct tgggaatata ttctatggct tcattgacca
     5221 catttagttt ggtcattcgg tacaggtcgc tcaccctgca tcctatcagt gtctgaaata
     5281 tgaatatatc cctctgtatt gccagttgtg gggtggcaga aaggtctgca ttaaaaatcc
     5341 tgtccctttc ttcgagtgtt atataataag gtgtaccata tgtacactcc tctatcggaa
     5401 acttgtcgaa aggtctgttt gtggtgcgtt tgttatcgaa gcaccacagg aagaatgtgc
     5461 gtattcttga aaaacagtct atcagcgtgt ttttgcttct gggctgtggt gtcctctttt
     5521 cgggaatggc ttcataaatg ctcgggtaaa gttcataata ctggtattcg ttctgaaaga
     5581 aatcccacat atcccgaagc gtgtcaggtg ttaccaaatc cacatcaagg ataaagccct
     5641 tttgtcctct ctttgtagcc cttacatata gttcataacg cagtaaggct cttttgacaa
     5701 cccggaaatt tttctttcgt acttccgaca aagggtgctt gtttagaaat tcatcgaata
     5761 gttctccaat ggtaggcttg ataaccactt cctccggcaa aaaatatttt tcaggatggt
     5821 aaaatttatc aagtgttgtt tttagccatt ctttgtctat tgcttccttt tcttgttgat
     5881 ataccttctc gatataggtt ttcaactggc gtatctcttc atttatatgg gtacgcattt
     5941 cttcattgca gacagctttc gtttttacac attcgtcttt atcatcccat agattgggat
     6001 tgatggctaa ctgagtagga gcaacggagt ctaactgcct tccgtttcta aaacggacat
     6061 agattgtagc catggattct gtatcatatc gtttggctgc ttttttaatg ataaaggtta
     6121 ctttcataga ctttcaggtt gaattttact ctgctgcaaa tataaatatt ttccccagca
     6181 ttttccccac atctgctaaa tattttgcaa ttcgattaaa cttggattaa aatttaatag
     6241 gattataaca tattgaaata cagtgtaatt gtggcgtttt tccgcatttt tctttttacc
     6301 cacattttcc ccactttaat tatttcaaat gcggggtctg ggtacaagaa agaaagctaa
     6361 gtatttgata gttcaatact tagctttttc ttttgcttga attttcccca cattttcccc
     6421 acacgtgcaa aaaatatagc agtaagtcat tatttctttt ggttgaacgt agagagtagc
     6481 gatattaaaa gaatccgatg agaaaagact aatatttatc tatccattca gtttgatttc
     6541 tcaggacttt acatcgtcct gaaagtattt gttggtaccg aggacgcgta aacatttaca
     6601 gttgcatgtg gcctattgtt tttagccgtt aaatatttta taactattaa atagcgatac
     6661 aaattgttcg aaactaatat tgtttatatc atatattctc gcatgtttta aagctttatt
     6721 aaattgattt tttgtaaaca gtttttcgta ctctttgtta acccatttca ttacaaaagt
     6781 ttcatatttt tttctctctt taaatgccat ttttgctggc tttcttttta atacaattaa
     6841 tgtgctatcc actttaggtt ttggatggaa ataataccta ggaatttttg ctaatataga
     6901 aatatctacc tctgccatta acagcaatgc tagtgatctg tttgtatcta ataacatttt
     6961 agcaaaacca tattccacta ttaaataact tattgtggct gaactttcaa aaacaatttt
     7021 tcgaattata tttgtgctta tgttgtaagg tatgctgcca aatattttat atggattgtg
     7081 gctaggaaat gtaaatttca gtatatcatc atttactatt tgatagttag gataatttaa
     7141 gagcttatta cgagttacct cacataattt agaatcaatt tctatcgccg ttacaaaatt
     7201 acatctcttt accaatccag cagtaaaatg acctttccct gcacctattt caaagatgtt
     7261 atctttttca tctaaactta tgcaattcat tattttttct atgtgatatt ttgaagtaat
     7321 aaaattttga ctatctttta tatttacttt gttcattata acctctcctt aatttattgc
     7381 atctcttttc gaatatttat gttttttgag aaaagaacgt actcatggtt catcccgata
     7441 tgcgtatcgg tctgtatatc agcaactttc tatgtgtttc aactacaata gtcatctatt
     7501 ctcatctttc tgagtccacc ccctgcaaag cccctcttta cgacataaaa attcggtcgg
     7561 aaaaggtatg caaaagatgt ttctctcttt aagagaaact cttcgggatg caaaaatatg
     7621 aaaataactc caattcacca aattatatag cgactttttt acaaaatgct aaaatttgtt
     7681 gatttccgtc aagcaattgt tgagcaaaaa tgtcttttac gataaaatga tacctcaata
     7741 tcaactgttt agcaaaacga tatttctctt aaagagagaa acaccttttt gttcaccaat
     7801 ccccgacttt taatcccgcg gccatgattg aaaaaggaag agtatgagta ttcaacattt
     7861 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     7921 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     7981 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     8041 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     8101 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     8161 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     8221 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     8281 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     8341 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     8401 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     8461 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     8521 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tcacgcggta tcattgcagc
     8581 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     8641 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     8701 gtaactgtca gaccaagttt actcataacg cgtcaattcg agggggatca attccgtgat
     8761 aggtgggctg cccttcctgg ttggcttggt ttcatcagcc atccgcttgc cctcatctgt
     8821 tacgccggcg gtagccggcc agcctcgcag agcaggattc ccgttgagca ccgccaggtg
     8881 cgaataaggg acagtgaaga aggaacaccc gctcgcgggt gggcctactt cacctatcct
     8941 gcccggctga cgccgttgga tacaccaagg aaagtctaca cgaacccttt ggcaaaatcc
     9001 tgtatatcgt gcgaaaaagg atggatatac cgaaaaaatc gctataatga ccccgaagca
     9061 gggttatgca gcggaaaacg gaattgatcc ggccacgatg cgtccggcgt agaggatctg
     9121 aagatcagca gttcaacctg ttgatagtac gtactaagct ctcatgtttc acgtactaag
     9181 ctctcatgtt taacgtacta agctctcatg tttaacgaac taaaccctca tggctaacgt
     9241 actaagctct catggctaac gtactaagct ctcatgtttc acgtactaag ctctcatgtt
     9301 tgaacaataa aattaatata aatcagcaac ttaaatagcc tctaaggttt taagttttat
     9361 aagaaaaaaa agaatatata aggcttttaa agcttttaag gtttaacggt tgtggacaac
     9421 aagccaggga tgtaacgcac tgagaagccc ttagagcctc tcaaagcaat tttgagtgac
     9481 acaggaacac ttaacggctg acatgggaat tcccctccac cgcggtgtat taatgcggct
     9541 gcaatttttt tgggcggggc cgcccaaaaa aatcctagca ccctgcaggg t
//
