LOCUS       sequence-269613-        9657 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 9657)
  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 9657)
  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..9657
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1378..1446,1447..2238)
                     /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(1596..1615)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            2369..2478
                     /note="incP origin of transfer"
     rep_origin      2649..3037
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     misc_RNA        3266..3341
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    3545..3563
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3607..3625
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        3650..3725
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        3940..4015
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        4259..4334
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        4549..4624
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    4785..4803
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4847..4865
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        4890..4965
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        5152..5201
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     misc_RNA        5983..6032
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
ORIGIN
        1 agagtagcga tattaaaaga atccgatgag aaaagactaa tatttatcta tccattcagt
       61 ttgatttttc aggactttac atcgtcctga aagtatttgt tggtaccggt accgaggacg
      121 cgtaaacatt tacagttgca tgtggcctat tgtttttagc cgttaaatat tttataacta
      181 ttaaatagcg atacaaattg ttcgaaacta atattgttta tatcatatat tctcgcatgt
      241 tttaaagctt tattaaattg attttttgta aacagttttt cgtactcttt gttaacccat
      301 ttcattacaa aagtttcata tttttttctc tctttaaatg ccatttttgc tggctttctt
      361 tttaatacaa ttaatgtgct atccacttta ggttttggat ggaaataata cctaggaatt
      421 tttgctaata tagaaatatc tacctctgcc attaacagca atgctagtga tctgtttgta
      481 tctaataaca ttttagcaaa accatattcc actattaaat aacttattgt ggctgaactt
      541 tcaaaaacaa tttttcgaat tatatttgtg cttatgttgt aaggtatgct gccaaatatt
      601 ttatatggat tgtggctagg aaatgtaaat ttcagtatat catcatttac tatttgatag
      661 ttaggataat ttaagagctt attacgagtt acctcacata atttagaatc aatttctatc
      721 gccgttacaa aattacatct ctttaccaat ccagcagtaa aatgaccttt ccctgcacct
      781 atttcaaaga tgttatcttt ttcatctaaa cttatgcaat tcattatttt ttctatgtga
      841 tattttgaag taataaaatt ttgactatct tttatattta ctttgttcat tataacctct
      901 ccttaattta ttgcatctct tttcgaatat ttatgttttt tgagaaaaga acgtactcat
      961 ggttcatccc gatatgcgta tcggtctgta tatcagcaac tttctatgtg tttcaactac
     1021 aatagtcatc tattctcatc tttctgagtc caccccctgc aaagcccctc tttacgacat
     1081 aaaaattcgg tcggaaaagg tatgcaaaag atgtttctct ctttaagaga aactcttcgg
     1141 gatgcaaaaa tatgaaaata actccaattc accaaattat atagcgactt ttttacaaaa
     1201 tgctaaaatt tgttgatttc cgtcaagcaa ttgttgagca aaaatgtctt ttacgataaa
     1261 atgatacctc aatatcaact gtttagcaaa acgatatttc tcttaaagag agaaacacct
     1321 ttttgttcac caatccccga cttttaatcc cgcggccatg attgaaaaag gaagagtatg
     1381 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     1441 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     1501 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     1561 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     1621 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     1681 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     1741 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     1801 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     1861 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     1921 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     1981 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     2041 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtcacgc
     2101 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     2161 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     2221 ctgattaagc attggtaact gtcagaccaa gtttactcat aacgcgtcaa ttcgaggggg
     2281 atcaattccg tgataggtgg gctgcccttc ctggttggct tggtttcatc agccatccgc
     2341 ttgccctcat ctgttacgcc ggcggtagcc ggccagcctc gcagagcagg attcccgttg
     2401 agcaccgcca ggtgcgaata agggacagtg aagaaggaac acccgctcgc gggtgggcct
     2461 acttcaccta tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc tacacgaacc
     2521 ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa aatcgctata
     2581 atgaccccga agcagggtta tgcagcggaa aacggaattg atccggccac gatgcgtccg
     2641 gcgtagagga tctgaagatc agcagttcaa cctgttgata gtacgtacta agctctcatg
     2701 tttcacgtac taagctctca tgtttaacgt actaagctct catgtttaac gaactaaacc
     2761 ctcatggcta acgtactaag ctctcatggc taacgtacta agctctcatg tttcacgtac
     2821 taagctctca tgtttgaaca ataaaattaa tataaatcag caacttaaat agcctctaag
     2881 gttttaagtt ttataagaaa aaaaagaata tataaggctt ttaaagcttt taaggtttaa
     2941 cggttgtgga caacaagcca gggatgtaac gcactgagaa gcccttagag cctctcaaag
     3001 caattttgag tgacacagga acacttaacg gctgacatgg gaattcccct ccaccgcggt
     3061 ggattaatgc ggctgcaatt tttttgggcg gggccgccca aaaaaatcct agcaccctgc
     3121 agggtctccc gctggagtgc tgtcataggc cggttgtccg gtagtccgga tgatttaagg
     3181 aagatttctc ttcttttttc actttctccg aacaacctcc acttaagttc tattatcttt
     3241 gcagtcaaag ataaatcaga gaaaagtttt agagctagaa atagcaagtt aaaataaggc
     3301 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg ctttttttgg atccttcagc
     3361 caaaaaactt aagaccgccg gtcttgtcca ctaccttgca gtaatgcggt ggacaggatc
     3421 ggcggttttc ttttctcttc tcaagcggcc gctacgtttt gcacccgctt tccaagagaa
     3481 gaaagccttg ttaaattgac ttagtgtaaa agcgcagtac tgcttgacca taagaacaaa
     3541 aaaatctcta tcactgatag ggataaagtt tggaagataa agctaaaagt tcttatcttt
     3601 gcagtctccc tatcagtgat agagacgcac aaagatactt atgttagaag ttttagagct
     3661 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
     3721 ggtgcttttt ttggatccaa cgcatgagaa agcccccgga agatcacctt ccgggggctt
     3781 ttttattgcg cgcggccgca ggtttacaaa gaaaattcga caaactgtta tttttctatc
     3841 tatttaaatg tacacccgtt tgtttacttt actttagtta tgtacctttg tcggcaatgt
     3901 acacccgaaa gtttactttc aaagataaat cagagaaaag ttttagagct agaaatagca
     3961 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
     4021 ttggatcctt cagccaaaaa acttaagacc gccggtcttg tccactacct tgcagtaatg
     4081 cggtggacag gatcggcggt tttcttttct cttctcaagc ggccgcgctt tgctgtcata
     4141 ggccggttgt ccggtagtcc ggatgattta aggaagattt ctcttctttt ttcactttct
     4201 ccgaacaacc tccacttaag ttctattatc tttgcagtca aagatactta tgttagaagt
     4261 tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg
     4321 caccgagtcg gtgctttttt tggatccaac gcatgagaaa gcccccggaa gatcaccttc
     4381 cgggggcttt tttattgcgc gcggccgccg ctttacaaag aaaattcgac aaactgttat
     4441 ttttctatct atttaaatgt acacccgttt gtttacttta ctttagttat gtacctttgt
     4501 cggcaatgta cacccgaaag tttactttca aagataatag aacttaaggt tttagagcta
     4561 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     4621 gtgctttttt tggatccaaa aaaaaaaaac accctaacgg gtgttttttt ttttttggtc
     4681 tgccgcggcc gctgtctttt gcacccgctt tccaagagaa gaaagccttg ttaaattgac
     4741 ttagtgtaaa agcgcagtac tgcttgacca taagaacaaa aaaatctcta tcactgatag
     4801 ggataaagtt tggaagataa agctaaaagt tcttatcttt gcagtctccc tatcagtgat
     4861 agagacgcac aaagataata gaacttaagg ttttagagct agaaatagca agttaaaata
     4921 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttggatccaa
     4981 aaaaaaaaaa caccctaacg ggtgtttttt tttttttggt ctgccgcggc cgcaatggga
     5041 gtgctgtcat aggccggttg tccggtagtc cggatgattt aaggaagatt tctcttcttt
     5101 tttcactttc tccgaacaac ctccattttc tctgatttat ctttgcagta gctgtcaccg
     5161 gatgtgcttt ccggtctgat gagtccgtga ggacgaaaca gcctctacaa ataattttgt
     5221 ttaacgcatt ttaaaataaa ataaattatt tatttaatta aacgaatatg gtttttactc
     5281 tggaagattt tgttggcgat tggcgtcaga ccgcgggtta taatttggat caagtcctgg
     5341 aacagggtgg cgtaagctct ctgttccaga acctgggtgt gagcgtgacg ccgattcagc
     5401 gcatcgttct gtccggcgag aacggtctga aaattgatat tcatgtgatc atcccgtacg
     5461 aaggcctgag cggtgaccaa atgggtcaaa tcgagaaaat ctttaaagtc gtctacccag
     5521 ttgacgatca ccacttcaag gttatcttgc attacggtac gctggtgatt gatggtgtga
     5581 ccccgaatat gattgactat ttcggccgtc cgtatgaagg cattgccgtt tttgacggta
     5641 aaaagatcac cgtcaccggt accctgtgga atggcaataa gattattgac gagcgtctga
     5701 ttaacccgga cggcagcctg ctgttccgcg tgaccatcaa cggtgtcacg ggttggcgtc
     5761 tgtgcgagcg catcctggca taatgaactg ggatccctcg gtaccaaatt ttcgaaaaaa
     5821 gacgctgaaa agcgtctttt ttcgttttgg tccgcggccg ccggccgcta cgtgctgtca
     5881 taggccggtt gtccggtagt ccggatgatt taaggaagat ttctcttctt ttttcacttt
     5941 ctccgaacaa cctccattct aacataagta tctttgcagt agctgtcacc ggatgtgctt
     6001 tccggtctga tgagtccgtg aggacgaaac agcctctaca aataattttg tttaacgcat
     6061 tttaaaataa aataaattat ttatttaatt aaacgaatat ggtttttact ctggaagatt
     6121 ttgttggcga ttggcgtcag accgcgggtt ataatttgga tcaagtcctg gaacagggtg
     6181 gcgtaagctc tctgttccag aacctgggtg tgagcgtgac gccgattcag cgcatcgttc
     6241 tgtccggcga gaacggtctg aaaattgata ttcatgtgat catcccgtac gaaggcctga
     6301 gcggtgacca aatgggtcaa atcgagaaaa tctttaaagt cgtctaccca gttgacgatc
     6361 accacttcaa ggttatcttg cattacggta cgctggtgat tgatggtgtg accccgaata
     6421 tgattgacta tttcggccgt ccgtatgaag gcattgccgt ttttgacggt aaaaagatca
     6481 ccgtcaccgg taccctgtgg aatggcaata agattattga cgagcgtctg attaacccgg
     6541 acggcagcct gctgttccgc gtgaccatca acggtgtcac gggttggcgt ctgtgcgagc
     6601 gcatcctggc ataatgaact gggatccctc ggtaccaaat tttcgaaaaa agacgctgaa
     6661 aagcgtcttt tttcgttttg gtccgcggcc gccggccgca atgtgatctg gaagaagcaa
     6721 tgaaagctgc tgttaagtct ccgaatcagg tattgttcct gacaggtgta ttcccatccg
     6781 gtaaacgcgg atactttgca gttgatctga ctcaggaata aattataaat taaggtaaga
     6841 agattgtagg ataagctaat gaaatagaaa aaggatgccg tcacacaact tgtcggcatt
     6901 cttttttgtt ttattagttg aaaatatagt gaaaaagttg cctaaatatg tatgttaaca
     6961 aattatttgt cgtaactttg cactccaaat ctgtttttaa catatggcac taatgcagaa
     7021 aaaattgact cgttctcagc agaaacactt ggacatcatc aacgctgcta aagaagaatt
     7081 catcgaattc ggtttcttgg ctgctaacat ggaccgtatc acttcttctg ctgaagtatc
     7141 taaacgtact ttgtaccgtc acttcgaatc taaagaagta ttgttcgaat ctgtattgac
     7201 tatcatcaac gactctgtaa acgaatctat ctcttaccac ttcgacccga acaaatctac
     7261 tgaagaacag ttgactgaaa tcgcttacaa agaaatcgac gtattgtaca aaacttacgg
     7321 tatcgctttg gctcgtacta tcgtaatgga attcttgcgt cagccggaaa tggctaaaac
     7381 tttgatccag aacatctact ctatccgtgc tatcactcag tggttccgtt ctgctatcga
     7441 agctaaacgt ttgaaagacg ctgacccgaa attgatgact gacgtatacg tatctttgtt
     7501 ccagggtttg ttcttctggc cgcaggtaat gcacttggac ttggaaccgc acggtgaaga
     7561 attgtctcag aaaatcgaaa ctttgactac tatcttcttg cagtcttacg gtgtagctga
     7621 ataacaatga atgggctttc ctcggtacca aattccagaa aagaggcctc ccgaaagggg
     7681 ggcctttttt cgttttggtc cctaattgcc tatcttccag tgatggaaca gcatttgtgc
     7741 attggctgca acaatcagcc ttacttgtgc ctgttctatt tccgaaccga ccgcttgtat
     7801 gaatccatca aaattcgttt tctctatgtt ggattccttg ttgctcatat tgtgatgata
     7861 atttctacaa atatagtcat tggtaactat ctatgaaact gtttgatact tttactcatc
     7921 ggattctttt aatatcgcta ctctctacgt tcaaccaaaa gaaataatga cttactgcta
     7981 tattttttgc acgtgtgggg aaaatgtggg gaaaattcaa gcaaaagaaa aagctaagta
     8041 ttgaactatc aaatacttag ctttctttct tgtacccaga ccccgcattt gaaataatta
     8101 aagtggggaa aatgtgggta aaaagaaaaa tgcggaaaaa cgccacaatt acactgtatt
     8161 tcaatatgtt ataatcctat taaattttaa tccaagttta atcgaattgc aaaatattta
     8221 gcagatgtgg ggaaaatgct ggggaaaata tttatatttg cagcagagta aaattcaacc
     8281 tgaaagtcta tgaaagtaac ctttatcatt aaaaaagcag ccaaacgata tgatacagaa
     8341 tccatggcta caatctatgt ccgttttaga aacggaaggc agttagactc cgttgctcct
     8401 actcagttag ccatcaatcc caatctatgg gatgataaag acgaatgtgt aaaaacgaaa
     8461 gctgtctgca atgaagaaat gcgtacccat ataaatgaag agatacgcca gttgaaaacc
     8521 tatatcgaga aggtatatca acaagaaaag gaagcaatag acaaagaatg gctaaaaaca
     8581 acacttgata aattttacca tcctgaaaaa tattttttgc cggaggaagt ggttatcaag
     8641 cctaccattg gagaactatt cgatgaattt ctaaacaagc accctttgtc ggaagtacga
     8701 aagaaaaatt tccgggttgt caaaagagcc ttactgcgtt atgaactata tgtaagggct
     8761 acaaagagag gacaaaaggg ctttatcctt gatgtggatt tggtaacacc tgacacgctt
     8821 cgggatatgt gggatttctt tcagaacgaa taccagtatt atgaacttta cccgagcatt
     8881 tatgaagcca ttcccgaaaa gaggacacca cagcccagaa gcaaaaacac gctgatagac
     8941 tgtttttcaa gaatacgcac attcttcctg tggtgcttcg ataacaaacg caccacaaac
     9001 agacctttcg acaagtttcc gatagaggag tgtacatatg gtacacctta ttatataaca
     9061 ctcgaagaaa gggacaggat ttttaatgca gacctttctg ccaccccaca actggcaata
     9121 cagagggata tattcatatt tcagacactg ataggatgca gggtgagcga cctgtaccga
     9181 atgaccaaac taaatgtggt caatgaagcc atagaatata ttcccaagaa aaccaaagag
     9241 gggaatccgg ttacggtacg tgttccactt aacgacaaag cgaaagaaat ccttgaacgc
     9301 tacaaagaat atgagggaaa actgttgccg ttcatatccg agcaaaagta caatgatgcc
     9361 ataaaaaaga tattcaaatt agctggagtt gaccgcatcg taacaatctt agacccgttg
     9421 acgcacaacg aaatcaaacg acctatttat gaagtggcaa gcagccatct ggcaagacgt
     9481 acgtttatcg gcaatatcta taaaaaagtg aaagacccga accttgtttc cgcactgtcg
     9541 ggacacaagg agggaagcaa agctttcaga cgatacaggg atattgacga agaaatgaag
     9601 aaagaccttg taaaactact ggactgataa aagtatcaaa cagtttcata gatagtt
//
