LOCUS       sequence-313937-        9337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nanoluc placed on Output 1 of the 2in3out circuit (A001 and P002 
            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 9337)
  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 9337)
  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: pMT469
FEATURES             Location/Qualifiers
     source          1..9337
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    116..134
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    178..196
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        221..296
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    457..475
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    519..537
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        562..637
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        848..923
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        1130..1205
                     /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"
     misc_RNA        1714..1789
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        1977..2026
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     misc_RNA        2298..2347
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     misc_RNA        3129..3178
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             join(7605..7673,7674..8465)
                     /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(7823..7842)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            8596..8705
                     /note="incP origin of transfer"
     rep_origin      8876..9264
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 agcaccctgc agggtctccc gctggagttt tgcacccgct ttccaagaga agaaagcctt
       61 gttaaattga cttagtgtaa aagcgcagta ctgcttgacc ataagaacaa aaaaatctct
      121 atcactgata gggataaagt ttggaagata aagctaaaag ttcttatctt tgcagtctcc
      181 ctatcagtga tagagacgca caaagataat agaacttaag gttttagagc tagaaatagc
      241 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
      301 tttggatcca aaaaaaaaaa acaccctaac gggtgttttt ttttttttgg tctgccgcgg
      361 ccgctacgtt ttgcacccgc tttccaagag aagaaagcct tgttaaattg acttagtgta
      421 aaagcgcagt actgcttgac cataagaaca aaaaaatctc tatcactgat agggataaag
      481 tttggaagat aaagctaaaa gttcttatct ttgcagtctc cctatcagtg atagagacgc
      541 acaaagataa ttaagttaag ggttttagag ctagaaatag caagttaaaa taaggctagt
      601 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttggatcc ctcggtacca
      661 aattccagaa aagagacgct gaaaagcgtc ttttttcgtt ttggtccgcg gccgcaggtt
      721 gctgtcatag gccggttgtc cggtagtccg gatgatttaa ggaagatttc tcttcttttt
      781 tcactttctc cgaacaacct ccacttaagt tctattatct ttgcagtcaa agatacttat
      841 gttagaagtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg
      901 aaaaagtggc accgagtcgg tgcttttttt ggatccaacg catgagaaag cccccggaag
      961 atcaccttcc gggggctttt ttattgcgcg cggccgcgct ttgctgtcat aggccggttg
     1021 tccggtagtc cggatgattt aaggaagatt tctcttcttt tttcactttc tccgaacaac
     1081 ctccacttaa gttctattat ctttgcagtc aaagataatt aagttaaggg ttttagagct
     1141 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
     1201 ggtgcttttt ttggatccct cggtaccaaa ttccagaaaa gagacgctga aaagcgtctt
     1261 ttttcgtttt ggtccgcggc cgccgcttta caaagaaaat tcgacaaact gttatttttc
     1321 tatctattta aatgtacacc cgtttgttta ctttacttta gttatgtacc tttgtcggca
     1381 atgtacaccc gaaagtttac tttcaaagat acttatgtta gaagttttag agctagaaat
     1441 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
     1501 ttttttggat ccaacgcatg agaaagcccc cggaagatca ccttccgggg gcttttttat
     1561 tgcgcgcggc cgctgtctta caaagaaaat tcgacaaact gttatttttc tatctattta
     1621 aatgtacacc cgtttgttta ctttacttta gttatgtacc tttgtcggca atgtacaccc
     1681 gaaagtttac tttcaaagat aatagaactt aaggttttag agctagaaat agcaagttaa
     1741 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct ttttttggat
     1801 ccaaaaaaaa aaaacaccct aacgggtgtt tttttttttt ttggtctgcc gcggccgcaa
     1861 tgggagtgct gtcataggcc ggttgtccgg tagtccggat gatttaagga agatttctct
     1921 tcttttttca ctttctccga acaacctcca ttctaacata agtatctttg cagtagctgt
     1981 caccggatgt gctttccggt ctgatgagtc cgtgaggacg aaacagcctc tacaaataat
     2041 tttgtttaac gcattttaaa ataaaataaa ttatttattt aattaaacga atatgtaatg
     2101 aactgggatc cctcggtacc aaattttcga aaaaagacgc tgaaaagcgt cttttttcgt
     2161 tttggtccgc ggccgccggc cgctacgtgc tgtcataggc cggttgtccg gtagtccgga
     2221 tgatttaagg aagatttctc ttcttttttc actttctccg aacaacctcc acttaagttc
     2281 tattatcttt gcagtagctg tcaccggatg tgctttccgg tctgatgagt ccgtgaggac
     2341 gaaacagcct ctacaaataa ttttgtttaa cgcattttaa aataaaataa attatttatt
     2401 taattaaacg aatatggttt ttactctgga agattttgtt ggcgattggc gtcagaccgc
     2461 gggttataat ttggatcaag tcctggaaca gggtggcgta agctctctgt tccagaacct
     2521 gggtgtgagc gtgacgccga ttcagcgcat cgttctgtcc ggcgagaacg gtctgaaaat
     2581 tgatattcat gtgatcatcc cgtacgaagg cctgagcggt gaccaaatgg gtcaaatcga
     2641 gaaaatcttt aaagtcgtct acccagttga cgatcaccac ttcaaggtta tcttgcatta
     2701 cggtacgctg gtgattgatg gtgtgacccc gaatatgatt gactatttcg gccgtccgta
     2761 tgaaggcatt gccgtttttg acggtaaaaa gatcaccgtc accggtaccc tgtggaatgg
     2821 caataagatt attgacgagc gtctgattaa cccggacggc agcctgctgt tccgcgtgac
     2881 catcaacggt gtcacgggtt ggcgtctgtg cgagcgcatc ctggcataat gaactgggat
     2941 ccctcggtac caaattttcg aaaaaagacg ctgaaaagcg tcttttttcg ttttggtccg
     3001 cggccgccgg ccgcaggttg ctgtcatagg ccggttgtcc ggtagtccgg atgatttaag
     3061 gaagatttct cttctttttt cactttctcc gaacaacctc caccttaact taattatctt
     3121 tgcagtagct gtcaccggat gtgctttccg gtctgatgag tccgtgagga cgaaacagcc
     3181 tctacaaata attttgttta acgcatttta aaataaaata aattatttat ttaattaaac
     3241 gaatatgtaa tgaactggga tccctcggta ccaaattttc gaaaaaagac gctgaaaagc
     3301 gtcttttttc gttttggtcc gcggccgccg gccgcaatgt gatctggaag aagcaatgaa
     3361 agctgctgtt aagtctccga atcaggtatt gttcctgaca ggtgtattcc catccggtaa
     3421 acgcggatac tttgcagttg atctgactca ggaataaatt ataaattaag gtaagaagat
     3481 tgtaggataa gctaatgaaa tagaaaaagg atgccgtcac acaacttgtc ggcattcttt
     3541 tttgttttat tagttgaaaa tatagtgaaa aagttgccta aatatgtatg ttaacaaatt
     3601 atttgtcgta actttgcact ccaaatctgt ttttaacata tggcactaat gcagaaaaaa
     3661 ttgactcgtt ctcagcagaa acacttggac atcatcaacg ctgctaaaga agaattcatc
     3721 gaattcggtt tcttggctgc taacatggac cgtatcactt cttctgctga agtatctaaa
     3781 cgtactttgt accgtcactt cgaatctaaa gaagtattgt tcgaatctgt attgactatc
     3841 atcaacgact ctgtaaacga atctatctct taccacttcg acccgaacaa atctactgaa
     3901 gaacagttga ctgaaatcgc ttacaaagaa atcgacgtat tgtacaaaac ttacggtatc
     3961 gctttggctc gtactatcgt aatggaattc ttgcgtcagc cggaaatggc taaaactttg
     4021 atccagaaca tctactctat ccgtgctatc actcagtggt tccgttctgc tatcgaagct
     4081 aaacgtttga aagacgctga cccgaaattg atgactgacg tatacgtatc tttgttccag
     4141 ggtttgttct tctggccgca ggtaatgcac ttggacttgg aaccgcacgg tgaagaattg
     4201 tctcagaaaa tcgaaacttt gactactatc ttcttgcagt cttacggtgt agctgaataa
     4261 caatgaatgg gctttcctcg gtaccaaatt ccagaaaaga ggcctcccga aaggggggcc
     4321 ttttttcgtt ttggtcccta attgcctatc ttccagtgat ggaacagcat ttgtgcattg
     4381 gctgcaacaa tcagccttac ttgtgcctgt tctatttccg aaccgaccgc ttgtatgaat
     4441 gcatcaaaat tcgttttctc tacgttggat tccttgttgc tcatattgtg atgataattt
     4501 ctacaaatat agtcattggt aactatctat gaaactgttt gatactttta tcagtccagt
     4561 agttttacaa ggtctttctt catttcttcg tcaatatccc tgtatcgtct gaaagctttg
     4621 cttccctcct tgtgtcccga cagtgcggaa acaaggttcg ggtctttcac ttttttatag
     4681 atattgccga taaacgtacg tcttgccaga tggctgcttg ccacttcata aataggtcgt
     4741 ttgatttcgt tgtgcgtcaa cgggtctaag attgttacga tgcggtcaac tccagctaat
     4801 ttgaatatct tttttatggc atcattgtac ttttgctcgg atatgaacgg caacagtttt
     4861 ccctcatatt ctttgtagcg ttcaaggatt tctttcgctt tgtcgttaag tggaacacgt
     4921 accgtaaccg gattcccctc tttggttttc ttgggaatat attctatggc ttcattgacc
     4981 acatttagtt tggtcattcg gtacaggtcg ctcaccctgc atcctatcag tgtctgaaat
     5041 atgaatatat ccctctgtat tgccagttgt ggggtggcag aaaggtctgc attaaaaatc
     5101 ctgtcccttt cttcgagtgt tatataataa ggtgtaccat atgtacactc ctctatcgga
     5161 aacttgtcga aaggtctgtt tgtggtgcgt ttgttatcga agcaccacag gaagaatgtg
     5221 cgtattcttg aaaaacagtc tatcagcgtg tttttgcttc tgggctgtgg tgtcctcttt
     5281 tcgggaatgg cttcataaat gctcgggtaa agttcataat actggtattc gttctgaaag
     5341 aaatcccaca tatcccgaag cgtgtcaggt gttaccaaat ccacatcaag gataaagccc
     5401 ttttgtcctc tctttgtagc ccttacatat agttcataac gcagtaaggc tcttttgaca
     5461 acccggaaat ttttctttcg tacttccgac aaagggtgct tgtttagaaa ttcatcgaat
     5521 agttctccaa tggtaggctt gataaccact tcctccggca aaaaatattt ttcaggatgg
     5581 taaaatttat caagtgttgt ttttagccat tctttgtcta ttgcttcctt ttcttgttga
     5641 tataccttct cgatataggt tttcaactgg cgtatctctt catttatatg ggtacgcatt
     5701 tcttcattgc agacagcttt cgtttttaca cattcgtctt tatcatccca tagattggga
     5761 ttgatggcta actgagtagg agcaacggag tctaactgcc ttccgtttct aaaacggaca
     5821 tagattgtag ccatggattc tgtatcatat cgtttggctg cttttttaat gataaaggtt
     5881 actttcatag actttcaggt tgaattttac tctgctgcaa atataaatat tttccccagc
     5941 attttcccca catctgctaa atattttgca attcgattaa acttggatta aaatttaata
     6001 ggattataac atattgaaat acagtgtaat tgtggcgttt ttccgcattt ttctttttac
     6061 ccacattttc cccactttaa ttatttcaaa tgcggggtct gggtacaaga aagaaagcta
     6121 agtatttgat agttcaatac ttagcttttt cttttgcttg aattttcccc acattttccc
     6181 cacacgtgca aaaaatatag cagtaagtca ttatttcttt tggttgaacg tagagagtag
     6241 cgatattaaa agaatccgat gagaaaagac taatatttat ctatccattc agtttgattt
     6301 ctcaggactt tacatcgtcc tgaaagtatt tgttggtacc gaggacgcgt aaacatttac
     6361 agttgcatgt ggcctattgt ttttagccgt taaatatttt ataactatta aatagcgata
     6421 caaattgttc gaaactaata ttgtttatat catatattct cgcatgtttt aaagctttat
     6481 taaattgatt ttttgtaaac agtttttcgt actctttgtt aacccatttc attacaaaag
     6541 tttcatattt ttttctctct ttaaatgcca tttttgctgg ctttcttttt aatacaatta
     6601 atgtgctatc cactttaggt tttggatgga aataatacct aggaattttt gctaatatag
     6661 aaatatctac ctctgccatt aacagcaatg ctagtgatct gtttgtatct aataacattt
     6721 tagcaaaacc atattccact attaaataac ttattgtggc tgaactttca aaaacaattt
     6781 ttcgaattat atttgtgctt atgttgtaag gtatgctgcc aaatatttta tatggattgt
     6841 ggctaggaaa tgtaaatttc agtatatcat catttactat ttgatagtta ggataattta
     6901 agagcttatt acgagttacc tcacataatt tagaatcaat ttctatcgcc gttacaaaat
     6961 tacatctctt taccaatcca gcagtaaaat gacctttccc tgcacctatt tcaaagatgt
     7021 tatctttttc atctaaactt atgcaattca ttattttttc tatgtgatat tttgaagtaa
     7081 taaaattttg actatctttt atatttactt tgttcattat aacctctcct taatttattg
     7141 catctctttt cgaatattta tgttttttga gaaaagaacg tactcatggt tcatcccgat
     7201 atgcgtatcg gtctgtatat cagcaacttt ctatgtgttt caactacaat agtcatctat
     7261 tctcatcttt ctgagtccac cccctgcaaa gcccctcttt acgacataaa aattcggtcg
     7321 gaaaaggtat gcaaaagatg tttctctctt taagagaaac tcttcgggat gcaaaaatat
     7381 gaaaataact ccaattcacc aaattatata gcgacttttt tacaaaatgc taaaatttgt
     7441 tgatttccgt caagcaattg ttgagcaaaa atgtctttta cgataaaatg atacctcaat
     7501 atcaactgtt tagcaaaacg atatttctct taaagagaga aacacctttt tgttcaccaa
     7561 tccccgactt ttaatcccgc ggccatgatt gaaaaaggaa gagtatgagt attcaacatt
     7621 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     7681 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     7741 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     7801 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     7861 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     7921 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     7981 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     8041 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     8101 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     8161 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     8221 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     8281 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtcacgcggt atcattgcag
     8341 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     8401 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     8461 ggtaactgtc agaccaagtt tactcataac gcgtcaattc gagggggatc aattccgtga
     8521 taggtgggct gcccttcctg gttggcttgg tttcatcagc catccgcttg ccctcatctg
     8581 ttacgccggc ggtagccggc cagcctcgca gagcaggatt cccgttgagc accgccaggt
     8641 gcgaataagg gacagtgaag aaggaacacc cgctcgcggg tgggcctact tcacctatcc
     8701 tgcccggctg acgccgttgg atacaccaag gaaagtctac acgaaccctt tggcaaaatc
     8761 ctgtatatcg tgcgaaaaag gatggatata ccgaaaaaat cgctataatg accccgaagc
     8821 agggttatgc agcggaaaac ggaattgatc cggccacgat gcgtccggcg tagaggatct
     8881 gaagatcagc agttcaacct gttgatagta cgtactaagc tctcatgttt cacgtactaa
     8941 gctctcatgt ttaacgtact aagctctcat gtttaacgaa ctaaaccctc atggctaacg
     9001 tactaagctc tcatggctaa cgtactaagc tctcatgttt cacgtactaa gctctcatgt
     9061 ttgaacaata aaattaatat aaatcagcaa cttaaatagc ctctaaggtt ttaagtttta
     9121 taagaaaaaa aagaatatat aaggctttta aagcttttaa ggtttaacgg ttgtggacaa
     9181 caagccaggg atgtaacgca ctgagaagcc cttagagcct ctcaaagcaa ttttgagtga
     9241 cacaggaaca cttaacggct gacatgggaa ttcccctcca ccgcggtgta ttaatgcggc
     9301 tgcaattttt ttgggcgggg ccgcccaaaa aaatcct
//
