LOCUS       sequence-313839-        7171 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bile Acid inducible sgRNA-2 with PM2 driving Nanoluc expression (NOT
            Gate 2), pNBU1 backbone, AmpR, ErmR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7171)
  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 7171)
  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: pMT449
FEATURES             Location/Qualifiers
     source          1..7171
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1410..1478,1479..2270)
                     /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(1628..1647)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            2401..2510
                     /note="incP origin of transfer"
     rep_origin      2681..3069
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     misc_RNA        3302..3377
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        3599..3648
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
ORIGIN
        1 tatagcagta agtcattatt tcttttggtt gaacgtagag agtagcgata ttaaaagaat
       61 ccgatgagaa aagactaata tttatctatc cattcagttt gatttctcag gactttacat
      121 cgtcctgaaa gtatttgttg gtaccgagga cgcgtaaaca tttacagttg catgtggcct
      181 attgttttta gccgttaaat attttataac tattaaatag cgatacaaat tgttcgaaac
      241 taatattgtt tatatcatat attctcgcat gttttaaagc tttattaaat tgattttttg
      301 taaacagttt ttcgtactct ttgttaaccc atttcattac aaaagtttca tatttttttc
      361 tctctttaaa tgccattttt gctggctttc tttttaatac aattaatgtg ctatccactt
      421 taggttttgg atggaaataa tacctaggaa tttttgctaa tatagaaata tctacctctg
      481 ccattaacag caatgctagt gatctgtttg tatctaataa cattttagca aaaccatatt
      541 ccactattaa ataacttatt gtggctgaac tttcaaaaac aatttttcga attatatttg
      601 tgcttatgtt gtaaggtatg ctgccaaata ttttatatgg attgtggcta ggaaatgtaa
      661 atttcagtat atcatcattt actatttgat agttaggata atttaagagc ttattacgag
      721 ttacctcaca taatttagaa tcaatttcta tcgccgttac aaaattacat ctctttacca
      781 atccagcagt aaaatgacct ttccctgcac ctatttcaaa gatgttatct ttttcatcta
      841 aacttatgca attcattatt ttttctatgt gatattttga agtaataaaa ttttgactat
      901 cttttatatt tactttgttc attataacct ctccttaatt tattgcatct cttttcgaat
      961 atttatgttt tttgagaaaa gaacgtactc atggttcatc ccgatatgcg tatcggtctg
     1021 tatatcagca actttctatg tgtttcaact acaatagtca tctattctca tctttctgag
     1081 tccaccccct gcaaagcccc tctttacgac ataaaaattc ggtcggaaaa ggtatgcaaa
     1141 agatgtttct ctctttaaga gaaactcttc gggatgcaaa aatatgaaaa taactccaat
     1201 tcaccaaatt atatagcgac ttttttacaa aatgctaaaa tttgttgatt tccgtcaagc
     1261 aattgttgag caaaaatgtc ttttacgata aaatgatacc tcaatatcaa ctgtttagca
     1321 aaacgatatt tctcttaaag agagaaacac ctttttgttc accaatcccc gacttttaat
     1381 cccgcggcca tgattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     1441 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     1501 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     1561 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     1621 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     1681 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     1741 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     1801 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     1861 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     1921 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     1981 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     2041 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     2101 ataaatctgg agccggtgag cgtgggtcac gcggtatcat tgcagcactg gggccagatg
     2161 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     2221 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     2281 aagtttactc ataacgcgtc aattcgaggg ggatcaattc cgtgataggt gggctgccct
     2341 tcctggttgg cttggtttca tcagccatcc gcttgccctc atctgttacg ccggcggtag
     2401 ccggccagcc tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa taagggacag
     2461 tgaagaagga acacccgctc gcgggtgggc ctacttcacc tatcctgccc ggctgacgcc
     2521 gttggataca ccaaggaaag tctacacgaa ccctttggca aaatcctgta tatcgtgcga
     2581 aaaaggatgg atataccgaa aaaatcgcta taatgacccc gaagcagggt tatgcagcgg
     2641 aaaacggaat tgatccggcc acgatgcgtc cggcgtagag gatctgaaga tcagcagttc
     2701 aacctgttga tagtacgtac taagctctca tgtttcacgt actaagctct catgtttaac
     2761 gtactaagct ctcatgttta acgaactaaa ccctcatggc taacgtacta agctctcatg
     2821 gctaacgtac taagctctca tgtttcacgt actaagctct catgtttgaa caataaaatt
     2881 aatataaatc agcaacttaa atagcctcta aggttttaag ttttataaga aaaaaaagaa
     2941 tatataaggc ttttaaagct tttaaggttt aacggttgtg gacaacaagc cagggatgta
     3001 acgcactgag aagcccttag agcctctcaa agcaattttg agtgacacag gaacacttaa
     3061 cggctgacat gggaattccc ctccaccgcg gtgtattaat gcggctgcaa tttttttggg
     3121 cggggccgcc caaaaaaatc ctagcaccct gcagggtctc ccgctttaca aagaaaattc
     3181 gacaaactgt tatttttcta tctatttaaa tgtacacccg tttgtttact ttactttagt
     3241 tatgtacctt tgtcggcaat gtacacccga aagtttactt tcaaagataa atcagagaaa
     3301 agttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag
     3361 tggcaccgag tcggtgcttt ttttggatcc ttcagtcaaa aaacttaaga ccgccggtct
     3421 tgtccactac cttgcagtaa tgcggtggac aggatcggcg gttttctttt ctcttctcaa
     3481 gcggccgctg ctgtcatagg ccggttgtcc ggtagtccgg atgatttaag gaagatttct
     3541 cttctttttt cactttctcc gaacaacctc cattttctct gatttatctt tgcagtagct
     3601 gtcaccggat gtgctttccg gtctgatgag tccgtgagga cgaaacagcc tctacaaata
     3661 attttgttta acgcatttta aaataaaata aattatttat ttaattaaac gaatatggtt
     3721 tttactctgg aagattttgt tggcgattgg cgtcagaccg cgggttataa tttggatcaa
     3781 gtcctggaac agggtggcgt aagctctctg ttccagaacc tgggtgtgag cgtgacgccg
     3841 attcagcgca tcgttctgtc cggcgagaac ggtctgaaaa ttgatattca tgtgatcatc
     3901 ccgtacgaag gcctgagcgg tgaccaaatg ggtcaaatcg agaaaatctt taaagtcgtc
     3961 tacccagttg acgatcacca cttcaaggtt atcttgcatt acggtacgct ggtgattgat
     4021 ggtgtgaccc cgaatatgat tgactatttc ggccgtccgt atgaaggcat tgccgttttt
     4081 gacggtaaaa agatcaccgt caccggtacc ctgtggaatg gcaataagat tattgacgag
     4141 cgtctgatta acccggacgg cagcctgctg ttccgcgtga ccatcaacgg tgtcacgggt
     4201 tggcgtctgt gcgagcgcat cctggcataa tgaactggga tccctcggta ccaaattttc
     4261 gaaaaaagac gctgaaaagc gtcttttttc gttttggtcc gcggccgccg gccgctgatc
     4321 tggaagaagc aatgaaagct gctgttaagt ctccgaatca ggtattgttc ctgacaggtg
     4381 tattcccatc cggtaaacgc ggatactttg cagttgatct gactcaggaa taaattataa
     4441 attaaggtaa gaagattgta ggataagcta atgaaataga aaaaggatgc cgtcacacaa
     4501 cttgtcggca ttcttttttg ttttattagt tgaaaatata gtgaaaaagt tgcctaaata
     4561 tgtatgttaa caaattattt gtcgtaactt tgcactccaa atctgttttt aacatatggc
     4621 actaatgcag aaaaaattga ctcgttctca gcagaaacac ttggacatca tcaacgctgc
     4681 taaagaagaa ttcatcgaat tcggtttctt ggctgctaac atggaccgta tcacttcttc
     4741 tgctgaagta tctaaacgta ctttgtaccg tcacttcgaa tctaaagaag tattgttcga
     4801 atctgtattg actatcatca acgactctgt aaacgaatct atctcttacc acttcgaccc
     4861 gaacaaatct actgaagaac agttgactga aatcgcttac aaagaaatcg acgtattgta
     4921 caaaacttac ggtatcgctt tggctcgtac tatcgtaatg gaattcttgc gtcagccgga
     4981 aatggctaaa actttgatcc agaacatcta ctctatccgt gctatcactc agtggttccg
     5041 ttctgctatc gaagctaaac gtttgaaaga cgctgacccg aaattgatga ctgacgtata
     5101 cgtatctttg ttccagggtt tgttcttctg gccgcaggta atgcacttgg acttggaacc
     5161 gcacggtgaa gaattgtctc agaaaatcga aactttgact actatcttct tgcagtctta
     5221 cggtgtagct gaataacaat gaatgggctt tcctcggtac caaattccag aaaagaggcc
     5281 tcccgaaagg ggggcctttt ttcgttttgg tccctaattg cctatcttcc agtgatggaa
     5341 cagcatttgt gcattggctg caacaatcag ccttacttgt gcctgttcta tttccgaacc
     5401 gaccgcttgt atgaatgcat caaaattcgt tttctctacg ttggattcct tgttgctcat
     5461 attgtgatga taatttctac aaatatagtc attggtaact atctatgaaa ctgtttgata
     5521 cttttatcag tccagtagtt ttacaaggtc tttcttcatt tcttcgtcaa tatccctgta
     5581 tcgtctgaaa gctttgcttc cctccttgtg tcccgacagt gcggaaacaa ggttcgggtc
     5641 tttcactttt ttatagatat tgccgataaa cgtacgtctt gccagatggc tgcttgccac
     5701 ttcataaata ggtcgtttga tttcgttgtg cgtcaacggg tctaagattg ttacgatgcg
     5761 gtcaactcca gctaatttga atatcttttt tatggcatca ttgtactttt gctcggatat
     5821 gaacggcaac agttttccct catattcttt gtagcgttca aggatttctt tcgctttgtc
     5881 gttaagtgga acacgtaccg taaccggatt cccctctttg gttttcttgg gaatatattc
     5941 tatggcttca ttgaccacat ttagtttggt cattcggtac aggtcgctca ccctgcatcc
     6001 tatcagtgtc tgaaatatga atatatccct ctgtattgcc agttgtgggg tggcagaaag
     6061 gtctgcatta aaaatcctgt ccctttcttc gagtgttata taataaggtg taccatatgt
     6121 acactcctct atcggaaact tgtcgaaagg tctgtttgtg gtgcgtttgt tatcgaagca
     6181 ccacaggaag aatgtgcgta ttcttgaaaa acagtctatc agcgtgtttt tgcttctggg
     6241 ctgtggtgtc ctcttttcgg gaatggcttc ataaatgctc gggtaaagtt cataatactg
     6301 gtattcgttc tgaaagaaat cccacatatc ccgaagcgtg tcaggtgtta ccaaatccac
     6361 atcaaggata aagccctttt gtcctctctt tgtagccctt acatatagtt cataacgcag
     6421 taaggctctt ttgacaaccc ggaaattttt ctttcgtact tccgacaaag ggtgcttgtt
     6481 tagaaattca tcgaatagtt ctccaatggt aggcttgata accacttcct ccggcaaaaa
     6541 atatttttca ggatggtaaa atttatcaag tgttgttttt agccattctt tgtctattgc
     6601 ttccttttct tgttgatata ccttctcgat ataggttttc aactggcgta tctcttcatt
     6661 tatatgggta cgcatttctt cattgcagac agctttcgtt tttacacatt cgtctttatc
     6721 atcccataga ttgggattga tggctaactg agtaggagca acggagtcta actgccttcc
     6781 gtttctaaaa cggacataga ttgtagccat ggattctgta tcatatcgtt tggctgcttt
     6841 tttaatgata aaggttactt tcatagactt tcaggttgaa ttttactctg ctgcaaatat
     6901 aaatattttc cccagcattt tccccacatc tgctaaatat tttgcaattc gattaaactt
     6961 ggattaaaat ttaataggat tataacatat tgaaatacag tgtaattgtg gcgtttttcc
     7021 gcatttttct ttttacccac attttcccca ctttaattat ttcaaatgcg gggtctgggt
     7081 acaagaaaga aagctaagta tttgatagtt caatacttag ctttttcttt tgcttgaatt
     7141 ttccccacat tttccccaca cgtgcaaaaa a
//
