LOCUS       sequence-313188-        5606 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LacZ, pNBU1 backbone with BsaI sites, AmpR, ErmR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5606)
  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 5606)
  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: pMT405
FEATURES             Location/Qualifiers
     source          1..5606
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     525..547
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     539..556
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     540..556
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    557..613
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(626..642)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(626..642)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(639..661)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    650..666
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(674..680,681..698,699..704))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    719..740
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             join(4180..4248,4249..5040)
                     /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(4398..4417)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            5171..5280
                     /note="incP origin of transfer"
     rep_origin      join(5451..5606,1..233)
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 ctcatgtttc acgtactaag ctctcatgtt tgaacaataa aattaatata aatcagcaac
       61 ttaaatagcc tctaaggttt taagttttat aagaaaaaaa agaatatata aggcttttaa
      121 agcttttaag gtttaacggt tgtggacaac aagccaggga tgtaacgcac tgagaagccc
      181 ttagagcctc tcaaagcaat tttgagtgac acaggaacac ttaacggctg acatgggaat
      241 tcccctccac cgcggtgtat taatgcggct gcaatttttt tgggcggggc cgcccaaaaa
      301 aatcctagca ccctgcaggg tctcccgctt ctgtcttctg caccatatgc ggtgtgaaat
      361 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattagccat tcaggctgcg
      421 caactgttgg gaagggcgat cggtgtgggc ctcttcgcta ttacgccagc tggcgaaagg
      481 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      541 taaaacgacg gccagtgaat tcgagctcgg tacccgggga tcctctagag tcgacctgca
      601 ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
      661 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
      721 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
      781 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gaagacaggg tctcccaatg
      841 aatgggcttt cctcggtacc aaattccaga aaagaggcct cccgaaaggg gggccttttt
      901 tcgttttggt ccctaattgc ctatcttcca gtgatggaac agcatttgtg cattggctgc
      961 aacaatcagc cttacttgtg cctgttctat ttccgaaccg accgcttgta tgaatgcatc
     1021 aaaattcgtt ttctctacgt tggattcctt gttgctcata ttgtgatgat aatttctaca
     1081 aatatagtca ttggtaacta tctatgaaac tgtttgatac ttttatcagt ccagtagttt
     1141 tacaaggtct ttcttcattt cttcgtcaat atccctgtat cgtctgaaag ctttgcttcc
     1201 ctccttgtgt cccgacagtg cggaaacaag gttcgggtct ttcacttttt tatagatatt
     1261 gccgataaac gtacgtcttg ccagatggct gcttgccact tcataaatag gtcgtttgat
     1321 ttcgttgtgc gtcaacgggt ctaagattgt tacgatgcgg tcaactccag ctaatttgaa
     1381 tatctttttt atggcatcat tgtacttttg ctcggatatg aacggcaaca gttttccctc
     1441 atattctttg tagcgttcaa ggatttcttt cgctttgtcg ttaagtggaa cacgtaccgt
     1501 aaccggattc ccctctttgg ttttcttggg aatatattct atggcttcat tgaccacatt
     1561 tagtttggtc attcggtaca ggtcgctcac cctgcatcct atcagtgtct gaaatatgaa
     1621 tatatccctc tgtattgcca gttgtggggt ggcagaaagg tctgcattaa aaatcctgtc
     1681 cctttcttcg agtgttatat aataaggtgt accatatgta cactcctcta tcggaaactt
     1741 gtcgaaaggt ctgtttgtgg tgcgtttgtt atcgaagcac cacaggaaga atgtgcgtat
     1801 tcttgaaaaa cagtctatca gcgtgttttt gcttctgggc tgtggtgtcc tcttttcggg
     1861 aatggcttca taaatgctcg ggtaaagttc ataatactgg tattcgttct gaaagaaatc
     1921 ccacatatcc cgaagcgtgt caggtgttac caaatccaca tcaaggataa agcccttttg
     1981 tcctctcttt gtagccctta catatagttc ataacgcagt aaggctcttt tgacaacccg
     2041 gaaatttttc tttcgtactt ccgacaaagg gtgcttgttt agaaattcat cgaatagttc
     2101 tccaatggta ggcttgataa ccacttcctc cggcaaaaaa tatttttcag gatggtaaaa
     2161 tttatcaagt gttgttttta gccattcttt gtctattgct tccttttctt gttgatatac
     2221 cttctcgata taggttttca actggcgtat ctcttcattt atatgggtac gcatttcttc
     2281 attgcagaca gctttcgttt ttacacattc gtctttatca tcccatagat tgggattgat
     2341 ggctaactga gtaggagcaa cggagtctaa ctgccttccg tttctaaaac ggacatagat
     2401 tgtagccatg gattctgtat catatcgttt ggctgctttt ttaatgataa aggttacttt
     2461 catagacttt caggttgaat tttactctgc tgcaaatata aatattttcc ccagcatttt
     2521 ccccacatct gctaaatatt ttgcaattcg attaaacttg gattaaaatt taataggatt
     2581 ataacatatt gaaatacagt gtaattgtgg cgtttttccg catttttctt tttacccaca
     2641 ttttccccac tttaattatt tcaaatgcgg ggtctgggta caagaaagaa agctaagtat
     2701 ttgatagttc aatacttagc tttttctttt gcttgaattt tccccacatt ttccccacac
     2761 gtgcaaaaaa tatagcagta agtcattatt tcttttggtt gaacgtagag agtagcgata
     2821 ttaaaagaat ccgatgagaa aagactaata tttatctatc cattcagttt gatttctcag
     2881 gactttacat cgtcctgaaa gtatttgttg gtaccgagga cgcgtaaaca tttacagttg
     2941 catgtggcct attgttttta gccgttaaat attttataac tattaaatag cgatacaaat
     3001 tgttcgaaac taatattgtt tatatcatat attctcgcat gttttaaagc tttattaaat
     3061 tgattttttg taaacagttt ttcgtactct ttgttaaccc atttcattac aaaagtttca
     3121 tatttttttc tctctttaaa tgccattttt gctggctttc tttttaatac aattaatgtg
     3181 ctatccactt taggttttgg atggaaataa tacctaggaa tttttgctaa tatagaaata
     3241 tctacctctg ccattaacag caatgctagt gatctgtttg tatctaataa cattttagca
     3301 aaaccatatt ccactattaa ataacttatt gtggctgaac tttcaaaaac aatttttcga
     3361 attatatttg tgcttatgtt gtaaggtatg ctgccaaata ttttatatgg attgtggcta
     3421 ggaaatgtaa atttcagtat atcatcattt actatttgat agttaggata atttaagagc
     3481 ttattacgag ttacctcaca taatttagaa tcaatttcta tcgccgttac aaaattacat
     3541 ctctttacca atccagcagt aaaatgacct ttccctgcac ctatttcaaa gatgttatct
     3601 ttttcatcta aacttatgca attcattatt ttttctatgt gatattttga agtaataaaa
     3661 ttttgactat cttttatatt tactttgttc attataacct ctccttaatt tattgcatct
     3721 cttttcgaat atttatgttt tttgagaaaa gaacgtactc atggttcatc ccgatatgcg
     3781 tatcggtctg tatatcagca actttctatg tgtttcaact acaatagtca tctattctca
     3841 tctttctgag tccaccccct gcaaagcccc tctttacgac ataaaaattc ggtcggaaaa
     3901 ggtatgcaaa agatgtttct ctctttaaga gaaactcttc gggatgcaaa aatatgaaaa
     3961 taactccaat tcaccaaatt atatagcgac ttttttacaa aatgctaaaa tttgttgatt
     4021 tccgtcaagc aattgttgag caaaaatgtc ttttacgata aaatgatacc tcaatatcaa
     4081 ctgtttagca aaacgatatt tctcttaaag agagaaacac ctttttgttc accaatcccc
     4141 gacttttaat cccgcggcca tgattgaaaa aggaagagta tgagtattca acatttccgt
     4201 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4261 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4321 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4381 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4441 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4501 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4561 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4621 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4681 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     4741 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     4801 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     4861 tttattgctg ataaatctgg agccggtgag cgtgggtcac gcggtatcat tgcagcactg
     4921 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4981 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5041 ctgtcagacc aagtttactc ataacgcgtc aattcgaggg ggatcaattc cgtgataggt
     5101 gggctgccct tcctggttgg cttggtttca tcagccatcc gcttgccctc atctgttacg
     5161 ccggcggtag ccggccagcc tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa
     5221 taagggacag tgaagaagga acacccgctc gcgggtgggc ctacttcacc tatcctgccc
     5281 ggctgacgcc gttggataca ccaaggaaag tctacacgaa ccctttggca aaatcctgta
     5341 tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta taatgacccc gaagcagggt
     5401 tatgcagcgg aaaacggaat tgatccggcc acgatgcgtc cggcgtagag gatctgaaga
     5461 tcagcagttc aacctgttga tagtacgtac taagctctca tgtttcacgt actaagctct
     5521 catgtttaac gtactaagct ctcatgttta acgaactaaa ccctcatggc taacgtacta
     5581 agctctcatg gctaacgtac taagct
//
