LOCUS       sequence-226365-        5551 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration vector at an attB site in Bacteroides; contains the 
            P1T_DP aTC-inducible promoter with the RBS GH023 upstream of the 
            NanoLuc reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5551)
  AUTHORS   Lim B, Zimmermann M, Barry NA, Goodman AL
  TITLE     Engineered Regulatory Systems Modulate Gene Expression of Human 
            Commensals in the Gut.
  JOURNAL   Cell. 2017 Apr 20;169(3):547-558.e15. doi: 
            10.1016/j.cell.2017.03.045.
  PUBMED    28431252
REFERENCE   2  (bases 1 to 5551)
  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: 
            "10.1016/j.cell.2017.03"; journalName: "Cell"; date: "2017-04-20- 
            20"; volume: "169"; issue: "3"; pages: "547-558.e15"
COMMENT     Sequence Label: pNBU2_erm_P1T_DP-GH023 - NanoLuc
FEATURES             Location/Qualifiers
     source          1..5551
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            102..211
                     /note="incP origin of transfer"
     rep_origin      382..770
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    885..903
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    947..965
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             997..1512
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             join(4662..4730,4731..5522)
                     /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(4880..4899)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 caattcgagg gggatcaatt ccgtgatagg tgggctgccc ttcctggttg gcttggtttc
       61 atcagccatc cgcttgccct catctgttac gccggcggta gccggccagc ctcgcagagc
      121 aggattcccg ttgagcaccg ccaggtgcga ataagggaca gtgaagaagg aacacccgct
      181 cgcgggtggg cctacttcac ctatcctgcc cggctgacgc cgttggatac accaaggaaa
      241 gtctacacga accctttggc aaaatcctgt atatcgtgcg aaaaaggatg gatataccga
      301 aaaaatcgct ataatgaccc cgaagcaggg ttatgcagcg gaaaacggaa ttgatccggc
      361 cacgatgcgt ccggcgtaga ggatctgaag atcagcagtt caacctgttg atagtacgta
      421 ctaagctctc atgtttcacg tactaagctc tcatgtttaa cgtactaagc tctcatgttt
      481 aacgaactaa accctcatgg ctaacgtact aagctctcat ggctaacgta ctaagctctc
      541 atgtttcacg tactaagctc tcatgtttga acaataaaat taatataaat cagcaactta
      601 aatagcctct aaggttttaa gttttataag aaaaaaaaga atatataagg cttttaaagc
      661 ttttaaggtt taacggttgt ggacaacaag ccagggatgt aacgcactga gaagccctta
      721 gagcctctca aagcaatttt gagtgacaca ggaacactta acggctgaca tgggaattcc
      781 cctccaccgc ggtggctttt gcacccgctt tccaagagaa gaaagccttg ttaaattgac
      841 ttagtgtaaa agcgcagtac tgcttgacca taagaacaaa aaaatctcta tcactgatag
      901 ggataaagtt tggaagataa agctaaaagt tcttatcttt gcagtctccc tatcagtgat
      961 agagacgaaa taaagacata taaaagaaaa gacaccatgg tcttcacact cgaagatttc
     1021 gttggggact ggcgacagac agccggctac aacctggacc aagtccttga acagggaggt
     1081 gtgtccagtt tgtttcagaa tctcggggtg tccgtaactc cgatccaaag gattgtcctg
     1141 agcggtgaaa atgggctgaa gatcgacatc catgtcatca tcccgtatga aggtctgagc
     1201 ggcgaccaaa tgggccagat cgaaaaaatt tttaaggtgg tgtaccctgt ggatgatcat
     1261 cactttaagg tgatcctgca ctatggcaca ctggtaatcg acggggttac gccgaacatg
     1321 atcgactatt tcggacggcc gtatgaaggc atcgccgtgt tcgacggcaa aaagatcact
     1381 gtaacaggga ccctgtggaa cggcaacaaa attatcgacg agcgcctgat caaccccgac
     1441 ggctccctgc tgttccgagt aaccatcaac ggagtgaccg gctggcggct gtgcgaacgc
     1501 attctggcgt aactaattgc ctatcttcca gtgatggaac agcatttgtg cattggctgc
     1561 aacaatcagc cttacttgtg cctgttctat ttccgaaccg accgcttgta tgaatgcatc
     1621 aaaattcgtt ttctctacgt tggattcctt gttgctcata ttgtgatgat aatttctaca
     1681 aatatagtca ttggtaacta tctatgaaac tgtttgatac ttttatagtt gattaaactt
     1741 gttcatggca tttgccttaa tatcatccgc tatgtcaatg tagggtttca tagctttgta
     1801 gtcgctgtgt cccgtccatt tcatgaccac ctgtgccggg attccgagag ccagcgcatt
     1861 gcagatgaat gtcctttttc ctgcatgggt actgagcaaa gcgtatttgg gtgtgacttc
     1921 atcaatacgt tcatttccct tgtagtaggt ttcccgtaca ggctcgttga tttctgccag
     1981 ttcgcccagc tctttcaggt aatcgttcat cttctggttg ctgatgacgg gcagagccat
     2041 gtaattctcg aaatggatgt ccttgtattt gtccagtatg gctttgctgt atttgttcag
     2101 ttcaatcgtc aggctgtcgg cagtcttgac tgtggttatt tcgatgtggt cggacttcac
     2161 atcgcttctt ttcagattgc gaacatccga ataccgcaaa ctcgtaaagc agcagaacag
     2221 gaaaacatca cgcacacgtt ccaggtattg cttatccttg ggtatctggt agtctttcag
     2281 cttgttcagt tcatcccaag tcaggaagat tacttttttc gaggtggttt tcagtttcgg
     2341 tttgaacgta tcgtatgcaa tgttctgatg atgtcctttc ttgaagctcc agcgcaggaa
     2401 ccatttgagg aatcccattt gcttgccgat ggtgctgttt ctcatatcct tggtgtcacg
     2461 caggaagttg acgtattcgt tcaatccaaa ctcgttgaaa tagttgaacg ttgcatcctc
     2521 cttgaactct ttgaggtggt tcctcactgc tgcaaatttt tcataggtgg atgccgtcca
     2581 gttattctgg ttaccgcact cttttacaaa ctcatcgaac acctcccaaa agctgacagg
     2641 ggcttcttcc ggctgttctt cactggtgtc tttcattctc atgttgaaag cttccttcaa
     2701 ctgttgggtc gttggcatga cctcctgcac ctcaaattcc ttgaaaatat tctggatttc
     2761 ggcatagtat ttcagcaagt ccgtattgat ttcggctgca ctttgcttta gcttgttggt
     2821 acatccgttc tttacccgct gcttatctgc atcccatttg gctacgtcaa tccggtagcc
     2881 cgttgtaaac tcgatacgtt ggctggcaaa gatgacacgc atacggatgg gtacgttctc
     2941 tacgattggc acaccgttct ttttccggct ctccaatgca aaaatgatgt tgcgcttgat
     3001 attcataatt gggtgcgttt gaaattctac acccaaatat acacccaatt attgagatag
     3061 caaaagacat ttagaaacat ttacttttac tctatattgt aatttacact tgattatcag
     3121 tcgtttgcag ttttatgata ttctgtgaaa gtataagttc gagagcctgt ctctccgcaa
     3181 aaaacgctga aaatcagcag attgcaaaac aaacaccctg ttttacaccc aagaatgtaa
     3241 agtcgggtgt ttttgtttta tttaagataa tacaaccact acataataaa agagtagcga
     3301 tattaaaaga atccgatgag aaaagactaa tatttatcta tccattcagt ttgatttctc
     3361 aggactttac atcgtcctga aagtatttgt tggtaccgag gacgcgtaaa catttacagt
     3421 tgcatgtggc ctattgtttt tagccgttaa atattttata actattaaat agcgatacaa
     3481 attgttcgaa actaatattg tttatatcat atattctcgc atgttttaaa gctttattaa
     3541 attgattttt tgtaaacagt ttttcgtact ctttgttaac ccatttcatt acaaaagttt
     3601 catatttttt tctctcttta aatgccattt ttgctggctt tctttttaat acaattaatg
     3661 tgctatccac tttaggtttt ggatggaaat aatacctagg aatttttgct aatatagaaa
     3721 tatctacctc tgccattaac agcaatgcta gtgatctgtt tgtatctaat aacattttag
     3781 caaaaccata ttccactatt aaataactta ttgtggctga actttcaaaa acaatttttc
     3841 gaattatatt tgtgcttatg ttgtaaggta tgctgccaaa tattttatat ggattgtggc
     3901 taggaaatgt aaatttcagt atatcatcat ttactatttg atagttagga taatttaaga
     3961 gcttattacg agttacctca cataatttag aatcaatttc tatcgccgtt acaaaattac
     4021 atctctttac caatccagca gtaaaatgac ctttccctgc acctatttca aagatgttat
     4081 ctttttcatc taaacttatg caattcatta ttttttctat gtgatatttt gaagtaataa
     4141 aattttgact atcttttata tttactttgt tcattataac ctctccttaa tttattgcat
     4201 ctcttttcga atatttatgt tttttgagaa aagaacgtac tcatggttca tcccgatatg
     4261 cgtatcggtc tgtatatcag caactttcta tgtgtttcaa ctacaatagt catctattct
     4321 catctttctg agtccacccc ctgcaaagcc cctctttacg acataaaaat tcggtcggaa
     4381 aaggtatgca aaagatgttt ctctctttaa gagaaactct tcgggatgca aaaatatgaa
     4441 aataactcca attcaccaaa ttatatagcg acttttttac aaaatgctaa aatttgttga
     4501 tttccgtcaa gcaattgttg agcaaaaatg tcttttacga taaaatgata cctcaatatc
     4561 aactgtttag caaaacgata tttctcttaa agagagaaac acctttttgt tcaccaatcc
     4621 ccgactttta atcccgcggc catgattgaa aaaggaagag tatgagtatt caacatttcc
     4681 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4741 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4801 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4861 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4921 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4981 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5041 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5101 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5161 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5221 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5281 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5341 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5401 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5461 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5521 aactgtcaga ccaagtttac tcataacgcg t
//
