LOCUS       sequence-226485-        6357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration vector at an attB site in Bacteroides; contains 1kb 
            cassette of aTC-regulatable promoter (RBS GH023) upstream of the 
            NanoLuc gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6357)
  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 6357)
  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-TetR-P1T_DP-GH023 - NanoLuc
FEATURES             Location/Qualifiers
     source          1..6357
                     /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"
     CDS             complement(797..1420)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     1328..1347
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    1691..1709
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1753..1771
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1803..2318
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             join(5468..5536,5537..6328)
                     /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(5686..5705)
                     /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 ggtggcttaa gacccacttt cacatttaag ttgtttttct aatccgcata
      841 tgatcaattc aaggccgaat aagaaggctg gctctgcacc ttggtgatca aataattcga
      901 tagcttgtcg taataatggc ggcatactat cagtagtagg tgtttccctt tcttctttag
      961 cgacttgatg ctcttgatct tccaatacgc aacctaaagt aaaatgcccc acagcgctga
     1021 gtgcatataa tgcattctct agtgaaaaac cttgttggca taaaaaggct aattgatttt
     1081 cgagagtttc atactgtttt tctgtaggcc gtgtacctaa atgtactttt gctccatcgc
     1141 gatgacttag taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa tcttgccagc
     1201 tttccccttc taaagggcaa aagtgagtat ggtgcctatc taacatctca atggctaagg
     1261 cgtcgagcaa agcccgctta ttttttacat gccaatacaa tgtaggctgc tctacaccta
     1321 gcttctgggc gagtttacgg gttgttaaac cttcgattcc gacctcatta agcagctcta
     1381 atgcgctgtt aatcacttta cttttatcta atctagacat attcgtttaa tatcataaat
     1441 aatttatttt attttaaaat gcgcgggtgc aaaggtaaga ggttttattt taactaccaa
     1501 atgttttcgg aagttttttc gcttttcttt ttctatcgtt tctcagactc tcttagcgaa
     1561 agggaaagaa ggtaaagaag aaaaacaaaa cgccttttct ttttttgcac ccgctttcca
     1621 agagaagaaa gccttgttaa attgacttag tgtaaaagcg cagtactgct tgaccataag
     1681 aacaaaaaaa tctctatcac tgatagggat aaagtttgga agataaagct aaaagttctt
     1741 atctttgcag tctccctatc agtgatagag acgaaataaa gacatataaa agaaaagaca
     1801 ccatggtctt cacactcgaa gatttcgttg gggactggcg acagacagcc ggctacaacc
     1861 tggaccaagt ccttgaacag ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg
     1921 taactccgat ccaaaggatt gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg
     1981 tcatcatccc gtatgaaggt ctgagcggcg accaaatggg ccagatcgaa aaaattttta
     2041 aggtggtgta ccctgtggat gatcatcact ttaaggtgat cctgcactat ggcacactgg
     2101 taatcgacgg ggttacgccg aacatgatcg actatttcgg acggccgtat gaaggcatcg
     2161 ccgtgttcga cggcaaaaag atcactgtaa cagggaccct gtggaacggc aacaaaatta
     2221 tcgacgagcg cctgatcaac cccgacggct ccctgctgtt ccgagtaacc atcaacggag
     2281 tgaccggctg gcggctgtgc gaacgcattc tggcgtaact aattgcctat cttccagtga
     2341 tggaacagca tttgtgcatt ggctgcaaca atcagcctta cttgtgcctg ttctatttcc
     2401 gaaccgaccg cttgtatgaa tgcatcaaaa ttcgttttct ctacgttgga ttccttgttg
     2461 ctcatattgt gatgataatt tctacaaata tagtcattgg taactatcta tgaaactgtt
     2521 tgatactttt atagttgatt aaacttgttc atggcatttg ccttaatatc atccgctatg
     2581 tcaatgtagg gtttcatagc tttgtagtcg ctgtgtcccg tccatttcat gaccacctgt
     2641 gccgggattc cgagagccag cgcattgcag atgaatgtcc tttttcctgc atgggtactg
     2701 agcaaagcgt atttgggtgt gacttcatca atacgttcat ttcccttgta gtaggtttcc
     2761 cgtacaggct cgttgatttc tgccagttcg cccagctctt tcaggtaatc gttcatcttc
     2821 tggttgctga tgacgggcag agccatgtaa ttctcgaaat ggatgtcctt gtatttgtcc
     2881 agtatggctt tgctgtattt gttcagttca atcgtcaggc tgtcggcagt cttgactgtg
     2941 gttatttcga tgtggtcgga cttcacatcg cttcttttca gattgcgaac atccgaatac
     3001 cgcaaactcg taaagcagca gaacaggaaa acatcacgca cacgttccag gtattgctta
     3061 tccttgggta tctggtagtc tttcagcttg ttcagttcat cccaagtcag gaagattact
     3121 tttttcgagg tggttttcag tttcggtttg aacgtatcgt atgcaatgtt ctgatgatgt
     3181 cctttcttga agctccagcg caggaaccat ttgaggaatc ccatttgctt gccgatggtg
     3241 ctgtttctca tatccttggt gtcacgcagg aagttgacgt attcgttcaa tccaaactcg
     3301 ttgaaatagt tgaacgttgc atcctccttg aactctttga ggtggttcct cactgctgca
     3361 aatttttcat aggtggatgc cgtccagtta ttctggttac cgcactcttt tacaaactca
     3421 tcgaacacct cccaaaagct gacaggggct tcttccggct gttcttcact ggtgtctttc
     3481 attctcatgt tgaaagcttc cttcaactgt tgggtcgttg gcatgacctc ctgcacctca
     3541 aattccttga aaatattctg gatttcggca tagtatttca gcaagtccgt attgatttcg
     3601 gctgcacttt gctttagctt gttggtacat ccgttcttta cccgctgctt atctgcatcc
     3661 catttggcta cgtcaatccg gtagcccgtt gtaaactcga tacgttggct ggcaaagatg
     3721 acacgcatac ggatgggtac gttctctacg attggcacac cgttcttttt ccggctctcc
     3781 aatgcaaaaa tgatgttgcg cttgatattc ataattgggt gcgtttgaaa ttctacaccc
     3841 aaatatacac ccaattattg agatagcaaa agacatttag aaacatttac ttttactcta
     3901 tattgtaatt tacacttgat tatcagtcgt ttgcagtttt atgatattct gtgaaagtat
     3961 aagttcgaga gcctgtctct ccgcaaaaaa cgctgaaaat cagcagattg caaaacaaac
     4021 accctgtttt acacccaaga atgtaaagtc gggtgttttt gttttattta agataataca
     4081 accactacat aataaaagag tagcgatatt aaaagaatcc gatgagaaaa gactaatatt
     4141 tatctatcca ttcagtttga tttctcagga ctttacatcg tcctgaaagt atttgttggt
     4201 accgaggacg cgtaaacatt tacagttgca tgtggcctat tgtttttagc cgttaaatat
     4261 tttataacta ttaaatagcg atacaaattg ttcgaaacta atattgttta tatcatatat
     4321 tctcgcatgt tttaaagctt tattaaattg attttttgta aacagttttt cgtactcttt
     4381 gttaacccat ttcattacaa aagtttcata tttttttctc tctttaaatg ccatttttgc
     4441 tggctttctt tttaatacaa ttaatgtgct atccacttta ggttttggat ggaaataata
     4501 cctaggaatt tttgctaata tagaaatatc tacctctgcc attaacagca atgctagtga
     4561 tctgtttgta tctaataaca ttttagcaaa accatattcc actattaaat aacttattgt
     4621 ggctgaactt tcaaaaacaa tttttcgaat tatatttgtg cttatgttgt aaggtatgct
     4681 gccaaatatt ttatatggat tgtggctagg aaatgtaaat ttcagtatat catcatttac
     4741 tatttgatag ttaggataat ttaagagctt attacgagtt acctcacata atttagaatc
     4801 aatttctatc gccgttacaa aattacatct ctttaccaat ccagcagtaa aatgaccttt
     4861 ccctgcacct atttcaaaga tgttatcttt ttcatctaaa cttatgcaat tcattatttt
     4921 ttctatgtga tattttgaag taataaaatt ttgactatct tttatattta ctttgttcat
     4981 tataacctct ccttaattta ttgcatctct tttcgaatat ttatgttttt tgagaaaaga
     5041 acgtactcat ggttcatccc gatatgcgta tcggtctgta tatcagcaac tttctatgtg
     5101 tttcaactac aatagtcatc tattctcatc tttctgagtc caccccctgc aaagcccctc
     5161 tttacgacat aaaaattcgg tcggaaaagg tatgcaaaag atgtttctct ctttaagaga
     5221 aactcttcgg gatgcaaaaa tatgaaaata actccaattc accaaattat atagcgactt
     5281 ttttacaaaa tgctaaaatt tgttgatttc cgtcaagcaa ttgttgagca aaaatgtctt
     5341 ttacgataaa atgatacctc aatatcaact gtttagcaaa acgatatttc tcttaaagag
     5401 agaaacacct ttttgttcac caatccccga cttttaatcc cgcggccatg attgaaaaag
     5461 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5521 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5581 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5641 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5701 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5761 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5821 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5881 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5941 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     6001 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     6061 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6121 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6181 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6241 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6301 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat aacgcgt
//
