LOCUS       sequence-180031-        5896 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration vector at an attB site in Bacteroides; contains 1kb 
            cassette with both the P1T_DP aTC-inducible promoter (RBS GH023) and
            the consitutively-driven TetR by the P2-A21 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5896)
  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 5896)
  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
FEATURES             Location/Qualifiers
     source          1..5896
                     /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             join(5007..5075,5076..5867)
                     /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(5225..5244)
                     /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 ccatggtgga tcccccgggc tgcaggaatt cgatatcaag cttatcgata ccgtcgacta
     1861 attgcctatc ttccagtgat ggaacagcat ttgtgcattg gctgcaacaa tcagccttac
     1921 ttgtgcctgt tctatttccg aaccgaccgc ttgtatgaat gcatcaaaat tcgttttctc
     1981 tacgttggat tccttgttgc tcatattgtg atgataattt ctacaaatat agtcattggt
     2041 aactatctat gaaactgttt gatactttta tagttgatta aacttgttca tggcatttgc
     2101 cttaatatca tccgctatgt caatgtaggg tttcatagct ttgtagtcgc tgtgtcccgt
     2161 ccatttcatg accacctgtg ccgggattcc gagagccagc gcattgcaga tgaatgtcct
     2221 ttttcctgca tgggtactga gcaaagcgta tttgggtgtg acttcatcaa tacgttcatt
     2281 tcccttgtag taggtttccc gtacaggctc gttgatttct gccagttcgc ccagctcttt
     2341 caggtaatcg ttcatcttct ggttgctgat gacgggcaga gccatgtaat tctcgaaatg
     2401 gatgtccttg tatttgtcca gtatggcttt gctgtatttg ttcagttcaa tcgtcaggct
     2461 gtcggcagtc ttgactgtgg ttatttcgat gtggtcggac ttcacatcgc ttcttttcag
     2521 attgcgaaca tccgaatacc gcaaactcgt aaagcagcag aacaggaaaa catcacgcac
     2581 acgttccagg tattgcttat ccttgggtat ctggtagtct ttcagcttgt tcagttcatc
     2641 ccaagtcagg aagattactt ttttcgaggt ggttttcagt ttcggtttga acgtatcgta
     2701 tgcaatgttc tgatgatgtc ctttcttgaa gctccagcgc aggaaccatt tgaggaatcc
     2761 catttgcttg ccgatggtgc tgtttctcat atccttggtg tcacgcagga agttgacgta
     2821 ttcgttcaat ccaaactcgt tgaaatagtt gaacgttgca tcctccttga actctttgag
     2881 gtggttcctc actgctgcaa atttttcata ggtggatgcc gtccagttat tctggttacc
     2941 gcactctttt acaaactcat cgaacacctc ccaaaagctg acaggggctt cttccggctg
     3001 ttcttcactg gtgtctttca ttctcatgtt gaaagcttcc ttcaactgtt gggtcgttgg
     3061 catgacctcc tgcacctcaa attccttgaa aatattctgg atttcggcat agtatttcag
     3121 caagtccgta ttgatttcgg ctgcactttg ctttagcttg ttggtacatc cgttctttac
     3181 ccgctgctta tctgcatccc atttggctac gtcaatccgg tagcccgttg taaactcgat
     3241 acgttggctg gcaaagatga cacgcatacg gatgggtacg ttctctacga ttggcacacc
     3301 gttctttttc cggctctcca atgcaaaaat gatgttgcgc ttgatattca taattgggtg
     3361 cgtttgaaat tctacaccca aatatacacc caattattga gatagcaaaa gacatttaga
     3421 aacatttact tttactctat attgtaattt acacttgatt atcagtcgtt tgcagtttta
     3481 tgatattctg tgaaagtata agttcgagag cctgtctctc cgcaaaaaac gctgaaaatc
     3541 agcagattgc aaaacaaaca ccctgtttta cacccaagaa tgtaaagtcg ggtgtttttg
     3601 ttttatttaa gataatacaa ccactacata ataaaagagt agcgatatta aaagaatccg
     3661 atgagaaaag actaatattt atctatccat tcagtttgat ttctcaggac tttacatcgt
     3721 cctgaaagta tttgttggta ccgaggacgc gtaaacattt acagttgcat gtggcctatt
     3781 gtttttagcc gttaaatatt ttataactat taaatagcga tacaaattgt tcgaaactaa
     3841 tattgtttat atcatatatt ctcgcatgtt ttaaagcttt attaaattga ttttttgtaa
     3901 acagtttttc gtactctttg ttaacccatt tcattacaaa agtttcatat ttttttctct
     3961 ctttaaatgc catttttgct ggctttcttt ttaatacaat taatgtgcta tccactttag
     4021 gttttggatg gaaataatac ctaggaattt ttgctaatat agaaatatct acctctgcca
     4081 ttaacagcaa tgctagtgat ctgtttgtat ctaataacat tttagcaaaa ccatattcca
     4141 ctattaaata acttattgtg gctgaacttt caaaaacaat ttttcgaatt atatttgtgc
     4201 ttatgttgta aggtatgctg ccaaatattt tatatggatt gtggctagga aatgtaaatt
     4261 tcagtatatc atcatttact atttgatagt taggataatt taagagctta ttacgagtta
     4321 cctcacataa tttagaatca atttctatcg ccgttacaaa attacatctc tttaccaatc
     4381 cagcagtaaa atgacctttc cctgcaccta tttcaaagat gttatctttt tcatctaaac
     4441 ttatgcaatt cattattttt tctatgtgat attttgaagt aataaaattt tgactatctt
     4501 ttatatttac tttgttcatt ataacctctc cttaatttat tgcatctctt ttcgaatatt
     4561 tatgtttttt gagaaaagaa cgtactcatg gttcatcccg atatgcgtat cggtctgtat
     4621 atcagcaact ttctatgtgt ttcaactaca atagtcatct attctcatct ttctgagtcc
     4681 accccctgca aagcccctct ttacgacata aaaattcggt cggaaaaggt atgcaaaaga
     4741 tgtttctctc tttaagagaa actcttcggg atgcaaaaat atgaaaataa ctccaattca
     4801 ccaaattata tagcgacttt tttacaaaat gctaaaattt gttgatttcc gtcaagcaat
     4861 tgttgagcaa aaatgtcttt tacgataaaa tgatacctca atatcaactg tttagcaaaa
     4921 cgatatttct cttaaagaga gaaacacctt tttgttcacc aatccccgac ttttaatccc
     4981 gcggccatga ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     5041 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     5101 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     5161 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     5221 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     5281 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     5341 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     5401 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     5461 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     5521 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     5581 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     5641 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5701 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     5761 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     5821 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     5881 tttactcata acgcgt
//
