LOCUS       sequence-313687-        8087 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Selection by Inducible Effectors Test Plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8087)
  AUTHORS   Bencivenga-Barry NA, Lim B, Herrera CM, Trent MS, Goodman AL
  TITLE     Genetic Manipulation of Wild Human Gut Bacteroides.
  JOURNAL   J Bacteriol. 2020 Jan 15;202(3). pii: JB.00544-19. doi: 
            10.1128/JB.00544-19. Print 2020 Jan 15.
  PUBMED    31712278
REFERENCE   2  (bases 1 to 8087)
  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"; journalName: "J 
            Bacteriol."; date: "2020-01-15"; volume: "202(3). pii: JB.00544-19. 
            doi"; pages: " 10.1128/JB.00544-19. Print 2020 Jan 15"
COMMENT     Sequence Label: pNBU2_erm_SIE1
FEATURES             Location/Qualifiers
     source          1..8087
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    42..60
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    104..122
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1176..1194
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1238..1256
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             join(5530..5598,5599..6390)
                     /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(5748..5767)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            6521..6630
                     /note="incP origin of transfer"
     rep_origin      6801..7189
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(7216..7839)
                     /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     7747..7766
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      7992..8033
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gtgtaaaagc gcagtactgc ttgaccataa gaacaaaaaa atctctatca ctgataggga
       61 taaagtttgg aagataaagc taaaagttct tatctttgca gtctccctat cagtgataga
      121 gacgaaataa agacatataa aagaaaagac accatgaaga aaattttatc tttgcttgtg
      181 atggctattg tagctataca gttctcattt gccggcggta aggaaataga aataaagaaa
      241 ttgccagagt tcgaggctat ggtgaatgcc ggtaatacga cctatacggg attggtagaa
      301 ggcatcgggt atgtgtattg gacaacagaa accctatatt ttatccgcac taatccagaa
      361 caattatggg cgattccaaa atatcagcaa atacctttcc cctattttca aaggaaagat
      421 gcaatcattg agaccaaaac attacataca ctccatgtct tgtcaaaaga tgaactgttg
      481 aaattggatt acgatgccta ttatgcattt tatggtatcg tggaggagat gctaaaattt
      541 attcatcggg cggatgctat taaaagttat tgtgaaatac ctcttcccat aataaaatcc
      601 aaaggagcac taaagggaaa tgacgcaaga aatgggatac tatcattagg tagtcaaata
      661 aatgaccaaa tcggagctcc ccttgatgca gcgaatatgc taatggataa taaacatata
      721 ggcaaaattg gagatggatt gtcgcttatt tctattatag atgaggtagg taatggtgaa
      781 tattggtcag ctgccggaga cattttactg tttgcagccg gaaaaacaaa attaagtccc
      841 tatatgactg tcataagttt aggcacatgg atgtatgaga cggacttgat gcaatggaga
      901 ttagcatgta taaattatag cgattacaaa aaaacactaa taaaatatcg agaattacaa
      961 aaaaaatttg aaagtggaga caaatctgta gaggaaaaga tgaatgaatg tcacaaaata
     1021 ctgaattcac attatataga gatgcaaaaa aatttaggta atctaggagt taaattctaa
     1081 ctgcagtttt tgcacccgct ttccaagaga agaaagcctt gttaaattga cttagtgtaa
     1141 aagcgcagta ctgcttgacc ataagaacaa aaaaatctct atcactgata gggataaagt
     1201 ttggaagata aagctaaaag ttcttatctt tgcagtctcc ctatcagtga tagagacgaa
     1261 ataaagacat ataaaagaaa agacaccatg aagaaaattt tatctttgct tgtgatggct
     1321 attgtagcta tacagttctc atttgccggc gcttatagaa atgaatataa aacgcaaagt
     1381 gaatttaatg tcatcttgga acggggggat gactatgaag gatttgtagt tggcttggga
     1441 tatacttgga tgagtagcaa ggtaattcta ccggtaaatc aaaacggatg gagtccaata
     1501 tctcggaatg tttcggtgga cgagagtttt catacgatag tctcagaaag gaaatacgat
     1561 acctcccaat atgcctatga aaaaagtctg atgcaagatc caacaaaagt ttccgaaaag
     1621 gtccgtgact taatagttaa aaacaaaggt aataatatca ctgagataaa tttaggccaa
     1681 gaaaagcaat atttgcccac ggataatagt cagataagta ttgtaattac tgacactggc
     1741 agtcggtacg aaattgtgat tagtgccacg gataactcaa atggaaagac ttatgaggca
     1801 aagtatgaga gtttgacgga cttggtgtca gcggtacgcg attctggtag cctgcctgct
     1861 gtaaataagg aaggacccaa tctggaagga ctggcagggt taggattcgg aattgctgaa
     1921 acagccggaa attgggctga gaagattatg gataatcgag gtgcgtactt acctaagcag
     1981 atgcgtttct cgcccaaaac gcttccgccg attataagat tgcctttagg gaactatcaa
     2041 gtccccgcta aaggtatgag tagagtgcgt ggagtaggta aagctttggg atgggcagga
     2101 atggtgctta ctggctatca agttgtgcgt gatgtgcaaa atggtcgatt tgccgtggca
     2161 ggtacaagaa ttgccgtagc aggtttagct tatggcgtta cttttattcc ctatgtcggt
     2221 tgggtcttag ctatcggtat cggagtggct gattatactt ggggtgacga gttctatgac
     2281 tggatagaca atagagcttc tgaattggaa atgtggtggg acggtgtaag attagcatta
     2341 tgatgcagga attcgatatc aagcttatcg ataccgtcga ctaattgcct atcttccagt
     2401 gatggaacag catttgtgca ttggctgcaa caatcagcct tacttgtgcc tgttctattt
     2461 ccgaaccgac cgcttgtatg aatgcatcaa aattcgtttt ctctacgttg gattccttgt
     2521 tgctcatatt gtgatgataa tttctacaaa tatagtcatt ggtaactatc tatgaaactg
     2581 tttgatactt ttatagttga ttaaacttgt tcatggcatt tgccttaata tcatccgcta
     2641 tgtcaatgta gggtttcata gctttgtagt cgctgtgtcc cgtccatttc atgaccacct
     2701 gtgccgggat tccgagagcc agcgcattgc agatgaatgt cctttttcct gcatgggtac
     2761 tgagcaaagc gtatttgggt gtgacttcat caatacgttc atttcccttg tagtaggttt
     2821 cccgtacagg ctcgttgatt tctgccagtt cgcccagctc tttcaggtaa tcgttcatct
     2881 tctggttgct gatgacgggc agagccatgt aattctcgaa atggatgtcc ttgtatttgt
     2941 ccagtatggc tttgctgtat ttgttcagtt caatcgtcag gctgtcggca gtcttgactg
     3001 tggttatttc gatgtggtcg gacttcacat cgcttctttt cagattgcga acatccgaat
     3061 accgcaaact cgtaaagcag cagaacagga aaacatcacg cacacgttcc aggtattgct
     3121 tatccttggg tatctggtag tctttcagct tgttcagttc atcccaagtc aggaagatta
     3181 cttttttcga ggtggttttc agtttcggtt tgaacgtatc gtatgcaatg ttctgatgat
     3241 gtcctttctt gaagctccag cgcaggaacc atttgaggaa tcccatttgc ttgccgatgg
     3301 tgctgtttct catatccttg gtgtcacgca ggaagttgac gtattcgttc aatccaaact
     3361 cgttgaaata gttgaacgtt gcatcctcct tgaactcttt gaggtggttc ctcactgctg
     3421 caaatttttc ataggtggat gccgtccagt tattctggtt accgcactct tttacaaact
     3481 catcgaacac ctcccaaaag ctgacagggg cttcttccgg ctgttcttca ctggtgtctt
     3541 tcattctcat gttgaaagct tccttcaact gttgggtcgt tggcatgacc tcctgcacct
     3601 caaattcctt gaaaatattc tggatttcgg catagtattt cagcaagtcc gtattgattt
     3661 cggctgcact ttgctttagc ttgttggtac atccgttctt tacccgctgc ttatctgcat
     3721 cccatttggc tacgtcaatc cggtagcccg ttgtaaactc gatacgttgg ctggcaaaga
     3781 tgacacgcat acggatgggt acgttctcta cgattggcac accgttcttt ttccggctct
     3841 ccaatgcaaa aatgatgttg cgcttgatat tcataattgg gtgcgtttga aattctacac
     3901 ccaaatatac acccaattat tgagatagca aaagacattt agaaacattt acttttactc
     3961 tatattgtaa tttacacttg attatcagtc gtttgcagtt ttatgatatt ctgtgaaagt
     4021 ataagttcga gagcctgtct ctccgcaaaa aacgctgaaa atcagcagat tgcaaaacaa
     4081 acaccctgtt ttacacccaa gaatgtaaag tcgggtgttt ttgttttatt taagataata
     4141 caaccactac ataataaaag agtagcgata ttaaaagaat ccgatgagaa aagactaata
     4201 tttatctatc cattcagttt gatttctcag gactttacat cgtcctgaaa gtatttgttg
     4261 gtaccgagga cgcgtaaaca tttacagttg catgtggcct attgttttta gccgttaaat
     4321 attttataac tattaaatag cgatacaaat tgttcgaaac taatattgtt tatatcatat
     4381 attctcgcat gttttaaagc tttattaaat tgattttttg taaacagttt ttcgtactct
     4441 ttgttaaccc atttcattac aaaagtttca tatttttttc tctctttaaa tgccattttt
     4501 gctggctttc tttttaatac aattaatgtg ctatccactt taggttttgg atggaaataa
     4561 tacctaggaa tttttgctaa tatagaaata tctacctctg ccattaacag caatgctagt
     4621 gatctgtttg tatctaataa cattttagca aaaccatatt ccactattaa ataacttatt
     4681 gtggctgaac tttcaaaaac aatttttcga attatatttg tgcttatgtt gtaaggtatg
     4741 ctgccaaata ttttatatgg attgtggcta ggaaatgtaa atttcagtat atcatcattt
     4801 actatttgat agttaggata atttaagagc ttattacgag ttacctcaca taatttagaa
     4861 tcaatttcta tcgccgttac aaaattacat ctctttacca atccagcagt aaaatgacct
     4921 ttccctgcac ctatttcaaa gatgttatct ttttcatcta aacttatgca attcattatt
     4981 ttttctatgt gatattttga agtaataaaa ttttgactat cttttatatt tactttgttc
     5041 attataacct ctccttaatt tattgcatct cttttcgaat atttatgttt tttgagaaaa
     5101 gaacgtactc atggttcatc ccgatatgcg tatcggtctg tatatcagca actttctatg
     5161 tgtttcaact acaatagtca tctattctca tctttctgag tccaccccct gcaaagcccc
     5221 tctttacgac ataaaaattc ggtcggaaaa ggtatgcaaa agatgtttct ctctttaaga
     5281 gaaactcttc gggatgcaaa aatatgaaaa taactccaat tcaccaaatt atatagcgac
     5341 ttttttacaa aatgctaaaa tttgttgatt tccgtcaagc aattgttgag caaaaatgtc
     5401 ttttacgata aaatgatacc tcaatatcaa ctgtttagca aaacgatatt tctcttaaag
     5461 agagaaacac ctttttgttc accaatcccc gacttttaat cccgcggcca tgattgaaaa
     5521 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     5581 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5641 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5701 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5761 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5821 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5881 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5941 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6001 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6061 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6121 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6181 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6241 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6301 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6361 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc ataacgcgtc
     6421 aattcgaggg ggatcaattc cgtgataggt gggctgccct tcctggttgg cttggtttca
     6481 tcagccatcc gcttgccctc atctgttacg ccggcggtag ccggccagcc tcgcagagca
     6541 ggattcccgt tgagcaccgc caggtgcgaa taagggacag tgaagaagga acacccgctc
     6601 gcgggtgggc ctacttcacc tatcctgccc ggctgacgcc gttggataca ccaaggaaag
     6661 tctacacgaa ccctttggca aaatcctgta tatcgtgcga aaaaggatgg atataccgaa
     6721 aaaatcgcta taatgacccc gaagcagggt tatgcagcgg aaaacggaat tgatccggcc
     6781 acgatgcgtc cggcgtagag gatctgaaga tcagcagttc aacctgttga tagtacgtac
     6841 taagctctca tgtttcacgt actaagctct catgtttaac gtactaagct ctcatgttta
     6901 acgaactaaa ccctcatggc taacgtacta agctctcatg gctaacgtac taagctctca
     6961 tgtttcacgt actaagctct catgtttgaa caataaaatt aatataaatc agcaacttaa
     7021 atagcctcta aggttttaag ttttataaga aaaaaaagaa tatataaggc ttttaaagct
     7081 tttaaggttt aacggttgtg gacaacaagc cagggatgta acgcactgag aagcccttag
     7141 agcctctcaa agcaattttg agtgacacag gaacacttaa cggctgacat gggaattccc
     7201 ctccaccgcg gtggcttaag acccactttc acatttaagt tgtttttcta atccgcatat
     7261 gatcaattca aggccgaata agaaggctgg ctctgcacct tggtgatcaa ataattcgat
     7321 agcttgtcgt aataatggcg gcatactatc agtagtaggt gtttcccttt cttctttagc
     7381 gacttgatgc tcttgatctt ccaatacgca acctaaagta aaatgcccca cagcgctgag
     7441 tgcatataat gcattctcta gtgaaaaacc ttgttggcat aaaaaggcta attgattttc
     7501 gagagtttca tactgttttt ctgtaggccg tgtacctaaa tgtacttttg ctccatcgcg
     7561 atgacttagt aaagcacatc taaaactttt agcgttatta cgtaaaaaat cttgccagct
     7621 ttccccttct aaagggcaaa agtgagtatg gtgcctatct aacatctcaa tggctaaggc
     7681 gtcgagcaaa gcccgcttat tttttacatg ccaatacaat gtaggctgct ctacacctag
     7741 cttctgggcg agtttacggg ttgttaaacc ttcgattccg acctcattaa gcagctctaa
     7801 tgcgctgtta atcactttac ttttatctaa tctagacata atgttatttt ttaaatgttg
     7861 tgtgatcagt cctactttgt ttctttcgac actgcaaata taagaacatt atttgaaagt
     7921 tcaagtgaaa ctttaaattt taacaataga ttaaccattg caaacaaaac aaaaaaaagg
     7981 tagcccaatt gtaaaacgaa aggcccagtc tttcgactga gcctttcgtt ttagtcgact
     8041 ttttgcaccc gctttccaag agaagaaagc cttgttaaat tgactta
//
