LOCUS       sequence-317373-        6195 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Selection by Inducible Effectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6195)
  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 6195)
  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: pSIE1
FEATURES             Location/Qualifiers
     source          1..6195
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(30..821,822..890))
                     /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     653..672
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(2161..2784)
                     /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     2692..2711
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2937..2978
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    3074..3092
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3136..3154
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4208..4226
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4270..4288
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(5377..5393)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5377..5393)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     rep_origin      5426..5814
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            5985..6094
                     /note="incP origin of transfer"
ORIGIN
        1 acgcgttatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
       61 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
      121 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
      181 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
      241 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
      301 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
      361 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
      421 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
      481 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
      541 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
      601 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
      661 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
      721 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
      781 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
      841 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
      901 tttcaatcat ggccgcggga ttaaaagtcg gggattggtg aacaaaaagg tgtttctctc
      961 tttaagagaa atatcgtttt gctaaacagt tgatattgag gtatcatttt atcgtaaaag
     1021 acatttttgc tcaacaattg cttgacggaa atcaacaaat tttagcattt tgtaaaaaag
     1081 tcgctatata atttggtgaa ttggagttat tttcatattt ttgcatcccg aagagtttct
     1141 cttaaagaga gaaacatctt ttgcatacct tttccgaccg aatttttatg tcgtaaagag
     1201 gggctttgca gggggtggac tcagaaagat gagaatagat gactattgta gttgaaacac
     1261 atagaaagtt gctgatatac agaccgatac gcatatcggg atgaaccatg agtacgttct
     1321 tttctcaaaa aacataaata ttcgaaaaga gatgcaataa attaaggaga ggttataatg
     1381 aacaaagtaa atataaaaga tagtcaaaat tttattactt caaaatatca catagaaaaa
     1441 ataatgaatt gcataagttt agatgaaaaa gataacatct ttgaaatagg tgcagggaaa
     1501 ggtcatttta ctgctggatt ggtaaagaga tgtaattttg taacggcgat agaaattgat
     1561 tctaaattat gtgaggtaac tcgtaataag ctcttaaatt atcctaacta tcaaatagta
     1621 aatgatgata tactgaaatt tacatttcct agccacaatc catataaaat atttggcagc
     1681 ataccttaca acataagcac aaatataatt cgaaaaattg tttttgaaag ttcagccaca
     1741 ataagttatt taatagtgga atatggtttt gctaaaatgt tattagatac aaacagatca
     1801 ctagcattgc tgttaatggc agaggtagat atttctatat tagcaaaaat tcctaggtat
     1861 tatttccatc caaaacctaa agtggatagc acattaattg tattaaaaag aaagccagca
     1921 aaaatggcat ttaaagagag aaaaaaatat gaaacttttg taatgaaatg ggttaacaaa
     1981 gagtacgaaa aactgtttac aaaaaatcaa tttaataaag ctttaaaaca tgcgagaata
     2041 tatgatataa acaatattag tttcgaacaa tttgtatcgc tatttaatag ttataaaata
     2101 tttaacggct aaaaacaata ggccacatgc aactgtaaat gtttacgcgt cctcggtacc
     2161 ttaagaccca ctttcacatt taagttgttt ttctaatccg catatgatca attcaaggcc
     2221 gaataagaag gctggctctg caccttggtg atcaaataat tcgatagctt gtcgtaataa
     2281 tggcggcata ctatcagtag taggtgtttc cctttcttct ttagcgactt gatgctcttg
     2341 atcttccaat acgcaaccta aagtaaaatg ccccacagcg ctgagtgcat ataatgcatt
     2401 ctctagtgaa aaaccttgtt ggcataaaaa ggctaattga ttttcgagag tttcatactg
     2461 tttttctgta ggccgtgtac ctaaatgtac ttttgctcca tcgcgatgac ttagtaaagc
     2521 acatctaaaa cttttagcgt tattacgtaa aaaatcttgc cagctttccc cttctaaagg
     2581 gcaaaagtga gtatggtgcc tatctaacat ctcaatggct aaggcgtcga gcaaagcccg
     2641 cttatttttt acatgccaat acaatgtagg ctgctctaca cctagcttct gggcgagttt
     2701 acgggttgtt aaaccttcga ttccgacctc attaagcagc tctaatgcgc tgttaatcac
     2761 tttactttta tctaatctag acataatgtt attttttaaa tgttgtgtga tcagtcctac
     2821 tttgtttctt tcgacactgc aaatataaga acattatttg aaagttcaag tgaaacttta
     2881 aattttaaca atagattaac cattgcaaac aaaacaaaaa aaaggtagcc caattgtaaa
     2941 acgaaaggcc cagtctttcg actgagcctt tcgttttagt cgactttttg cacccgcttt
     3001 ccaagagaag aaagccttgt taaattgact tagtgtaaaa gcgcagtact gcttgaccat
     3061 aagaacaaaa aaatctctat cactgatagg gataaagttt ggaagataaa gctaaaagtt
     3121 cttatctttg cagtctccct atcagtgata gagacgaaat aaagacatat aaaagaaaag
     3181 acaccatgaa gaaaatttta tctttgcttg tgatggctat tgtagctata cagttctcat
     3241 ttgccggcgg taaggaaata gaaataaaga aattgccaga gttcgaggct atggtgaatg
     3301 ccggtaatac gacctatacg ggattggtag aaggcatcgg gtatgtgtat tggacaacag
     3361 aaaccctata ttttatccgc actaatccag aacaattatg ggcgattcca aaatatcagc
     3421 aaataccttt cccctatttt caaaggaaag atgcaatcat tgagaccaaa acattacata
     3481 cactccatgt cttgtcaaaa gatgaactgt tgaaattgga ttacgatgcc tattatgcat
     3541 tttatggtat cgtggaggag atgctaaaat ttattcatcg ggcggatgct attaaaagtt
     3601 attgtgaaat acctcttccc ataataaaat ccaaaggagc actaaaggga aatgacgcaa
     3661 gaaatgggat actatcatta ggtagtcaaa taaatgacca aatcggagct ccccttgatg
     3721 cagcgaatat gctaatggat aataaacata taggcaaaat tggagatgga ttgtcgctta
     3781 tttctattat agatgaggta ggtaatggtg aatattggtc agctgccgga gacattttac
     3841 tgtttgcagc cggaaaaaca aaattaagtc cctatatgac tgtcataagt ttaggcacat
     3901 ggatgtatga gacggacttg atgcaatgga gattagcatg tataaattat agcgattaca
     3961 aaaaaacact aataaaatat cgagaattac aaaaaaaatt tgaaagtgga gacaaatctg
     4021 tagaggaaaa gatgaatgaa tgtcacaaaa tactgaattc acattatata gagatgcaaa
     4081 aaaatttagg taatctagga gttaaattct aactgcagtt tttgcacccg ctttccaaga
     4141 gaagaaagcc ttgttaaatt gacttagtgt aaaagcgcag tactgcttga ccataagaac
     4201 aaaaaaatct ctatcactga tagggataaa gtttggaaga taaagctaaa agttcttatc
     4261 tttgcagtct ccctatcagt gatagagacg aaataaagac atataaaaga aaagacacca
     4321 tgaagaaaat tttatctttg cttgtgatgg ctattgtagc tatacagttc tcatttgccg
     4381 gcgcttatag aaatgaatat aaaacgcaaa gtgaatttaa tgtcatcttg gaacgggggg
     4441 atgactatga aggatttgta gttggcttgg gatatacttg gatgagtagc aaggtaattc
     4501 taccggtaaa tcaaaacgga tggagtccaa tatctcggaa tgtttcggtg gacgagagtt
     4561 ttcatacgat agtctcagaa aggaaatacg atacctccca atatgcctat gaaaaaagtc
     4621 tgatgcaaga tccaacaaaa gtttccgaaa aggtccgtga cttaatagtt aaaaacaaag
     4681 gtaataatat cactgagata aatttaggcc aagaaaagca atatttgccc acggataata
     4741 gtcagataag tattgtaatt actgacactg gcagtcggta cgaaattgtg attagtgcca
     4801 cggataactc aaatggaaag acttatgagg caaagtatga gagtttgacg gacttggtgt
     4861 cagcggtacg cgattctggt agcctgcctg ctgtaaataa ggaaggaccc aatctggaag
     4921 gactggcagg gttaggattc ggaattgctg aaacagccgg aaattgggct gagaagatta
     4981 tggataatcg aggtgcgtac ttacctaagc agatgcgttt ctcgcccaaa acgcttccgc
     5041 cgattataag attgccttta gggaactatc aagtccccgc taaaggtatg agtagagtgc
     5101 gtggagtagg taaagctttg ggatgggcag gaatggtgct tactggctat caagttgtgc
     5161 gtgatgtgca aaatggtcga tttgccgtgg caggtacaag aattgccgta gcaggtttag
     5221 cttatggcgt tacttttatt ccctatgtcg gttgggtctt agctatcggt atcggagtgg
     5281 ctgattatac ttggggtgac gagttctatg actggataga caatagagct tctgaattgg
     5341 aaatgtggtg ggacggtgta agattagcat tatgaggatc cactagttct agagcggccg
     5401 ccaccgcggt ggaggggaat tcccatgtca gccgttaagt gttcctgtgt cactcaaaat
     5461 tgctttgaga ggctctaagg gcttctcagt gcgttacatc cctggcttgt tgtccacaac
     5521 cgttaaacct taaaagcttt aaaagcctta tatattcttt tttttcttat aaaacttaaa
     5581 accttagagg ctatttaagt tgctgattta tattaatttt attgttcaaa catgagagct
     5641 tagtacgtga aacatgagag cttagtacgt tagccatgag agcttagtac gttagccatg
     5701 agggtttagt tcgttaaaca tgagagctta gtacgttaaa catgagagct tagtacgtga
     5761 aacatgagag cttagtacgt actatcaaca ggttgaactg ctgatcttca gatcctctac
     5821 gccggacgca tcgtggccgg atcaattccg ttttccgctg cataaccctg cttcggggtc
     5881 attatagcga ttttttcggt atatccatcc tttttcgcac gatatacagg attttgccaa
     5941 agggttcgtg tagactttcc ttggtgtatc caacggcgtc agccgggcag gataggtgaa
     6001 gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca cctggcggtg
     6061 ctcaacggga atcctgctct gcgaggctgg ccggctaccg ccggcgtaac agatgagggc
     6121 aagcggatgg ctgatgaaac caagccaacc aggaagggca gcccacctat cacggaattg
     6181 atccccctcg aattg
//
