LOCUS       sequence-434372-        8402 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  aTC-inducible expression of secretory BT3630 SP-NanoLuc via 
            P1TDP-A21 promoter-RBS pair in Bacteroides species..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8402)
  AUTHORS   Yeh YH, Kelly VW, Rahman Pour R, Sirk SJ
  TITLE     A molecular toolkit for heterologous protein secretion across 
            Bacteroides species.
  JOURNAL   Nat Commun. 2024 Nov 11;15(1):9741. doi: 10.1038/s41467-024-53845-7.
  PUBMED    39528443
REFERENCE   2  (bases 1 to 8402)
  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: "Nat 
            Commun."; date: "2024-11-11"; volume: "15(1):9741. doi"; pages: " 
            10.1038/s41467-024-53845-7"
COMMENT     Sequence Label: pYHY3
FEATURES             Location/Qualifiers
     source          1..8402
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1028..1651)
                     /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     1559..1578
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    2041..2059
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2103..2121
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             2282..2791
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     CDS             2813..2878
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      2905..2976
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(3083..3874,3875..3943))
                     /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     3706..3725
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3944..4048)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4151..4739
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4640..4659
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            4899..5008
                     /note="incP origin of transfer"
     CDS             5041..5412
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
ORIGIN
        1 taaaagatag tcaaaatttt attacttcaa aatatcacat agaaaaaata atgaattgca
       61 taagtttaga tgaaaaagat aacatctttg aaataggtgc agggaaaggt cattttactg
      121 ctggattggt aaagagatgt aattttgtaa cggcgataga aattgattct aaattatgtg
      181 aggtaactcg taataagctc ttaaattatc ctaactatca aatagtaaat gatgatatac
      241 tgaaatttac atttcctagc cacaatccat ataaaatatt tggcagcata ccttacaaca
      301 taagcacaaa tataattcga aaaattgttt ttgaaagttc agccacaata agttatttaa
      361 tagtggaata tggttttgct aaaatgttat tagatacaaa cagatcacta gcattgctgt
      421 taatggcaga ggtagatatt tctatattag caaaaattcc taggtattat ttccatccaa
      481 aacctaaagt ggatagcaca ttaattgtat taaaaagaaa gccagcaaaa atggcattta
      541 aagagagaaa aaaatatgaa acttttgtaa tgaaatgggt taacaaagag tacgaaaaac
      601 tgtttacaaa aaatcaattt aataaagctt taaaacatgc gagaatatat gatataaaca
      661 atattagttt cgaacaattt gtatcgctat ttaatagtta taaaatattt aacggctaaa
      721 aacaataggc cacatgcaac tgtaaatgtt tacgcgtcct cggtaccaac aaatactttc
      781 aggacgatgt aaagtcctga aaaatcaaag ctggccaatg agacgacggg gtcatcacgg
      841 ctcatcatgc gccaaacaaa tgtgtgcaat acacgctcgg atgactgcat gatgaccgca
      901 ctgactgggg acagcagatc cacctaagcc tgtgagagaa gcagacaccc gacagatcaa
      961 ggcagttaac tagtgcactg cagtggcctt aacggctgac atgggaattc ccctccaccg
     1021 cggtggctta agacccactt tcacatttaa gttgtttttc taatccgcat atgatcaatt
     1081 caaggccgaa taagaaggct ggctctgcac cttggtgatc aaataattcg atagcttgtc
     1141 gtaataatgg cggcatacta tcagtagtag gtgtttccct ttcttcttta gcgacttgat
     1201 gctcttgatc ttccaatacg caacctaaag taaaatgccc cacagcgctg agtgcatata
     1261 atgcattctc tagtgaaaaa ccttgttggc ataaaaaggc taattgattt tcgagagttt
     1321 catactgttt ttctgtaggc cgtgtaccta aatgtacttt tgctccatcg cgatgactta
     1381 gtaaagcaca tctaaaactt ttagcgttat tacgtaaaaa atcttgccag ctttcccctt
     1441 ctaaagggca aaagtgagta tggtgcctat ctaacatctc aatggctaag gcgtcgagca
     1501 aagcccgctt attttttaca tgccaataca atgtaggctg ctctacacct agcttctggg
     1561 cgagtttacg ggttgttaaa ccttcgattc cgacctcatt aagcagctct aatgcgctgt
     1621 taatcacttt acttttatct aatctagaca ttctttaaaa acagatttgg agtgcaaagt
     1681 tacgacaaat aatttgttaa catacatatt taggcaactt tttcactata ttttcaacta
     1741 ataaaacaaa aaagaatgcc gacaagttgt gtgacggcat cctttttcta tttcattagc
     1801 ttatcctaca atcttcttac cttaatttat aatttattcc tgagtcagat caactgcaaa
     1861 gtatccgcgt ttaccggatg ggaatacacc tgtcaggaac aatacctgat tcggagactt
     1921 aacagcagct ttcattgctt cttccagatc atttttgcac ccgctttcca agagaagaaa
     1981 gccttgttaa attgacttag tgtaaaagcg cagtactgct tgaccataag aacaaaaaaa
     2041 tctctatcac tgatagggat aaagtttgga agataaagct aaaagttctt atctttgcag
     2101 tctccctatc agtgatagag acgcatttta aaataaaata aattatttat gatattaaac
     2161 gaatatgatg aaaaagggga tattatttgt gctgacagca gcatttctgg catcttgcca
     2221 gcaggaggaa aacgagggag tagcatctgt ggaccgggtg accatcactc ctattatcac
     2281 agtcttcaca ctcgaagatt tcgttgggga ctggcgacag acagccggct acaacctgga
     2341 ccaagtcctt gaacagggag gtgtgtccag tttgtttcag aatctcgggg tgtccgtaac
     2401 tccgatccaa aggattgtcc tgagcggtga aaatgggctg aagatcgaca tccatgtcat
     2461 catcccgtat gaaggtctga gcggcgacca aatgggccag atcgaaaaaa tttttaaggt
     2521 ggtgtaccct gtggatgatc atcactttaa ggtgatcctg cactatggca cactggtaat
     2581 cgacggggtt acgccgaaca tgatcgacta tttcggacgg ccgtatgaag gcatcgccgt
     2641 gttcgacggc aaaaagatca ctgtaacagg gaccctgtgg aacggcaaca aaattatcga
     2701 cgagcgcctg atcaaccccg acggctccct gctgttccga gtaaccatca acggagtgac
     2761 cggctggcgg ctgtgcgaac gcattctggc gggatcttct ggctcgagcg gcgactacaa
     2821 agaccatgac ggtgattata aagatcatga catcgattac aaggatgacg atgacaagtg
     2881 aatagatcac cattaaccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
     2941 tttcgtttta tctgttgttt gtcggtgaac gctctctaca gcggccgcga ttatcaaaaa
     3001 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3061 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3121 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     3181 gggagggctt accatctggc cccagtgctg caatgatacc gcgggaccca cgctcaccgg
     3241 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3301 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3361 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     3421 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3481 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     3541 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     3601 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     3661 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     3721 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     3781 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     3841 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     3901 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     3961 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4021 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgtcatga
     4081 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4141 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4201 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4261 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     4321 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4381 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4441 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4501 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4561 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4621 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4681 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4741 gcgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     4801 tctttcctgc gttatcccct gattctgtgg ataaccgtgc ggccgcccct ccaaagggtt
     4861 cgtgtagact ttccttggtg tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc
     4921 ccacccgcga gcgggtgttc cttcttcact gtcccttatt cgcacctggc ggtgctcaac
     4981 gggaatcctg ctctgcgagg ctggccggct accgccggcg taacagatga gggcaagcgg
     5041 atggctgatg aaaccaagcc aaccaggaag ggcagcccac ctatcaaggt gtactgcctt
     5101 ccagacgaac gaagagcgat tgaggaaaag gcggcggcgg ccggcatgag cctgtcggcc
     5161 tacctgctgg ccgtcggcca gggctacaaa atcacgggcg tcgtggacta tgagcacgtc
     5221 cgcgagctgg cccgcatcaa tggcgacctg ggccgcctgg gcggcctgct gaaactctgg
     5281 ctcaccgacg acccgcgcac ggcgcggttc ggtgatgcca cgatcctcgc cctgctggcg
     5341 aagatcgaag agaagcagga cgagcttggc aaggtcatga tgggcgtggt ccgcccgagg
     5401 gcagagccat gactttttta gccgctaaaa cggccggggg gtgcgcgtga ttgccaagca
     5461 cgtccccata acgcatagcg ctccccgacc gatgattttt aagtgactga ttttgtgctg
     5521 ttttgggggt atattaagaa taaaagaaat agaataagtt aagtacttga tacacaatat
     5581 agggcatttt ccatattgga aattctcatt ttccaatcta gaaaataccg atttcctaat
     5641 atgatactaa ataggaaaat aatatttccc ttaatattgt ttttatggaa aataatagtt
     5701 tactttgtgg agaataatat ttcccaaaaa catatcaaaa tggaaaataa aaaagcagtt
     5761 aagttaaccg attttcaaaa gaacgaagaa aatcctttta tgaaacaagc tatagaaggt
     5821 attgaaaatc atgttgttaa aaagtataag agtaatagtg gtggcgataa gagagctgta
     5881 gtagctttag ccgacactga aactggagaa gtgtttaaga cttcgtttat ccgtcaaata
     5941 gaagtagatg aagaacaatt cactaaattg tatctttcta actttgctgc attctttgac
     6001 ctatcacaag cagctattcg ggtttttggt tactttatga cctgcatgaa acccaaaaat
     6061 gatttaatca ttttcaatag aaaaaaatgc ctagaatata ccaaatacaa aacagacaaa
     6121 gccgtttata aaggacttgc agaacttgta aaagctgaaa tcatagcccg aggaccagcc
     6181 gataatcttt ggtttattaa tcctctgata gtattcaatg gtgaccgagt gacatttgct
     6241 aaaacatacg ttcggaaaaa gactttagct gcccaaaaga aagaagaagc agagaaacga
     6301 caattatcac ttggctttga tgaacagtaa cactccattg agtgaagctg ccgtttggtc
     6361 gctccccctt tgggggcggg gggggataga taaagttcct ctatgtaaag ttataatggg
     6421 ggatgaaagg caaggttcgc taaccttacc ccgattgttt atgcttccca gcattcaatc
     6481 tatccccgtt tatgggtttc tctttcaccc tacaaagata acctcatggg ggaaaaatgt
     6541 ccaagaatat ggggtaaaaa ctatcaagtc ggtagaaaat aagtatcttt gaagtacatt
     6601 tttagtagag gtactggtgt gcctaggcga agcagatagg caaaaataaa gccgctggag
     6661 atgtcggatg tccgtcaatt aactccagcg gcttgtattt tggtaactct cccgtatcgg
     6721 tcagctaccg tttggtcagc aacaaaggta gtactttatt atcgcaaatc caaatgtttt
     6781 tccctttatt tgtgcattta caaaacattc actaacttcg cacctatgca gtatggtgat
     6841 agcacactag caccctaaag agtgcgtgtg taaggggaca agccccctta accccctgtc
     6901 agccggctgt caccggctgt tatcaatctt caaagtcggg tggttgtcaa gatgattttt
     6961 ttggtttggt gtcgtctttt tttaagctgc cgcataacgg tttggcgccg attgtcacaa
     7021 aaatgttaac ttatgggagc aacaagtatt catgtacaag cagtgaagcc ggggagcgag
     7081 attcacaact ttagggaaaa agagttggac tatgttcgtc ccgaacttag tcatttgaat
     7141 gaaagctggg ttggagatag catttcccat cggctggaga gtgcaaaaca aagatatctc
     7201 gatacggttg ggcagaagat gcaggctaaa gccgcaccca tacgagaggg agtaatagta
     7261 atcaaacaag aaaccactat gcaggaactc cagcagtttg ccacggtctc caaagaacgt
     7321 ttcggtatcg aagcgtttca aatccatata cacaaggacg aaggatacat gaacgcaaag
     7381 cagtggacac ctaaccttca tgcccatgta gttttcgatt ggacgcagcc gaatgggaaa
     7441 agtgtgcgtt tatcgcgtga tgacatggca gaactccaaa ccatagcatc tgaaacactg
     7501 ggcatggaac gtggcgtttc ttctgaccgc aaacacttat cggctatgca gtacaagacc
     7561 gaatgtgcga aagaacagct tcaggaacta tccaacgata tatccagtgc attagacaaa
     7621 tacaaggacg cacaaaacca gcttcttcag cttcagaagg aactacgttc cattgaaaca
     7681 aagaagaacg tccaaaaact catttctaag gcttcagaga agttttacgg cttgattgga
     7741 aaaacagtca acgataggga aaaggatgcc ttaaaagcca aagtaaagac tttagagggg
     7801 gaaaatgaac aactatccga tagactggga aaggctatac ttgaaaaaga gcgaaacagc
     7861 acaaaggcat tcaaagccga gaatgacaag gagtattata gacaacaaat ggacaacgca
     7921 aggacaacca gtaacctatt aaggacggag aaccaaaaac taaaagccga aaccaaggag
     7981 ttgaaaaaag aacttggtaa gatgaaagac ttgttcaatt ccgaacaact ggaggcttta
     8041 aggcatcatt tcccaaacat atcaaaagct atggaagagg ggaaagacct attcaagcaa
     8101 atcaccagaa gcagaggttt cggcatgggt atgtaacccc cccccgcccc caaaggggga
     8161 gcgaccaaac ggcagcctct ctcaatggag tgttacatcc cgtaaagagg ggctttgcag
     8221 ggggtggact cagaaagatg agaatagatg actattgtag ttgaaacaca tagaaagttg
     8281 ctgatataca gaccgatacg catatcggga tgaaccatga gtacgttctt ttctcaaaaa
     8341 acataaatat tcgaaaagag atgcaataaa ttaaggagag gttataatga acaaagtaaa
     8401 ta
//
