LOCUS       sequence-496782-        7615 bp    DNA     circular SYN 09-JAN-2026
DEFINITION  Transcriptional reporter backbone/entry vector with mGL and 
            constitutive mScarlet-I3 in pNBU2 in pNBU2 integrative Bacteroides 
            plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7615)
  TITLE     Tropini Lab plasmids
REFERENCE   2  (bases 1 to 7615)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 9, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pEM526
FEATURES             Location/Qualifiers
     source          1..7615
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             207..893
                     /codon_start=1
                     /product="bright cysteine-free monomeric red fluorescent 
                     protein with fast and complete maturation (Gadella et al., 
                     2023)"
                     /label=mScarlet3
                     /translation="MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYWKQSFPEGFKWERVMIFEDGGT
                     VSVTQDTSLEDGTLIYKVKLRGGNFPPDGPVMQKRTMGWEASTERLYPEDVVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHS
                     TGGSGGS"
     primer_bind     complement(337..355)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      919..946
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             1018..1641
                     /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     complement(1091..1110)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    1806..1824
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             join(1911..1913,1914..1916,1917..2630)
                     /codon_start=1
                     /product="monomeric GFP derivative that displays 
                     exceptionally high brightness in cells (Campbell et al., 
                     2020)"
                     /label=mGreenLantern
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLGYGVACFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIVLKGIDFKEDGNILGHKLEYNFNSHKVYITADKQKNGIK
                     ANFKTRHNVEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLK
                     ERVTAAGITHDMDELYK"
     primer_bind     complement(1956..1977)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     terminator      2651..2678
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(join(4658..5449,5450..5518))
                     /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     5281..5300
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6810..7198
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            7369..7478
                     /note="incP origin of transfer"
ORIGIN
        1 caattgggct accttttttt tgttttgttt gcaatggtta atctattgtt aaaatttaaa
       61 gtttcacttg aactttcaaa taatgttctt atatttgcag tgtcgaaaga aacaaagtag
      121 gactgatcac acaacattta aaaaataaca ttatgaaagc actcgaaaag agattcaaag
      181 gtaatattaa caataattta ttttcaatgg atagcaccga ggcagtgatc aaggagttca
      241 tgcggttcaa ggtgcacatg gagggctcca tgaacggcca cgagttcgag atcgagggcg
      301 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
      361 gccccctgcc cttctcctgg gacatcctgt cccctcagtt catgtacggc tccagggcct
      421 tcatcaagca ccccgccgac atccccgact actggaagca gtccttcccc gagggcttca
      481 agtgggagcg cgtgatgatc ttcgaggacg gcggcaccgt gtccgtgacc caggacacct
      541 ccctggagga cggcaccctg atctacaagg tgaagctccg cggcggcaac ttccctcctg
      601 acggccccgt aatgcagaag aggacaatgg gctgggaagc atccaccgag cggttgtacc
      661 ccgaggacgt cgtgctgaag ggcgacatta agatggccct gcgcctgaag gacggcggcc
      721 gctacctggc ggacttcaag accacctaca aggccaagaa gcccgtgcag atgcccggcg
      781 ccttcaacat cgaccgcaag ttggacatca cctcccacaa cgaggactac accgtggtgg
      841 aacagtacga acgctccgtg gcccgccact ccaccggcgg ctccggtggc tcctaataaa
      901 tcctagctga ttagctcgag aaggccatcc tgacggatgg cctttttttt gtatggggac
      961 tctgagaacc ctacctttca aaccccgaag agaccgcgga tgtacgaagt ggtctcgatg
     1021 tccagattag ataaaagtaa agtgattaac agcgcattag agctgcttaa tgaggtcgga
     1081 atcgaaggtt taacaacccg taaactcgcc cagaagctag gtgtagagca gcctacattg
     1141 tattggcatg taaaaaataa gcgggctttg ctcgacgcct tagccattga gatgttagat
     1201 aggcaccata ctcacttttg ccctttagaa ggggaaagct ggcaagattt tttacgtaat
     1261 aacgctaaaa gttttagatg tgctttacta agtcatcgcg atggagcaaa agtacattta
     1321 ggtacacggc ctacagaaaa acagtatgaa actctcgaaa atcaattagc ctttttatgc
     1381 caacaaggtt tttcactaga gaatgcatta tatgcactca gcgctgtggg gcattttact
     1441 ttaggttgcg tattggaaga tcaagagcat caagtcgcta aagaagaaag ggaaacacct
     1501 actactgata gtatgccgcc attattacga caagctatcg aattatttga tcaccaaggt
     1561 gcagagccag ccttcttatt cggccttgaa ttgatcatat gcggattaga aaaacaactt
     1621 aaatgtgaaa gtgggtctta atggccacat cggttcctgc tagaaggccc atcctgacgg
     1681 atggcctttt ggtattcgag cattacctca tactttggat ttttgttttg tttgcaatgg
     1741 ttaatctatt gttaaaattt aaagtttcac ttgaactttc aaataatgtt cttatatttg
     1801 cagtgtccct atcagtgata gagagtagga tcacacaaca tttaaaaaat aacattatga
     1861 aagcactcga aaagagattc aaaggtaata ttaacaataa tttattttca atggtgagca
     1921 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1981 acggccacaa gttcagcgtc cgcggcgagg gcgagggcga tgccaccaac ggcaagctga
     2041 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     2101 ccttaggcta cggcgtggcc tgcttcgccc gctaccccga ccacatgaag cagcacgact
     2161 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatctct ttcaaggacg
     2221 acggtaccta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     2281 tcgtgctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     2341 acaacttcaa cagccacaag gtctatatca cggccgacaa gcagaagaac ggcatcaagg
     2401 ctaacttcaa gacccgccac aacgttgagg acggcggcgt gcagctcgcc gaccactacc
     2461 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagcc
     2521 atcagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgaaggaga
     2581 gggtgaccgc cgccgggatt acacatgaca tggacgagct gtacaagtaa tagcgttcag
     2641 ctcgactgtg agaaggccat cctgacggat ggcctttttt tttcgttata actagttgcg
     2701 ttcctagccg ctactagtgt aattgcctat cttccagtga tggaacagca tttgtgcatt
     2761 ggctgcaaca atcagcctta cttgtgcctg ttctatttcc gaaccgaccg cttgtatgaa
     2821 tgcatcaaaa ttcgttttct ctacgttgga ttccttgttg ctcatattgt gatgataatt
     2881 tctacaaata tagtcattgg taactatcta tgaaactgtt tgatactttt atagttgatt
     2941 aaacttgttc atggcatttg ccttaatatc atccgctatg tcaatgtagg gtttcatagc
     3001 tttgtagtcg ctgtgtcccg tccatttcat gaccacctgt gccgggattc cgagagccag
     3061 cgcattgcag atgaatgtcc tttttcctgc atgggtactg agcaaagcgt atttgtgtgt
     3121 gacttcatca atacgttcat ttcccttgta gtaggtttcc cgtacaggct cgttgatttc
     3181 tgccagttcg cccagctctt tcaggtaatc gttcatcttc tggttgctga tgacgggcag
     3241 agccatgtaa ttctcgaaat ggatgtcctt gtatttgtcc agtatggctt tgctgtattt
     3301 gttcagttca atcgtcaggc tgtcggcagt cttgactgtg gttatttcga tgtggtcgga
     3361 cttcacatcg cttcttttca gattgcgaac atccgaatac cgcaaactcg taaagcagca
     3421 gaacaggaaa acatcacgca cacgttccag gtattgctta tccttgggta tctggtagtc
     3481 tttcagcttg ttcagttcat cccaagtcag gaagattact tttttcgagg tggttttcag
     3541 tttcggtttg aacgtatcgt atgcaatgtt ctgatgatgt cctttcttga agctccagcg
     3601 caggaaccat ttgaggaatc ccatttgctt gccgatggtg ctgtttctca tatccttggt
     3661 gtcacgcagg aagttgacgt attcgttcaa tccaaactcg ttgaaatagt tgaacgttgc
     3721 atcctccttg aactctttga ggtggttcct cactgctgca aatttttcat aggtggatgc
     3781 cgtccagtta ttctggttac cgcactcttt tacaaactca tcgaacacct cccaaaagct
     3841 gacaggggct tcttccggct gttcttcact ggtgtctttc attctcatgt tgaaagcttc
     3901 cttcaactgt tgggtcgttg gcatgacctc ctgcacctca aattccttga aaatattctg
     3961 gatttcggca tagtatttca gcaagtccgt attgatttcg gctgcacttt gctttagctt
     4021 gttggtacat ccgttcttta cccgctgctt atctgcatcc catttggcta cgtcaatccg
     4081 gtagcccgtt gtaaactcga tacgttggct ggcaaagatg acacgcatac ggatgggtac
     4141 gttctctacg attggcacac cgttcttttt ccggctctcc aatgcaaaaa tgatgttgcg
     4201 cttgatattc ataattgggt gcgtttgaaa ttctacaccc aaatatacac ccaattattg
     4261 agatagcaaa agacatttag aaacatttac ttttactcta tattgtaatt tacacttgat
     4321 tatcagtcgt ttgcagtttt atgatattct gtgaaagtat aagttcgaga gcctgtctct
     4381 ccgcaaaaaa cgctgaaaat cagcagattg caaaacaaac accctgtttt acacccaaga
     4441 atgtaaagtc gggtgttttt gttttattta agataataca accactacat aataaaagag
     4501 tagcgatatt aaaagaatcc gatgagaaaa gactaatatt tatctatcca ttcagtttga
     4561 tttctcagga ctttacatcg tcctgaaagt atttgttggt accggtaccg aggacgcgac
     4621 agctgcagac gcgttatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4681 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4741 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcggg
     4801 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4861 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4921 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4981 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5041 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5101 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5161 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5221 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5281 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5341 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5401 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5461 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5521 tcttcctttt tcaatcatgg ccgcgggatt aaaagtcggg gattggtgaa caaaaaggtg
     5581 tttctctctt taagagaaat atcgttttgc taaacagttg atattgaggt atcattttat
     5641 cgtaaaagac atttttgctc aacaattgct tgacggaaat caacaaattt tagcattttg
     5701 taaaaaagtc gctatataat ttggtgaatt ggagttattt tcatattttt gcatcccgaa
     5761 gagtttctct taaagagaga aacatctttt gcataccttt tccgaccgaa tttttatgtc
     5821 gtaaagaggg gctttgcagg gggtggactc agaaagatga gaatagatga ctattgtagt
     5881 tgaaacacat agaaagttgc tgatatacag accgatacgc atatcgggat gaaccatgag
     5941 tacgttcttt tctcaaaaaa cataaatatt cgaaaagaga tgcaataaat taaggagagg
     6001 ttataatgaa caaagtaaat ataaaagata gtcaaaattt tattacttca aaatatcaca
     6061 tagaaaaaat aatgaattgc ataagtttag atgaaaaaga taacatcttt gaaataggtg
     6121 cagggaaagg tcattttact gctggattgg taaagagatg taattttgta acggcgatag
     6181 aaattgattc taaattatgt gaggtaactc gtaataagct cttaaattat cctaactatc
     6241 aaatagtaaa tgatgatata ctgaaattta catttcctag ccacaatcca tataaaatat
     6301 ttggcaacat accttacaac ataagcacaa atataattcg aaaaattgtt tttgaaagtt
     6361 cagccacaat aagttattta atagtggaat atggttttgc taaaatgtta ttagatacaa
     6421 acagatcact agcattgctg ttaatggcag aggtagatat ttctatatta gcaaaaattc
     6481 ctaggtatta tttccatcca aaacctaaag tggatagcac attaattgta ttaaaaagaa
     6541 agccagcaaa aatggcattt aaagagagaa aaaaatatga aacttttgta atgaaatggg
     6601 ttaacaaaga gtacgaaaaa ctgtttacaa aaaatcaatt taataaagct ttaaaacatg
     6661 cgagaatata tgatataaac aatattagtt tcgaacaatt tgtatcgcta tttaatagtt
     6721 ataaaatatt taacggctaa aaacaatagg ccacatgcaa ctgtaaatgt ttacgcgggt
     6781 accgacaccg cggtggaggg gaattcccat gtcagccgtt aagtgttcct gtgtcactca
     6841 aaattgcttt gagaggctct aagggcttct cagtgcgtta catccctggc ttgttgtcca
     6901 caaccgttaa accttaaaag ctttaaaagc cttatatatt cttttttttc ttataaaact
     6961 taaaacctta gaggctattt aagttgctga tttatattaa ttttattgtt caaacatgag
     7021 agcttagtac gtgaaacatg agagcttagt acgttagcca tgagagctta gtacgttagc
     7081 catgagggtt tagttcgtta aacatgagag cttagtacgt taaacatgag agcttagtac
     7141 gtgaaacatg agagcttagt acgtactatc aacaggttga actgctgatc ttcagatcct
     7201 ctacgccgga cgcatcgtgg ccggatcaat tccgttttcc gctgcataac cctgcttcgg
     7261 ggtcattata gcgatttttt cggtatatcc atcctttttc gcacgatata caggattttg
     7321 ccaaagggtt cgtgtagact ttccttggtg tatccaacgg cgtcagccgg gcaggatagg
     7381 tgaagtaggc ccacccgcga gcgggtgttc cttcttcact gtcccttatt cgcacctggc
     7441 ggtgctcaac gggaatcctg ctctgcgagg ctggccggct accgccggcg taacagatga
     7501 gggcaagcgg atggctgatg aaaccaagcc aaccaggaag ggcagcccac ctatcacgga
     7561 attgatcccc ctcgaattgg aattctagcc tggaataggg aagcaccaca tcacc
//
