LOCUS       sequence-368890-        6804 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The pET15 PemrR-gfp biosensor is transcriptional fusion of GFP with 
            the promoter of the emrRAB operon. This biosensor is responsive to 
            monoaromatics, such as vanillin and syringaldehyde..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6804)
  AUTHORS   Ho JCH, Pawar SV, Hallam SJ, Yadav VG
  TITLE     An Improved Whole-Cell Biosensor for the Discovery of 
            Lignin-Transforming Enzymes in Functional Metagenomic Screens.
  JOURNAL   ACS Synth Biol. 2018 Feb 16;7(2):392-398. doi: 
            10.1021/acssynbio.7b00412. Epub 2017 Dec 4.
  PUBMED    29182267
REFERENCE   2  (bases 1 to 6804)
  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: 
            "10.1021/acssynbio.7b00412"; journalName: "ACS Synth Biol"; date: 
            "2018-02-16- 16"; volume: "7"; issue: "2"; pages: "392-398"
COMMENT     Sequence Label: pET15 PemrR-gfp biosensor
FEATURES             Location/Qualifiers
     source          1..6804
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(981..2063)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     2025..2044
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2064..2141)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2347..2366
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             2669..3382
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2699..2727)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3322..3342
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3402..3473
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3489..3516
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     3549..3568
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3549..3567
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3568..3592
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             3607..3629
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3672..3692
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3754..3772)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3768..3815
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3990..3995,3996..4012,4013..4018))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(4045..4063)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4131..4235
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4236..4304,4305..5096)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4454..4473)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5267..5855
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5756..5775
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6009..6026
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6041..6180
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6106..6125)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6266..6288
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6282..6473)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca
       61 ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg
      121 atcatgcgca cccgtggcca ggacccaacg ctgcccgaga tgcgccgcgt gcggctgctg
      181 gagatggcgg acgcgatgga tatgttctgc caagggttgg tttgcgcatt cacagttctc
      241 cgcaagaatt gattggctcc aattcttgga gtggtgaatc cgttagcgag gtgccgccgg
      301 cttccattca ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga
      361 caaggtatag ggcggcgcct acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg
      421 cataaatcgc cgtgacgatc agcggtccaa tgatcgaagt taggctggta agagccgcga
      481 gcgatccttg aagctgtccc tgatggtcgt catctacctg cctggacagc atggcctgca
      541 acgcgggcat cccgatgccg ccggaagcga gaagaatcat aatggggaag gccatccagc
      601 ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa
      661 tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg
      721 cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc
      781 ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa
      841 agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga
      901 ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac
      961 ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     1021 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg
     1081 gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga
     1141 gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg
     1201 gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag
     1261 atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc
     1321 tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt
     1381 tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga
     1441 ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat
     1501 gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt
     1561 cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca
     1621 agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc
     1681 agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct
     1741 ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga
     1801 tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag
     1861 gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga
     1921 atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg
     1981 ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca
     2041 tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat
     2101 catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc
     2161 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc
     2221 cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
     2281 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     2341 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggcgactag
     2401 tcgcagcatt atcatcccaa cactgcttag tgcgctggcc tatgggctcg cctggaaagt
     2461 gatggcgatt atataaccca caagaatcat ttttctaaaa caatacattt actttatttg
     2521 tcactgtcgt tactatatcg gctgaaatta atgaggtcat acccaaatgg atagttcgtt
     2581 tacgcccatt gaacaaatgc taaaatttcg cgccagccgc cacgaagatt ttccttggat
     2641 cgtctagatt taagaaggag atatagatat gagtaaagga gaagaacttt tcactggagt
     2701 tgtcccaatt cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtcagtgg
     2761 agagggtgaa ggtgatgcaa catacggaaa acttaccctt aaatttattt gcactactgg
     2821 aaaactacct gttccatggc caacacttgt cactactttc gcgtatggtc ttcaatgctt
     2881 tgcgagatac ccagatcaca tgaaacagca tgactttttc aagagtgcca tgcccgaagg
     2941 ttatgtacag gaaagaacta tatttttcaa agatgacggg aactacaaga cacgtgctga
     3001 agtcaagttt gaaggtgata cccttgttaa tagaatcgag ttaaaaggta ttgattttaa
     3061 agaagatgga aacattcttg gacacaaatt ggaatacaac tataactcac acaatgtata
     3121 catcatggca gacaaacaaa agaatggaat caaagttaac ttcaaaatta gacacaacat
     3181 tgaagatgga agcgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg
     3241 ccctgtcctt ttaccagaca accattacct gtccacacaa tctgcccttt cgaaagatcc
     3301 caacgaaaag agagaccaca tggtccttct tgagtttgta acagctgctg ggattacaca
     3361 tggcatggat gaattgtaca aataatacta gagccaggca tcaaataaaa cgaaaggctc
     3421 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga
     3481 gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta tagctagcag atctcgatcc
     3541 cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga
     3601 aataattttg tttaacttta agaaggagat ataccatggg cagcagccat catcatcatc
     3661 acagcagcgg cgaaaacctg tattttcagg gccatatgct cgaggatccg gctgctaaca
     3721 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
     3781 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat
     3841 atcccgcaag aggcccggca gtaccggcat aaccaagcct atgcctacag catccagggt
     3901 gacggtgccg aggatgacga tgagcgcatt gttagatttc atacacggtg cctgactgcg
     3961 ttagcaattt aactgtgata aactaccgca ttaaagctta tcgatgataa gctgtcaaac
     4021 atgagaattc ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     4081 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4141 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4201 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     4261 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     4321 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     4381 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     4441 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac
     4501 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     4561 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     4621 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     4681 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4741 aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc
     4801 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4861 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4921 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     4981 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     5041 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     5101 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5161 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5221 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5281 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5341 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5401 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5461 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5521 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5581 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5641 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5701 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5761 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5821 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5881 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5941 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     6001 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     6061 ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga
     6121 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     6181 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     6241 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     6301 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     6361 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     6421 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
     6481 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
     6541 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
     6601 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc
     6661 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
     6721 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa
     6781 ccgaagacca ttcatgttgt tgct
//
