LOCUS       sequence-378047-        6887 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Propionate sensor expressing eYFP reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6887)
  AUTHORS   Lebovich M, Andrews LB
  TITLE     Surveying the Genetic Design Space for Transcription Factor-Based 
            Metabolite Biosensors: Synthetic Gamma-Aminobutyric Acid and 
            Propionate Biosensors in E. coli Nissle 1917.
  JOURNAL   Front Bioeng Biotechnol. 2022 Aug 25;10:938056. doi: 
            10.3389/fbioe.2022.938056. eCollection 2022.
  PUBMED    36091463
REFERENCE   2  (bases 1 to 6887)
  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: "Front 
            Bioeng Biotechnol."; date: "2022-08-25"; volume: "10:938056. doi"; 
            pages: " 10.3389/fbioe.2022.938056. eCollection 2022"
COMMENT     Sequence Label: pML2003
FEATURES             Location/Qualifiers
     source          1..6887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(75..94)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(2128..2145)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2262..2807)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2342..2361)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2896..3711)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2896..3711)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3619..3638
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3703..3722
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     misc_RNA        4084..4133
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             join(4183..4185,4186..4188,4189..4902)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4228..4249)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4489..4508)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4836..4857
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5055..6137
                     /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     complement(5074..5093)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             6156..6779
                     /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(6229..6248)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
ORIGIN
        1 caaacatgag cagatcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg
       61 tgcggttgct ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt
      121 cgggctcatg agcaaatatt ttatctgagg tgcttcctcg ctcacagagg ataagtccct
      181 aactttgaca atgcctagat caccgtatat aatgctagca ctgacgagac tacgagcggt
      241 caacgcatgt gctttgcgtt ctgatgagac agtgatgtcg aaaccgcctc tacaaataat
      301 tttgtttaac ccaatattcg gtttcttaac tttgcggtgc gctatggcac atccaccacg
      361 gcttaatgac gacaaaccgg ttatctggac ggtatctgta acgcgcctgt tcgagctgtt
      421 tcgcgatatc agcctcgagt ttgatcacct ggcgaacatt acccctatcc agcttggctt
      481 tgaaaaagcg gtgacctaca tccacaagaa actggcaaac gaacgctgtg acgccatcat
      541 cgccgctgga tctaacggcg cgtacctgaa aagccgcctg tcagtgccag ttattttgat
      601 taaaccgagc ggctacgatg tgttacaggc actggcaaaa gccggaaaac tcacctcttc
      661 tatcggcgtt gtcacttatc aggaaactat tccggcactg gtggcgtttc aaaaaacctt
      721 taatttgcgc ctcgatcaac gtagctacat taccgaagaa gatgcacgcg ggcagattaa
      781 cgagctaaaa gctaacggca ccgaagcggt ggtcggcgcg gggctgatta ccgatctggc
      841 agaagaagcc ggaatgaccg gaatttttat ctattccgcc gccaccgtgc gccaggcgtt
      901 cagcgatgcg ctggatatga cgcgcatgtc gttacgccat aacactcacg atgccacccg
      961 caacgccctg agaactcgtt acgtgctggg cgatatgctc ggtcaatcac cacagatgga
     1021 acaagtacgg cagactattt tgctgtatgc ccgctccagt gcagcggtgt tgattgaggg
     1081 ggaaacgggg acgggcaaag agctggcggc ccaggcgatt catcgggaat attttgcccg
     1141 ccacgatgcg cgacagggca aaaagtcgca tccgtttgtt gcagtcaact gcggggcgat
     1201 tgccgaatcg ctgctggaag cagaactgtt tggctatgag gaaggggcgt ttaccggctc
     1261 gcgacgcggc ggtcgcgccg ggctgtttga aatcgcccac ggaggtacgc tgtttctcga
     1321 tgagattggc gaaatgccgc tgccgttgca gacccggctg ctgcgggtgc tggaagaaaa
     1381 agaggtcacc cgcgtcggcg ggcatcagcc tgttccggtg gatgtgcggg tcattagcgc
     1441 cactcactgc aatctggaag aagatatgcg gcaagggcag tttcgccgtg acctgtttta
     1501 tcggctgagt attttgcgtc tgcaattgcc accactgcgc gagcgggtgg cggatattct
     1561 gccactggcg gaaagctttt tgaaagtgtc tctggcggcg ctctccaccc cgttttctgc
     1621 cgcattacgc cagggattac aggcaagcga aaccgtgctg gtgcactacg actggccggg
     1681 caatattcgt gaactgcgca atatgatgga gcgactggcg ctatttttaa gtgtggaacc
     1741 gacgccggat ttaacgccgc aatttttgca gctgctactg ccggaactgg cgcgcgagtc
     1801 ggcgaaaact cccgctccac gcttactgac accacaacag gcactggaga aatttaatgg
     1861 cgataaaaca gcagcggcga attatttagg catcagccgg acgacgttct ggcggcggct
     1921 gaaaagctga gtgaaaaaaa aaaaacaccc taacgggtgt tttttttttt ttggtctccc
     1981 atcggcagac ctcagcgcta gcggagtgta tactagctta ctatgttggc actgatgagg
     2041 gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa
     2101 tatgtgatac aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg
     2161 actgcggcga gcggaaatgg cttacgaacg gggcggagat ttcccggaag atgccaggaa
     2221 gatacttaac agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc
     2281 cctgacaagc atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta
     2341 taaagatacc aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt tcctgccttt
     2401 cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact
     2461 cagttccggg taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc
     2521 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa
     2581 agcaccactg gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg
     2641 ccggttaagg ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac
     2701 ctcggttcaa agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt
     2761 tttcgttttc agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta
     2821 ttaaggggtc tgacgctcag tggaacgaaa aatcaatcta aagtatatat gagtaaactt
     2881 ggtctgacag ttaccttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     2941 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     3001 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     3061 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     3121 accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac
     3181 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     3241 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
     3301 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     3361 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     3421 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     3481 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     3541 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     3601 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     3661 ggaatttaat cgcggcctgg agcaagacgt ttcccgttga atatggctca taacacccct
     3721 tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg
     3781 tgcaatgtac atcagagatt ttgagacaca accaattatt gaaggcctcc ctaacggggg
     3841 gccttttttt gtttctggtc tcccgcttgt cttgtttcat aattgttgca atgaaacgcg
     3901 gtgaaacatt gcctgaaacg ttaactgaaa cgcatatttg cggattagtt catgacttta
     3961 tctctaacaa attgaaatta aacatttaat tttattaagg caattgtggc acaccccttg
     4021 ctttgtcttt atcaacgcaa ataacaagtt gataacaagc tagcaggagg aattcacctg
     4081 aagctgtcac cggatgtgct ttccggtctg atgagtccgt gaggacgaaa cagcctctac
     4141 aaataatttt gtttaatact agagaaagag gggaaatact agatggtgag caagggcgag
     4201 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     4261 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     4321 ttcatctgca ccacaggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcggc
     4381 tacggcctgc aatgcttcgc ccgctacccc gaccacatga agctgcacga cttcttcaag
     4441 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     4501 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     4561 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     4621 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     4681 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     4741 accccaatcg gcgacggccc cgtgctgctg cccgacaacc actaccttag ctaccagtcc
     4801 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     4861 gccgccggga tcactctcgg catggacgag ctgtacaagt aactcggtac caaattccag
     4921 aaaagaggcc tcccgaaagg ggggcctttt ttcgttttgg tccaaatgaa tggcggcgcg
     4981 ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat
     5041 tcagggtggt gaatatgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct
     5101 cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcgtg
     5161 aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac
     5221 tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc
     5281 cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg
     5341 tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg
     5401 cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg
     5461 tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca
     5521 tcaacagtat tattttctcc catgaggacg gtacgcgact gggcgtggag catctggtcg
     5581 cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc
     5641 tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac
     5701 gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg
     5761 gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg
     5821 ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata
     5881 ccgaagatag ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc
     5941 tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca
     6001 atcagctgtt gccagtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa
     6061 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     6121 tggaaagcgg gcagtgacaa tccaggagga aaaaaatgtc cagattagat aaaagtaaag
     6181 tgattaacag cgcattagag ctgcttaatg aggtcggaat cgaaggttta acaacccgta
     6241 aactcgccca gaagctaggt gtagagcagc ctacattgta ttggcatgta aaaaataagc
     6301 gggctttgct cgacgcctta gccattgaga tgttagatag gcaccatact cacttttgcc
     6361 ctttagaagg ggaaagctgg caagattttt tacgtaataa cgctaaaagt tttagatgtg
     6421 ctttactaag tcatcgcgat ggagcaaaag tacatttagg tacacggcct acagaaaaac
     6481 agtatgaaac tctcgaaaat caattagcct ttttatgcca acaaggtttt tcactagaga
     6541 atgcattata tgcactcagc gctgtggggc attttacttt aggttgcgta ttggaagatc
     6601 aagagcatca agtcgctaaa gaagaaaggg aaacacctac tactgatagt atgccgccat
     6661 tattacgaca agctatcgaa ttatttgatc accaaggtgc agagccagcc ttcttattcg
     6721 gccttgaatt gatcatatgc ggattagaaa aacaacttaa atgtgaaagt gggtcctaat
     6781 aattggtaac gaatcagaca attgacggct cgagggagta gcatagggtt tgcagaatcc
     6841 ctgcttcgtc catttgacag gcacattatg catcgatgat aagctgt
//
