LOCUS       sequence-210293-        5793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AraCAM + AraE + PBAD-YFP Reporter. Ara sensor for use in 
            non-Marionette strain..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5793)
  AUTHORS   Meyer AJ, Segall-Shapiro TH, Glassey E, Zhang J, Voigt CA
  TITLE     Escherichia coli ""Marionette"" strains with 12 highly optimized 
            small-molecule sensors.
  JOURNAL   Nat Chem Biol. 2018 Nov 26. pii: 10.1038/s41589-018-0168-3. doi: 
            10.1038/s41589-018-0168-3.
  PUBMED    30478458
REFERENCE   2  (bases 1 to 5793)
  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 Chem 
            Biol. 2018 Nov 26. pii: 10.1038/s41589-018-0168-3. doi: 
            10.1038/s41589-018-0168-3."
COMMENT     Sequence Label: pAJM.677
FEATURES             Location/Qualifiers
     source          1..5793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        1..50
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             join(100..102,103..105,106..819)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(145..166)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(406..425)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     753..774
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             967..1845
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDFFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MLAINGSLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGLE
                     EKVNDVAVKLS"
     primer_bind     complement(3497..3516)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      complement(3881..4426)
                     /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(3961..3980)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4515..5330)
                     /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(4515..5330)
                     /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     5238..5257
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5322..5341
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        5503..5786
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     5709..5728
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 ctgtcaccgg atgtgctttc cggtctgatg agtccgtgag gacgaaacag cctctacaaa
       61 taattttgtt taatactaga gaaagagggg aaatactaga tggtgagcaa gggcgaggag
      121 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
      181 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
      241 atctgcacca caggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac
      301 ggcctgcaat gcttcgcccg ctaccccgac cacatgaagc tgcacgactt cttcaagtcc
      361 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
      421 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
      481 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
      541 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
      601 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
      661 ccaatcggcg acggccccgt gctgctgccc gacaaccact accttagcta ccagtccgcc
      721 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
      781 gccgggatca ctctcggcat ggacgagctg tacaagtaac tcggtaccaa attccagaaa
      841 agaggcctcc cgaaaggggg gccttttttc gttttggtcc aatggcggcg cgccatcgaa
      901 tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
      961 gtgaatatgg ctgaagcgca aaatgatccc ctgctgccgg gatactcgtt taatgcccat
     1021 ctggtggcgg gtttaacgcc gattgaggcc aacggttatc tcgatttttt tatcgaccga
     1081 ccgctgggaa tgaaaggtta tattctcaat ctcaccattc gcggtcaggg ggtggtgaaa
     1141 aatcagggac gagaatttgt ttgccgaccg ggtgatattt tgctgttccc gccaggagag
     1201 attcatcact acggtcgtca tccggaggct cgcgaatggt atcaccagtg ggtttacttt
     1261 cgtccgcgcg cctactggca tgaatggctt aactggccgt caatatttgc caatacgggg
     1321 ttctttcgcc cggatgaagc gcaccagccg catttcagcg acttttttgg gcaaatcatt
     1381 aacgccgggc aaggggaagg gcgctattcg gagctgctgg cgataaatct gcttgagcaa
     1441 ttgttactgc ggcgcatgct agcgattaac ggatcgctcc atccaccgat ggataatcgg
     1501 gtacgcgagg cttgtcagta catcagcgat cacctggcag acagcaattt tgatatcgcc
     1561 agcgtcgcac agcatgtttg cttgtcgccg tcgcgtctgt cacatctttt ccgccagcag
     1621 ttagggatta gcgtcttaag ctggcgcgag gaccaacgta tcagccaggc gaagctgctt
     1681 ttgagcacca cccggatgcc tatcgccacc gtcggtcgca atgttggttt tgacgatcaa
     1741 ctctatttct cgcgggtatt taaaaaatgc accggggcca gcccgagcga gttccgtgcc
     1801 ggtttggaag aaaaagtgaa tgatgtagcc gtcaagttgt catgataaga tcctattcca
     1861 gcgggattaa agaggagcga ttaagcatgg ttactatcaa tacggaatct gctttaacgc
     1921 cacgttcttt gcgggatacg cggcgtatga atatgtttgt ttcggtagct gctgcggtcg
     1981 caggattgtt atttggtctt gatatcggcg taatcgccgg agcgttgccg ttcattaccg
     2041 atcactttgt gctgaccagt cgtttgcagg aatgggtggt tagtagcatg atgctcggtg
     2101 cagcaattgg tgcgctgttt aatggttggc tgtcgttccg cctggggcgt aaatacagcc
     2161 tgatggcggg ggccatcctg tttgtactcg gttctatagg gtccgctttt gcgaccagcg
     2221 tagagatgtt aatcgccgct cgtgtggtgc tgggcattgc tgtcgggatc gcgtcttaca
     2281 ccgctcctct gtatctttct gaaatggcaa gtgaaaacgt tcgcggtaag atgatcagta
     2341 tgtaccagtt gatggtcaca ctcggcatcg tgctggcgtt tttatccgat acagcgttca
     2401 gttatagcgg taactggcgc gcaatgttgg gggttcttgc tttaccagca gttctgctga
     2461 ttattctggt agtatttctg ccaaatagcc cgcgctggct ggcggaaaag gggcgtcata
     2521 ttgaggcgga agaagtattg cgtatgctgc gcgatacgtc ggaaaaagcg cgagaagaac
     2581 tcaacgaaat tcgtgaaagc ctgaagttaa aacagggcgg ttgggcactg tttaagatca
     2641 accgtaacgt ccgtcgtgct gtgtttctcg gtatgttgtt gcaggcgatg cagcagttta
     2701 ccggtatgaa catcatcatg tactacgcgc cgcgtatctt caaaatggcg ggctttacga
     2761 ccacagaaca acagatgatt gcgactctgg tcgtagggct gacctttatg ttcgccacct
     2821 ttattgcggt gtttacggta gataaagcag ggcgtaaacc ggctctgaaa attggtttca
     2881 gcgtgatggc gttaggcact ctggtgctgg gctattgcct gatgcagttt gataacggta
     2941 cggcttccag tggcttgtcc tggctctctg ttggcatgac gatgatgtgt attgccggtt
     3001 atgcgatgag cgccgcgcca gtggtgtgga tcctgtgctc tgaaattcag ccgctgaaat
     3061 gccgcgattt cggtattacc tgttcgacca ccacgaactg ggtgtcgaat atgattatcg
     3121 gcgcgacctt cctgacactg cttgatagca ttggcgctgc cggtacgttc tggctctaca
     3181 ctgcgctgaa cattgcgttt gtgggcatta ctttctggct cattccggaa accaaaaatg
     3241 tcacgctgga acatatcgaa cgcaaactga tggcaggcga gaagttgaga aatatcggcg
     3301 tctgataagg atcctaattg gtaacgaatc agacaattga cggctcgagg gagtagcata
     3361 gggtttgcag aatccctgct tcgtccattt gacaggcaca ttatgcatcg atgataagct
     3421 gtcaaacatg agcagatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca
     3481 ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac
     3541 ttcgggctca tgagcaaata ttttatctga ggtgcttcct cgctcactga ctcgctgcac
     3601 gaggcagacc tcagcgctag cggagtgtat actggcttac tatgttggca ctgatgaggg
     3661 tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg caccggtgcg tcagcagaat
     3721 atgtgataca ggatatattc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga
     3781 ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt tcctggaaga tgccaggaag
     3841 atacttaaca gggaagtgag agggccgcgg caaagccgtt tttccatagg ctccgccccc
     3901 ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat
     3961 aaagatacca ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc
     4021 ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc
     4081 agttccgggt aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc
     4141 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa
     4201 gcaccactgg cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc
     4261 cggttaaggc taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc
     4321 tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt
     4381 ttcgttttca gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat
     4441 taaggggtct gacgctcagt ggaacgaaaa atcaatctaa agtatatatg agtaaacttg
     4501 gtctgacagt taccttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata
     4561 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
     4621 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
     4681 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
     4741 ccatgagtga cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact
     4801 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
     4861 ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta
     4921 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     4981 cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg
     5041 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     5101 tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg
     5161 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca
     5221 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     5281 gaatttaatc gcggcctgga gcaagacgtt tcccgttgaa tatggctcat aacacccctt
     5341 gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt
     5401 gcaatgtaca tcagagattt tgagacacaa ccaattattg aaggcctccc taacgggggg
     5461 cctttttttg tttctggtct cccgcttaac gatcgttggc tgagaaacca attgtccata
     5521 ttgcatcaga cattgccgtc actgcgtctt ttactggctc ttctcgctaa ccaaaccggt
     5581 aaccccgctt attaaaagca ttctgtaaca aagcgggacc aaagccatga caaaaacgcg
     5641 taacaaaagt gtctataatc acggcagaaa agtccacatt gattatttgc acggcgtcac
     5701 actttgctat gccatagcat ttttatccat aagattagcg gatcctacct gacgcttttt
     5761 atcgcaactc tctactgttt ctccataccc gag
//
