LOCUS       sequence-270200-        3126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PBAD-YFP Reporter.  Ara sensor for use in Marionette strain..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3126)
  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 3126)
  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.716
FEATURES             Location/Qualifiers
     source          1..3126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     302..321
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     561..582
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_RNA        complement(677..726)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     promoter        complement(734..1017)
                     /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     complement(792..811)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(1179..1198)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1190..2005
                     /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             1190..2005
                     /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     complement(1263..1282)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2094..2639
                     /direction=RIGHT
                     /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     2540..2559
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3034..3126,1..621,622..624,625..627))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3079..3100)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gctcagggcg gactggtagc taaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc
       61 gattggggtg ttctgctggt agtggtcggc gagctgcacg ctgccgtcct cgatgttgtg
      121 gcggatcttg aagttcacct tgatgccgtt cttctgcttg tcggccatga tatagacgtt
      181 gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc
      241 gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg
      301 ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt agccttcggg
      361 catggcggac ttgaagaagt cgtgcagctt catgtggtcg gggtagcggg cgaagcattg
      421 caggccgtag ccgaaggtgg tcacgagggt gggccagggc acgggcagct tgcctgtggt
      481 gcagatgaac ttcagggtca gcttgccgta ggtggcatcg ccctcgccct cgccggacac
      541 gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt
      601 gaacagctcc tcgcccttgc tcaccatcta gtatttcccc tctttctcta gtattaaaca
      661 aaattatttg tagaggctgt ttcgtcctca cggactcatc agaccggaaa gcacatccgg
      721 tgacagctcg ggtatggaga aacagtagag agttgcgata aaaagcgtca ggtaggatcc
      781 gctaatctta tggataaaaa tgctatggca tagcaaagtg tgacgccgtg caaataatca
      841 atgtggactt ttctgccgtg attatagaca cttttgttac gcgtttttgt catggctttg
      901 gtcccgcttt gttacagaat gcttttaata agcggggtta ccggtttggt tagcgagaag
      961 agccagtaaa agacgcagtg acggcaatgt ctgatgcaat atggacaatt ggtttctcag
     1021 ccaacgatcg ttaagcggga gaccagaaac aaaaaaaggc cccccgttag ggaggccttc
     1081 aataattggt tgtgtctcaa aatctctgat gtacattgca caagataaaa atatatcatc
     1141 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     1201 tcaacgggaa acgtcttgct ccaggccgcg attaaattcc aacatggatg ctgatttata
     1261 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgattgta
     1321 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     1381 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     1441 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccgggaa
     1501 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     1561 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     1621 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     1681 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     1741 gcttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     1801 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     1861 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     1921 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     1981 tttgatgctc gatgagtttt tctaaggtaa ctgtcagacc aagtttactc atatatactt
     2041 tagattgatt tttcgttcca ctgagcgtca gaccccttaa taagatgatc ttcttgagat
     2101 cgttttggtc tgcgcgtaat ctcttgctct gaaaacgaaa aaaccgcctt gcagggcggt
     2161 ttttcgaagg ttctctgagc taccaactct ttgaaccgag gtaactggct tggaggagcg
     2221 cagtcaccaa aacttgtcct ttcagtttag ccttaaccgg cgcatgactt caagactaac
     2281 tcctctaaat caattaccag tggctgctgc cagtggtgct tttgcatgtc tttccgggtt
     2341 ggactcaaga cgatagttac cggataaggc gcagcggtcg gactgaacgg ggggttcgtg
     2401 catacagtcc agcttggagc gaactgccta cccggaactg agtgtcaggc gtggaatgag
     2461 acaaacgcgg ccataacagc ggaatgacac cggtaaaccg aaaggcagga acaggagagc
     2521 gcacgaggga gccgccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     2581 accactgatt tgagcgtcag atttcgtgat gcttgtcagg ggggcggagc ctatggaaaa
     2641 acggctttgc cgcggccctc tcacttccct gttaagtatc ttcctggcat cttccaggaa
     2701 atctccgccc cgttcgtaag ccatttccgc tcgccgcagt cgaacgaccg agcgtagcga
     2761 gtcagtgagc gaggaagcgg aatatatcct gtatcacata ttctgctgac gcaccggtgc
     2821 agcctttttt ctcctgccac atgaagcact tcactgacac cctcatcagt gccaacatag
     2881 taagccagta tacactccgc tagcgctgag gtctgcctcg tgcagcgagt cagtgagcga
     2941 ggaagcacct cattcgatgg cgcgccgcca ttggaccaaa acgaaaaaag gccccccttt
     3001 cgggaggcct cttttctgga atttggtacc gagttacttg tacagctcgt ccatgccgag
     3061 agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg
     3121 gtcttt
//
