LOCUS       sequence-503210-        2898 bp    DNA     circular SYN 17-MAR-2026
DEFINITION  RNA toehold switch to detect genomic sequences of Shiga toxin II 
            controlling expression of sfgfp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2898)
  AUTHORS   Zhang Y, Kojima T, Kim GA, McNerney MP, Takayama S, Styczynski MP
  TITLE     Protocell arrays for simultaneous detection of diverse analytes.
  JOURNAL   Nat Commun. 2021 Sep 29;12(1):5724. doi: 10.1038/s41467-021-25989-3.
  PUBMED    34588445
REFERENCE   2  (bases 1 to 2898)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 17, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2021-09-29"; volume: "12(1):5724. doi"; pages: " 
            10.1038/s41467-021-25989-3"
COMMENT     Sequence Label: pStx2Switch_GFP
FEATURES             Location/Qualifiers
     source          1..2898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     617..636
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        617..635
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             736..1449
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(766..794)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1389..1409
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1472..1501
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1517..1534
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1590..1607)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1760..1806
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1883..1901)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1897..1944
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1962..2777)
                     /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="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVYALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2685..2704
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2769..2788
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacat cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcgatcccg cgaaattaat acgactcact atagggggat actatttaac gaagagacat
      661 aactttgttg ggtcggactt tagaacagag gagataaaga tggacccaac aaagaacctg
      721 gcggcagcgc aaaagatgcg taaaggagaa gaacttttca ctggagttgt cccaattctt
      781 gttgaattag atggtgatgt taatgggcac aaattttctg tccgtggaga gggtgaaggt
      841 gatgctacaa acggaaaact cacccttaaa tttatttgca ctactggaaa actacctgtt
      901 ccgtggccaa cacttgtcac tactctgacc tatggtgttc aatgcttttc ccgttatccg
      961 gatcacatga aacggcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa
     1021 cgcactatat ctttcaaaga tgacgggacc tacaagacgc gtgctgaagt caagtttgaa
     1081 ggtgataccc ttgttaatcg tatcgagtta aagggtattg attttaaaga agatggaaac
     1141 attcttggac acaaactcga gtacaacttt aactcacaca atgtatacat cacggcagac
     1201 aaacaaaaga atggaatcaa agctaacttc aaaattcgcc acaacgttga agatggttcc
     1261 gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta
     1321 ccagacaacc attacctgtc gacacaatct gtcctttcga aagatcccaa cgaaaagcgt
     1381 gaccacatgg tccttcttga gtttgtaact gctgctggga ttacacatgg catggatgag
     1441 ctctacaaat aaggatctga agcttgggcc cgaacaaaaa ctcatctcag aagaggatct
     1501 gaatagcgcc gtcgaccatc atcatcatca tcattgagtt taaacggtct ccagcttggc
     1561 tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc
     1621 ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg
     1681 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga
     1741 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     1801 ttttatctgt tgtttgtcgg tgaactggat cgtcgaccgg ctgctaacaa agcccgaaag
     1861 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
     1921 aaacgggtct tgaggggttt tttgctgaaa gccaattctg attagaaaaa ctcatcgagc
     1981 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
     2041 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     2101 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     2161 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     2221 aaaagcttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     2281 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat
     2341 acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
     2401 actgccagcg cataaacaat attttcacct gaatcaggat attcttctaa tacctggaat
     2461 gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa
     2521 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct
     2581 gtaacatcat tggcaacgct acctttgcca tgtttaagaa acaactctgg cgcatcgggc
     2641 ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta
     2701 tacccatata aatcagcatc catgttggaa tttaatcgcg gcttcgagca agacgtttcc
     2761 cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt
     2821 gttcatgatg atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg
     2881 tgagatcaaa ggatcttc
//
