LOCUS       sequence-424903-        3881 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for Type III secretion system activity in 
            Shigella flexneri.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3881)
  AUTHORS   Manigat FO, Connell LB, Stewart BN, LePabic AR, Tessier CJG, Emlaw 
            JR, Calvert ND, Rossl A, Shuhendler AJ, daCosta CJB, Campbell-Valois
            FX
  TITLE     pUdOs: Concise Plasmids for Bacterial and Mammalian Cells.
  JOURNAL   ACS Synth Biol. 2024 Jan 18. doi: 10.1021/acssynbio.3c00408.
  PUBMED    38235654
REFERENCE   2  (bases 1 to 3881)
  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: "ACS Synth 
            Biol. 2024 Jan 18. doi: 10.1021/acssynbio.3c00408."
COMMENT     Sequence Label: pTSAR3.7t
FEATURES             Location/Qualifiers
     source          1..3881
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      23..611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     512..531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     630..646
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     630..646
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     RBS             861..869
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             873..1550
                     /codon_start=1
                     /product="far-red noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2009)"
                     /label=E2-Crimson
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGVGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFFYGSKAYIKHPADIPDYLKQSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTLIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERNYPRDGVLKGENHM
                     ALKLKGGGHYLCEFKSIYMAKKPVKLPGYHYVDYKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     1271..1290
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2001..2029)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2624..2644
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2807..2824)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2807..2823)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2823..2894
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2895..2963,2964..3755)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3113..3132)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     3756..3775
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3756..3774
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3863..3881)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
       61 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
      121 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
      181 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
      241 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
      301 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
      361 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
      421 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
      481 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
      541 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      601 gcctatggaa aaacgccagc aacgcggccc aggaaacagc tatgaccatg cccgggtaag
      661 tgactagcgg aaacacagaa aaaagcccgc acctgacagt gcgggctttt tttttcgacc
      721 aaaggctccg gcggccgcga ttttttcgca tatttttctt gcaaagttgg gttgagctgg
      781 ctagattagc cagccaatct tttgtatgtc tgtgcgttcg agtaactagc taagattacg
      841 aattcgagct cggtaccggg taaggaggta aaatggatag cactgagaac gtcatcaagc
      901 ccttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
      961 agggcgtggg cgagggcaag ccctacgagg gcacccagac cgccaagctg caagtgacca
     1021 agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttcttc tacggctcca
     1081 aggcgtacat caagcacccc gccgacatcc ccgactacct caagcagtcc ttccccgagg
     1141 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg
     1201 actcctccct gcaggacggc accctcatct accacgtgaa gttcatcggc gtgaacttcc
     1261 cctccgacgg ccccgtaatg cagaagaaga ctctgggctg ggagccctcc actgagcgca
     1321 actacccccg cgacggcgtg ctgaagggcg agaaccacat ggcgctgaag ctgaagggcg
     1381 gcggccacta cctgtgtgag ttcaagtcca tctacatggc caagaagccc gtgaagctgc
     1441 ccggctacca ctacgtggac tacaagctcg acatcacctc ccacaacgag gactacaccg
     1501 tggtggagca gtacgagcgc gccgaggccc gccaccacct gttccagtag ccatggagat
     1561 atcccaatta ttgaaggcct ccctaacggg gggccttttt ttgtttctgg tctcccgcta
     1621 gcgaattgcc ctcgtattaa atgtgtatcg tttttttaca gtaattttaa tttgttattc
     1681 tataatagga ataaatgagc ttgactaaaa tatgatgtaa aacaaatata cttactcgga
     1741 ttacatttcc ttgctcactg gaatgttgaa ttcagtgtta ttattgactg cagtcacaat
     1801 aaaaaagaat tccatagaaa acctccataa ataaattaca actaacttct gttatgtgta
     1861 aaatggaaac tattaaaact taatatcgga aatggtaagt gaaatttgta taaatataca
     1921 attttaaata tttattctaa cgttctagct aggagaaatt aaccatgaaa agatctaaag
     1981 gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt gatgttaatg
     2041 ggcacaaatt ttctgtccgt ggagagggtg aaggtgatgc tacaaacgga aaactcaccc
     2101 ttaaatttat ttgcactact ggaaaactac ctgttccgtg gccaacactt gtcactactt
     2161 ttgcgtatgg cctgcagtgc tttgcacgtt atccggatca catgaaacgg catgactttt
     2221 tcaagagtgc catgcccgaa ggttatgtac aggaacgcac tatatctttc aaagatgacg
     2281 ggacctacaa gacgcgtgct gaagtcaagt ttgaaggtga tacccttgtt aatcgtatcg
     2341 agttaaaggg tattgatttt aaagaagatg gaaacattct tggacacaaa ctcgagtaca
     2401 actttaactc acacaatgta tacatcacgg cagacaaaca aaagaatgga atcaaagcta
     2461 acttcaaaat tcgccacaac gttgaagatg gttccgttca actagcagac cattatcaac
     2521 aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac ctgtcgacac
     2581 aatctgtcct ttcgaaagat cccaacgaaa agcgtgacca catggtcctt cttgagtttg
     2641 taactgctgc tgggattaca catggcatgg atgaactata caaacggccg gcggccaacg
     2701 atgaaaatta tgcgctggcc gcataatcta gaactgactc ggtaccaaat tccagaaaag
     2761 aggcctcccg aaaggggggc cttttttcgt tttggtccta agcttaactg gccgtcgttt
     2821 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2881 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2941 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3001 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3061 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3121 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3181 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3241 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3301 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3361 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3421 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3481 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3541 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3601 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3661 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3721 ctgagatagg tgcctcactg attaagcatt ggtaataata cgactcacta tagggtaact
     3781 aattgaggcg cgcctaccac cgtcaaaaaa aacggcgctt tttagcgccg tttttatttt
     3841 tcaaccttcc taggtaagtg accctttagt gagggttaat t
//
