LOCUS       sequence-429627-        3928 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 3928)
  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 3928)
  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: pTSAR3.6t
FEATURES             Location/Qualifiers
     source          1..3928
                     /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"
     CDS             899..1594
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1032..1050)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1300..1319
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2048..2076)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2671..2691
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2854..2871)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2854..2870)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2870..2941
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2942..3010,3011..3802)
                     /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(3160..3179)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     3803..3822
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3803..3821
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3910..3928)
                     /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 cggtacccgg ggatcctcta gagtcgacct gcaggtagga ggtaaataat
      901 ggtgagcaag ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc acatggaggg
      961 ctccatgaac ggtcacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     1021 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttct cctgggacat
     1081 cctgtcccct cagttcatgt acggctccag ggccttcacc aagcaccccg ccgacatccc
     1141 cgactactat aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     1201 ggacggcggc gccgtgaccg tgacccagga cacctccctg gaggacggca ccctgatcta
     1261 caaggtgaag ctccgcggca ccaacttccc tcctgacggc cccgtaatgc agaagaagac
     1321 aatgggctgg gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc tgaagggcga
     1381 cattaagatg gccctgcgcc tgaaggacgg cggccgctac ctggcggact tcaagaccac
     1441 ctacaaggcc aagaagcccg tgcagatgcc cggcgcctac aacgtcgacc gcaagttgga
     1501 catcacctcc cacaacgagg actacaccgt ggtggaacag tacgaacgct ccgagggccg
     1561 ccactccacc ggcggcatgg acgagctgta caagtaacca tggagatatc ccaattattg
     1621 aaggcctccc taacgggggg cctttttttg tttctggtct cccgctagcg aattgccctc
     1681 gtattaaatg tgtatcgttt ttttacagta attttaattt gttattctat aataggaata
     1741 aatgagcttg actaaaatat gatgtaaaac aaatatactt actcggatta catttccttg
     1801 ctcactggaa tgttgaattc agtgttatta ttgactgcag tcacaataaa aaagaattcc
     1861 atagaaaacc tccataaata aattacaact aacttctgtt atgtgtaaaa tggaaactat
     1921 taaaacttaa tatcggaaat ggtaagtgaa atttgtataa atatacaatt ttaaatattt
     1981 attctaacgt tctagctagg agaaattaac catgaaaaga tctaaaggag aagaactttt
     2041 cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc
     2101 tgtccgtgga gagggtgaag gtgatgctac aaacggaaaa ctcaccctta aatttatttg
     2161 cactactgga aaactacctg ttccgtggcc aacacttgtc actacttttg cgtatggcct
     2221 gcagtgcttt gcacgttatc cggatcacat gaaacggcat gactttttca agagtgccat
     2281 gcccgaaggt tatgtacagg aacgcactat atctttcaaa gatgacggga cctacaagac
     2341 gcgtgctgaa gtcaagtttg aaggtgatac ccttgttaat cgtatcgagt taaagggtat
     2401 tgattttaaa gaagatggaa acattcttgg acacaaactc gagtacaact ttaactcaca
     2461 caatgtatac atcacggcag acaaacaaaa gaatggaatc aaagctaact tcaaaattcg
     2521 ccacaacgtt gaagatggtt ccgttcaact agcagaccat tatcaacaaa atactccaat
     2581 tggcgatggc cctgtccttt taccagacaa ccattacctg tcgacacaat ctgtcctttc
     2641 gaaagatccc aacgaaaagc gtgaccacat ggtccttctt gagtttgtaa ctgctgctgg
     2701 gattacacat ggcatggatg aactatacaa acggccggcg gccaacgatg aaaattatgc
     2761 gctggccgca taatctagaa ctgactcggt accaaattcc agaaaagagg cctcccgaaa
     2821 ggggggcctt ttttcgtttt ggtcctaagc ttaactggcc gtcgttttac attcaaatat
     2881 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2941 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     3001 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     3061 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     3121 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     3181 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     3241 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     3301 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     3361 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     3421 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     3481 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     3541 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     3601 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     3661 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     3721 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3781 ctcactgatt aagcattggt aataatacga ctcactatag ggtaactaat tgaggcgcgc
     3841 ctaccaccgt caaaaaaaac ggcgcttttt agcgccgttt ttatttttca accttcctag
     3901 gtaagtgacc ctttagtgag ggttaatt
//
