LOCUS       sequence-422904-        3914 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 3914)
  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 3914)
  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: pTSAR3.4t
FEATURES             Location/Qualifiers
     source          1..3914
                     /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..1583
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1018..1036)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1286..1305
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2034..2062)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2657..2677
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2840..2857)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2840..2856)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2856..2927
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2928..2996,2997..3788)
                     /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(3146..3165)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     3789..3808
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3789..3807
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3896..3914)
                     /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 aaatggtgag caagggcgag gaggataaca
      901 tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc
      961 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
     1021 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt
     1081 tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc
     1141 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
     1201 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
     1261 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg
     1321 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc
     1381 tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga
     1441 agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca
     1501 acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg
     1561 gcatggacga gctgtacaag tagccatgga gatatcccaa ttattgaagg cctccctaac
     1621 ggggggcctt tttttgtttc tggtctcccg ctagcgaatt gccctcgtat taaatgtgta
     1681 tcgttttttt acagtaattt taatttgtta ttctataata ggaataaatg agcttgacta
     1741 aaatatgatg taaaacaaat atacttactc ggattacatt tccttgctca ctggaatgtt
     1801 gaattcagtg ttattattga ctgcagtcac aataaaaaag aattccatag aaaacctcca
     1861 taaataaatt acaactaact tctgttatgt gtaaaatgga aactattaaa acttaatatc
     1921 ggaaatggta agtgaaattt gtataaatat acaattttaa atatttattc taacgttcta
     1981 gctaggagaa attaaccatg aaaagatcta aaggagaaga acttttcact ggagttgtcc
     2041 caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc cgtggagagg
     2101 gtgaaggtga tgctacaaac ggaaaactca cccttaaatt tatttgcact actggaaaac
     2161 tacctgttcc gtggccaaca cttgtcacta cttttgcgta tggcctgcag tgctttgcac
     2221 gttatccgga tcacatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg
     2281 tacaggaacg cactatatct ttcaaagatg acgggaccta caagacgcgt gctgaagtca
     2341 agtttgaagg tgataccctt gttaatcgta tcgagttaaa gggtattgat tttaaagaag
     2401 atggaaacat tcttggacac aaactcgagt acaactttaa ctcacacaat gtatacatca
     2461 cggcagacaa acaaaagaat ggaatcaaag ctaacttcaa aattcgccac aacgttgaag
     2521 atggttccgt tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg
     2581 tccttttacc agacaaccat tacctgtcga cacaatctgt cctttcgaaa gatcccaacg
     2641 aaaagcgtga ccacatggtc cttcttgagt ttgtaactgc tgctgggatt acacatggca
     2701 tggatgaact atacaaacgg ccggcggcca acgatgaaaa ttatgcgctg gccgcataat
     2761 ctagaactga ctcggtacca aattccagaa aagaggcctc ccgaaagggg ggcctttttt
     2821 cgttttggtc ctaagcttaa ctggccgtcg ttttacattc aaatatgtat ccgctcatga
     2881 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     2941 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3001 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3061 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3121 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3181 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3241 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3301 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3361 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3421 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3481 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3541 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3601 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3661 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3721 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3781 attggtaata atacgactca ctatagggta actaattgag gcgcgcctac caccgtcaaa
     3841 aaaaacggcg ctttttagcg ccgtttttat ttttcaacct tcctaggtaa gtgacccttt
     3901 agtgagggtt aatt
//
