LOCUS       sequence-424838-        3916 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 3916)
  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 3916)
  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: pTSAR3.3t
FEATURES             Location/Qualifiers
     source          1..3916
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(61..77)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(252..268)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(287..875)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(367..386)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        898..916
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1004..1023)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1005..1023)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(join(1024..1815,1816..1884))
                     /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     1647..1666
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1885..1956)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1955..1972
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1956..1972
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2135..2155)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     2750..2778
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     CDS             complement(3229..3906)
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASTEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEPSTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(3291..3314)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(3489..3508)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3706..3726
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     3758..3776
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
ORIGIN
        1 ccttcttaaa ggatccccgg gtaccgagct cgaattcgta atcttagcta gttactcgag
       61 gtcatagctg tttcctgtgt acgcacagac atacaaaaga ttggctggct aatctagcca
      121 gctcaaccca actttgcaag aaaaatatgc gaaaaaatcg cggccgccgg agcctttggt
      181 cgaaaaaaaa agcccgcact gtcaggtgcg ggcttttttc tgtgtttccg ctagtcactt
      241 acccgggcat ggtcatagct gtttcctggg ccgcgttgct ggcgtttttc cataggctcc
      301 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
      361 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
      421 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
      481 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
      541 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
      601 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
      661 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
      721 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
      781 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
      841 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctaaat
      901 taaccctcac taaagggtca cttacctagg aaggttgaaa aataaaaacg gcgctaaaaa
      961 gcgccgtttt ttttgacggt ggtaggcgcg cctcaattag ttaccctata gtgagtcgta
     1021 ttattaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     1081 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     1141 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     1201 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     1261 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     1321 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     1381 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     1441 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     1501 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     1561 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     1621 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     1681 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     1741 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     1801 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     1861 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     1921 gggttattgt ctcatgagcg gatacatatt tgaatgtaaa acgacggcca gttaagctta
     1981 ggaccaaaac gaaaaaaggc ccccctttcg ggaggcctct tttctggaat ttggtaccga
     2041 gtcagttcta gattatgcgg ccagcgcata attttcatcg ttggccgccg gccgtttgta
     2101 tagttcatcc atgccatgtg taatcccagc agcagttaca aactcaagaa ggaccatgtg
     2161 gtcacgcttt tcgttgggat ctttcgaaag gacagattgt gtcgacaggt aatggttgtc
     2221 tggtaaaagg acagggccat cgccaattgg agtattttgt tgataatggt ctgctagttg
     2281 aacggaacca tcttcaacgt tgtggcgaat tttgaagtta gctttgattc cattcttttg
     2341 tttgtctgcc gtgatgtata cattgtgtga gttaaagttg tactcgagtt tgtgtccaag
     2401 aatgtttcca tcttctttaa aatcaatacc ctttaactcg atacgattaa caagggtatc
     2461 accttcaaac ttgacttcag cacgcgtctt gtaggtcccg tcatctttga aagatatagt
     2521 gcgttcctgt acataacctt cgggcatggc actcttgaaa aagtcatgcc gtttcatgtg
     2581 atccggataa cgtgcaaagc actgcaggcc atacgcaaaa gtagtgacaa gtgttggcca
     2641 cggaacaggt agttttccag tagtgcaaat aaatttaagg gtgagttttc cgtttgtagc
     2701 atcaccttca ccctctccac ggacagaaaa tttgtgccca ttaacatcac catctaattc
     2761 aacaagaatt gggacaactc cagtgaaaag ttcttctcct ttagatcttt tcatggttaa
     2821 tttctcctag ctagaacgtt agaataaata tttaaaattg tatatttata caaatttcac
     2881 ttaccatttc cgatattaag ttttaatagt ttccatttta cacataacag aagttagttg
     2941 taatttattt atggaggttt tctatggaat tcttttttat tgtgactgca gtcaataata
     3001 acactgaatt caacattcca gtgagcaagg aaatgtaatc cgagtaagta tatttgtttt
     3061 acatcatatt ttagtcaagc tcatttattc ctattataga ataacaaatt aaaattactg
     3121 taaaaaaacg atacacattt aatacgaggg caattcgcta gcgggagacc agaaacaaaa
     3181 aaaggccccc cgttagggag gccttcaata attgggatat ctccatggct acaggaacag
     3241 gtggtggcgg ccctcggtgc gttcgtactg ctccacgatg gtgtagtcct cgttgtggga
     3301 ggtgatgtcc agcttggagt ccacgtagta gtagccgggc agttgcacgg gcttcttggc
     3361 catgtagatg gacttgaact ccaccaggta gtggccgccg tccttcagct tcagggcctt
     3421 gtggatctcg cccttcagca cgccgtcgcg ggggtacagg cgctcggtgg agggctccca
     3481 gcccatagtc ttcttctgca ttacggggcc gtcggagggg aagttcacgc cgatgaactt
     3541 caccttgtag atgaagcagc cgtcctgcag ggaggagtcc tgggtcacgg tcaccacgcc
     3601 gccgtcctcg aagttcatca cgcgctccca cttgaagccc tcggggaagg acagcttctt
     3661 gtagtcgggg atgtcggcgg ggtgcttcac gtacaccttg gagccgtact ggaactgggg
     3721 ggacaggatg tcccaggcga agggcagggg gccgcccttg gtcaccttca gcttggcggt
     3781 ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg atctcgaact cgtggccgtt
     3841 cacggagccc tccatgcgca ctttgaagcg catgaactcc ttgatgacgt cctcggtgga
     3901 tgccatatgt atatct
//
