LOCUS       sequence-422906-        3923 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 3923)
  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 3923)
  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: pTSAR3.5t
FEATURES             Location/Qualifiers
     source          1..3923
                     /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             join(873..875,876..878,879..1592)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(918..939)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2043..2071)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2666..2686
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2849..2866)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2849..2865)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2865..2936
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2937..3005,3006..3797)
                     /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(3155..3174)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     3798..3817
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3798..3816
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3905..3923)
                     /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 gagctgttca
      901 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
      961 tgaggggcga gggcgagggc gatgccacca acggcaagct gaccctgaag ttcatctgca
     1021 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgagc cacggcgtgc
     1081 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1141 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcacc tacaagaccc
     1201 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcgtcg
     1261 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaacttc aacagccaca
     1321 acatctatat catggccgtc aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     1381 acaacgtgga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1441 gcgacggccc cgtgctgctg cccgacagcc actacctgag cacccagtcc gtgctgagca
     1501 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttccgcacc gccgccggga
     1561 tcactctcgg catggacgag ctgtacaagt aaccatggag atatcccaat tattgaaggc
     1621 ctccctaacg gggggccttt ttttgtttct ggtctcccgc tagcgaattg ccctcgtatt
     1681 aaatgtgtat cgttttttta cagtaatttt aatttgttat tctataatag gaataaatga
     1741 gcttgactaa aatatgatgt aaaacaaata tacttactcg gattacattt ccttgctcac
     1801 tggaatgttg aattcagtgt tattattgac tgcagtcaca ataaaaaaga attccataga
     1861 aaacctccat aaataaatta caactaactt ctgttatgtg taaaatggaa actattaaaa
     1921 cttaatatcg gaaatggtaa gtgaaatttg tataaatata caattttaaa tatttattct
     1981 aacgttctag ctaggagaaa ttaaccatga aaagatctaa aggagaagaa cttttcactg
     2041 gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtcc
     2101 gtggagaggg tgaaggtgat gctacaaacg gaaaactcac ccttaaattt atttgcacta
     2161 ctggaaaact acctgttccg tggccaacac ttgtcactac ttttgcgtat ggcctgcagt
     2221 gctttgcacg ttatccggat cacatgaaac ggcatgactt tttcaagagt gccatgcccg
     2281 aaggttatgt acaggaacgc actatatctt tcaaagatga cgggacctac aagacgcgtg
     2341 ctgaagtcaa gtttgaaggt gatacccttg ttaatcgtat cgagttaaag ggtattgatt
     2401 ttaaagaaga tggaaacatt cttggacaca aactcgagta caactttaac tcacacaatg
     2461 tatacatcac ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa attcgccaca
     2521 acgttgaaga tggttccgtt caactagcag accattatca acaaaatact ccaattggcg
     2581 atggccctgt ccttttacca gacaaccatt acctgtcgac acaatctgtc ctttcgaaag
     2641 atcccaacga aaagcgtgac cacatggtcc ttcttgagtt tgtaactgct gctgggatta
     2701 cacatggcat ggatgaacta tacaaacggc cggcggccaa cgatgaaaat tatgcgctgg
     2761 ccgcataatc tagaactgac tcggtaccaa attccagaaa agaggcctcc cgaaaggggg
     2821 gccttttttc gttttggtcc taagcttaac tggccgtcgt tttacattca aatatgtatc
     2881 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     2941 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3001 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3061 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3121 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3181 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3241 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3301 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3361 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3421 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3481 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3541 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3601 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3661 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3721 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3781 tgattaagca ttggtaataa tacgactcac tatagggtaa ctaattgagg cgcgcctacc
     3841 accgtcaaaa aaaacggcgc tttttagcgc cgtttttatt tttcaacctt cctaggtaag
     3901 tgacccttta gtgagggtta att
//
