LOCUS       sequence-436322-        3949 bp    DNA     circular SYN 15-APR-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 3949)
  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 3949)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pTSAR3.1t
FEATURES             Location/Qualifiers
     source          1..3949
                     /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             join(899..901,902..904,905..1618)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(944..965)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1205..1224)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1552..1573
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2069..2097)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2692..2712
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2875..2892)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2875..2891)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2891..2962
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2963..3031,3032..3823)
                     /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(3181..3200)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     3824..3843
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3824..3842
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3931..3949)
                     /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 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
      961 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1021 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1081 cgtgaccacc ctgacctggg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca
     1141 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1201 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1261 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1321 gctggagtac aacgccatca gcgacaacgt ctatatcacc gccgacaagc agaagaacgg
     1381 catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     1441 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1501 cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1561 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaacc
     1621 atggagatat cccaattatt gaaggcctcc ctaacggggg gccttttttt gtttctggtc
     1681 tcccgctagc gaattgccct cgtattaaat gtgtatcgtt tttttacagt aattttaatt
     1741 tgttattcta taataggaat aaatgagctt gactaaaata tgatgtaaaa caaatatact
     1801 tactcggatt acatttcctt gctcactgga atgttgaatt cagtgttatt attgactgca
     1861 gtcacaataa aaaagaattc catagaaaac ctccataaat aaattacaac taacttctgt
     1921 tatgtgtaaa atggaaacta ttaaaactta atatcggaaa tggtaagtga aatttgtata
     1981 aatatacaat tttaaatatt tattctaacg ttctagctag gagaaattaa ccatgaaaag
     2041 atctaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga
     2101 tgttaatggg cacaaatttt ctgtccgtgg agagggtgaa ggtgatgcta caaacggaaa
     2161 actcaccctt aaatttattt gcactactgg aaaactacct gttccgtggc caacacttgt
     2221 cactactttt gcgtatggcc tgcagtgctt tgcacgttat ccggatcaca tgaaacggca
     2281 tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaacgcacta tatctttcaa
     2341 agatgacggg acctacaaga cgcgtgctga agtcaagttt gaaggtgata cccttgttaa
     2401 tcgtatcgag ttaaagggta ttgattttaa agaagatgga aacattcttg gacacaaact
     2461 cgagtacaac tttaactcac acaatgtata catcacggca gacaaacaaa agaatggaat
     2521 caaagctaac ttcaaaattc gccacaacgt tgaagatggt tccgttcaac tagcagacca
     2581 ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct
     2641 gtcgacacaa tctgtccttt cgaaagatcc caacgaaaag cgtgaccaca tggtccttct
     2701 tgagtttgta actgctgctg ggattacaca tggcatggat gaactataca aacggccggc
     2761 ggccaacgat gaaaattatg cgctggccgc ataatctaga actgactcgg taccaaattc
     2821 cagaaaagag gcctcccgaa aggggggcct tttttcgttt tggtcctaag cttaactggc
     2881 cgtcgtttta cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2941 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     3001 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     3061 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     3121 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     3181 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     3241 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3301 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3361 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3421 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3481 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     3541 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3601 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3661 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3721 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3781 acagatcgct gagataggtg cctcactgat taagcattgg taataatacg actcactata
     3841 gggtaactaa ttgaggcgcg cctaccaccg tcaaaaaaaa cggcgctttt tagcgccgtt
     3901 tttatttttc aaccttccta ggtaagtgac cctttagtga gggttaatt
//
