LOCUS       sequence-405410-        7645 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for SPI2 induction in Salmonella.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7645)
  AUTHORS   Stevenin V, Coipan CE, Duijster JW, van Elsland DM, Voogd L, Bigey 
            L, van Hoek AHAM, Wijnands LM, Janssen L, Akkermans JJLL, 
            Neefjes-Borst A, Franz E, Mughini-Gras L, Neefjes J
  TITLE     Multi-omics analyses of cancer-linked clinical salmonellae reveal 
            bacterial-induced host metabolic shift and mTOR-dependent cell 
            transformation.
  JOURNAL   Cell Rep. 2024 Nov 1;43(11):114931. doi: 
            10.1016/j.celrep.2024.114931.
  PUBMED    39488829
REFERENCE   2  (bases 1 to 7645)
  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: "Cell Rep.";
            date: "2024-11-1"; volume: "43(11):114931. doi"; pages: " 
            10.1016/j.celrep.2024.114931"
COMMENT     Sequence Label: pFPV25-cR-2iG
FEATURES             Location/Qualifiers
     source          1..7645
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(771..789)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(821..841)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1039..1058
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1233..1256
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     oriT            1926..2013
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     primer_bind     complement(3326..3343)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3497..4085)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3577..3596)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4395..5189)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4395..5189)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4506..4525)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5116..5135
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5190..5294)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5362..5380
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(5404..5424)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             complement(5425..6138)
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(5465..5485)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     6080..6108
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 gtacccgggg atcccggagt tcatgcgtga tgcaatgaaa gtgccgttct acttcggtgg
       61 gacctcactg cttatcgttg ttgtcgtgat tatggacttt atggctcaag tgcaaactct
      121 gatgatgtcc agtcagtatg agtctgcatt gaagaaggcg aacctgaaag gctacggccg
      181 ttaattggtc gcctgagaag ttacggagag taaaaatgaa agttcgtgct tccgtcaaga
      241 aattatgccg taactgcaaa atcgttaagc gtgatggtgt catccgtgtg atttgcagtg
      301 ccgagccgaa gcataaacag cgccaaggct gattttttcg catatttttc ttgcaaagtt
      361 gggttgagct ggctagatta gccagccaat cttttgtatg tctgtacgtt tccatttgag
      421 tatcctgaaa acgggctttt cagcatggta cgtacatatt aaatagtagg agtgcatagt
      481 ggcccgtata gcaggcatta acattcctga tcagaaacac gccgtgatcg cgttaacttc
      541 gatctacggt gtcggcaaga cccgttctaa agccatcctg gctgcagcgg gtatcgctga
      601 aaatgttaag atcctctaga tttaagaagg agatatacat atggcctcca ccgaggacgt
      661 catcaaggag ttcatgcgct tcaaggtgcg catggagggc tccgtgaacg gccacgagtt
      721 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
      781 ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc agttccagta
      841 cggctccaag gtgtacgtga agcaccccgc cgacatcccc gactacaaga agctgtcctt
      901 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
      961 gacccaggac tcctccctgc aggacggctg cttcatctac aaggtgaagt tcatcggcgt
     1021 gaacttcccc tccgacggcc ccgtaatgca gaagaagact atgggctggg agccctccac
     1081 cgagcgcctg tacccccgcg acggcgtgct gaagggcgag atccacaagg ccctgaagct
     1141 gaaggacggc ggccactacc tggtggagtt caagtccatc tacatggcca agaagcccgt
     1201 gcagctgccc ggctactact acgtggactc caagctggac atcacctccc acaacgagga
     1261 ctacaccatc gtggagcagt acgagcgcac cgagggccgc taccacctgt tcctgtagaa
     1321 gctttaatgc ggtagtttat cacagttaaa ttgctaacgc agtcaggcac cgtgtatgaa
     1381 atctaacaat gcgctcatcg tcatcctcgg caccgtcacc ctggatgctg taggcatagg
     1441 cttggttatg ccggtactgc cgggcctctt gcgggatatc gtccattccg acagcatcgc
     1501 cagtcactat ggcgtgctgc tagcgctttt agccgcttta gcggcctttc cccctacccg
     1561 aagggtgggg gcgcgtgtgc agccccgcag ggcctgtctc ggtcgatcat tcagcccggc
     1621 tcatccttct ggcgtggcgg cagaccgaac aaggcgcggt cgtggtcgcg ttcaaggtac
     1681 gcatccattg ccgccatgag ccgatcctcc ggccactcgc tgctgttcac cttggccaaa
     1741 atcatggccc ccaccagcac cttgcgcctt gtttcgttct tgcgctcttg ctgctgttcc
     1801 cttgcccgca cccgctgaat ttcggcattg attcgcgctc gttgttcttc gagcttggcc
     1861 agccgatccg ccgccttgtt gctcccctta accatcttga caccccattg ttaatgtgct
     1921 gtctcgtagg ctatcatgga ggcacagcgg cggcaatccc gaccctactt tgtaggggag
     1981 ggggcactta ccggtttctc ttcgagaaac tggcctaacg gccacccttc gggcggtgcg
     2041 ctctccgagg gccattgcat ggagccgaaa agcaaaagca acagcgaggc agcatggcga
     2101 tttatcacct tacggcgaaa accggcagca ggtcgggcgg ccaatcggcc agggccaagg
     2161 ccgactacat ccagcgcgaa ggcaagtatg cccgcgacat ggatgaagtc ttgcacgccg
     2221 aatccgggca catgccggag ttcgtcgagc ggcccgccga ctactgggat gctgccgacc
     2281 tgtatgaacg cgccaatggg cggctgttca aggaggtcga atttgccctg ccggtcgagc
     2341 tgaccctcga ccagcagaag gcgctggcgt ccgagttcgc ccagcacctg accggtgccg
     2401 agcgcctgcc gtatacgctg gccatccatg ccggtggcgg cgagaacccg cactgccacc
     2461 tgatgatctc cgagcggatc aatgacggca tcgagcggcc cgccgctcag tggttcaagc
     2521 ggtacaacgg caagaccccg gagaagggcg gggcacagaa gaccgaagcg ctcaagccca
     2581 aggcatggct tgagcagacc cgcgaggcat gggccgacca tgccaaccgg gcattagagc
     2641 gggctggcca cgacgcccgc attgaccaca gaacacttga ggcgcagggc atcgagcgcc
     2701 tgcccggtgt tcacctgggg ccgaacgtgg tggagatgga aggccggggc atccgcaccg
     2761 accgggcaga cgtggccctg aacatcgaca ccgccaacgc ccagatcatc gacttacagg
     2821 aataccggga ggcaatagac catgaacgca atcgacagag tgaagaaatc cagaggcatc
     2881 aacgagttag cggagcagat cgaaccgctg gcccagagca tggcgacact ggccgacgaa
     2941 gcccggcagg tcatgagcca gacccagcag gccagcgagg cgcaggcggc ggagtggctg
     3001 aaagcccagc gccagacagg ggcggcatgg gtggagctgg ccaaagagtt gcgggaggta
     3061 gccgccgagg tgagcagcgc cgcgcagagc gcccggagcg cgtcgcgggg gtggcactgg
     3121 aagctatggc taaccgtgat gctggcttcc atgatgccta cggtggtgct gctgatcgca
     3181 tcgttgctct tgctcgacct gacgccactg acaaccgagg acggctcgat ctggctgcgc
     3241 ttggtggccc gatgaagaac gacaggactt tgcaggccat aggccgacag ctcaaggcca
     3301 tgggctgtga gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3361 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3421 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3481 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3541 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3601 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3661 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3721 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3781 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3841 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3901 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     3961 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4021 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4081 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4141 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4201 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4261 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4321 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4381 ctgcaatgat taattcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa
     4441 tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct
     4501 tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg
     4561 ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca
     4621 tcgccatggg tcacgacgag atcctcgccg tcgggcatgc tcgccttgag cctggcgaac
     4681 agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg
     4741 gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag
     4801 gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg
     4861 gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag
     4921 tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc
     4981 agccacgata gccgcgctgc ctcgtcttgc agttcattca gggcaccgga caggtcggtc
     5041 ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag
     5101 ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa
     5161 cctgcgtgca atccatcttg ttcaatcata ctcttccttt ttcaatatta ttgaagcatt
     5221 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5281 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     5341 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct tcaagaattc
     5401 tcagtggtgg tggtggtggt ggtgtttgta tagttcatcc atgccatgtg taatcccagc
     5461 agctgttaca aactcaagaa ggaccatgtg gtctctcttt tcgttgggat ctttcgaaag
     5521 ggcagattgt gtggacaggt aatggttgtc tggtaaaagg acagggccat cgccaattgg
     5581 agtattttgt tgataatggt ctgctagttg aacgcttcca tcttcaatgt tgtgtctaat
     5641 tttgaagtta actttgattc cattcttttg tttgtctgcc atgatgtata cattgtgtga
     5701 gttatagttg tattccaact tgtgtccaag aatgtttcca tcttctttaa aatcaatacc
     5761 ttttaactcg attctattaa caagtgtatc accttcaaac ttgacttcag cacgtgtctt
     5821 atagttcccg tcatctttga aaaatatagt tctttcctgt acataacctt cgggcatggc
     5881 actcttgaaa aagtcatgcc gtttcatatg atctgggtat cttgaaaagc attgaacacc
     5941 ataagtcaaa gtagtgacaa gtgttggcca tggaacaggt agttttccag tagtgcaaat
     6001 aaatttaagg gtaagttttc cgtatgtcgc atcaccttca ccctctccac tgacagaaaa
     6061 tttgtgccca ttaacatcac catctaattc aacaagaatt gggacaactc cagtgaaaag
     6121 ttcttctcct ttactcatac agccgttcta gatgcccttt cagcaagctg ttgactcacc
     6181 ttagcccgga tttggtgaaa ttttttaaga cagtgattta gctgcatgta actttgctct
     6241 aaatcacgat attcactaaa cgcagccttt ttctttatca tcattcccct ccatatacac
     6301 gatagataat taacgtgcta actaagagcc tatcccatta gggctatttt acttgccatt
     6361 ttgaacctgg gcagtgctcg ccaaaacgcg ttagcgtttt gaacgccgct agcggcggcc
     6421 cgaagggcga gcgtagcgag tcaaacctca cgtactacgt gtacgctgcg gtttttgcgc
     6481 gctatccgtg tccaaactgg ctgcgccaat taacgcctgg tgggataggc tctaagatat
     6541 ttttacttta ctcttgctca ctcactacaa gtgcgctgtt atggtaacga taataaataa
     6601 tgttgatgat attttcggcc tgctcgatcg cttcaagcaa taaggtgttt tgctgatatt
     6661 gctgcggatc ctgtaacttg ccgttgttct cttctaactg tttccgggca gcccttaatt
     6721 gtaaaattat gcctttggcc tcttcacgcg aatgaagcag caaatcttct aaccgggtca
     6781 aagttgtcat tttccactca cttaaaatct aatggatagt taatcaaagt atcataatgt
     6841 ttaatcgtta ccacatcggc actcagatgg acaatttctc ccccattggg taacaatgcc
     6901 cctacacgta aacgctcttt attcgtcagt aataagtaat taccatggcg actctgtaca
     6961 aagccagcca ctggcgcagg cagatacttg ctttcatcat gggaaggcgc aatatcctga
     7021 taaattaaag aaagagcggg atcctttttc tttaatgctg ctaacgtttc ttgcaaaatg
     7081 cgttgatgag attcatccag tacaccactg ataacaaaag agcgccgcat tggcgtaaca
     7141 ttgacaagcc ccactaaacc gttctctatt atcgcagaaa taatatcatc cccctgagac
     7201 tgatgagagt gactaatctg ccagtgcaat aacccgggaa tatctgcaag taatggttga
     7261 accttgcgcc attgctgatc catttgtata tcatcatgaa ttaacacgct ccccggccct
     7321 tcgctggata cttcagcatg cgggtaaccc atttttatca aaacatcctg cacttctcgt
     7381 accaataagt catcacagat tacaccatcc cgatacatga ccccccatga ttcgagagtc
     7441 gctctcacct tttgcatctg ttcgcttgac gagcaataac cggacaactg caggctgcca
     7501 tcttctttcc attgcgcccg cacataatga atattgcttt tgtctaataa aaacttaacc
     7561 cgcaaaggta agtcatttac cgtttcaggc tgaccactaa tacttaacag gacacccatt
     7621 ccaccgatga aaatcaagaa tacgg
//
