LOCUS       sequence-389647-        6720 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacteria reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6720)
  AUTHORS   Schulte M, Olschewski K, Hensel M
  TITLE     The protected physiological state of intracellular Salmonella 
            enterica persisters reduces host cell-imposed stress.
  JOURNAL   Commun Biol. 2021 May 4;4(1):520. doi: 10.1038/s42003-021-02049-6.
  PUBMED    33947954
REFERENCE   2  (bases 1 to 6720)
  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: "Commun 
            Biol."; date: "2021-05-4"; volume: "4(1):520. doi"; pages: " 
            10.1038/s42003-021-02049-6"
COMMENT     Sequence Label: p5300
FEATURES             Location/Qualifiers
     source          1..6720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..56
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    7..25
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    37..55
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             68..79
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             83..760
                     /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(213..231)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(263..283)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     481..500
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     675..698
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             2132..2323
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2317..2339)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2425..2565
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2480..2499
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2580..2597)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2751..3339)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2831..2850)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3510..4301,4302..4370))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4133..4152
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4371..4462)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(4481..4508)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4600..4686)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4839..4856
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(4898..5611)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(5950..5969)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(6079..6705)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVISSALELLNEVGIEGLTTRKLAQRLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGKSWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     6610..6629
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
ORIGIN
        1 gttgacactc tatcattgat agagttattt taccactccc tatcagtgat agagaaaagt
       61 gaacgggaaa gaggagaaaa gtatggcatc caccgaggac gtcatcaagg agttcatgcg
      121 cttcaaagtg cgcatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
      181 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc
      241 cctgcccttc gcctgggaca tcctgtcccc ccagttccag tacggctcca aggtgtacgt
      301 gaagcacccc gccgacatcc ccgactacaa gaagctgtcc ttccccgagg gcttcaagtg
      361 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
      421 gcaggacggc tgcttcatct acaaggtgaa gttcatcggc gtgaacttcc cctccgacgg
      481 ccccgtaatg cagaagaaga ctatgggctg ggagccctcc accgagcgcc tgtacccccg
      541 cgacggcgtg ctgaagggcg agatccacaa ggccctgaag ctgaaggacg gcggccacta
      601 cctggtggag ttcaagtcca tctacatggc caagaagccc gtgcaactgc ccggctacta
      661 ctacgtggac tccaagctgg acatcacctc ccacaacgag gactacacca tcgtggagca
      721 gtacgaacgc accgagggcc gccaccacct gttcctgtag ccatggatcg atagctggtc
      781 gacaaccatt ccttgcggcg gcggtgctca acggcctcaa cctactactg ggctgcttcc
      841 taatgcagga gtcgcataag ggagagcgtc gaccgatgcc cttgagagcc ttcaacccag
      901 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
      961 ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc
     1021 gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg
     1081 ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca
     1141 ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc gcgacgcgag
     1201 gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg atgcccgcgt
     1261 tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt caaggatcgc
     1321 tcgcggctct taccagccta acttcgatca ttggaccgct gatcgtcacg gcgatttatg
     1381 ccgcctcggc gagcacatgg aacgggttgg catggattgt aggcgccgcc ctataccttg
     1441 tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc cacctcgacc tgaatggaag
     1501 ccggcggcac ctcgctaacg gattcaccac tccaagaatt ggagccaatc aattcttgcg
     1561 gagaactgtg aatgcgcaaa ccaacccttg gcagaacata tccatcgcgt ccgccatctc
     1621 cagcagccgc acgcggcgca tctcgggcag cgttgggtcc tggccacggg tgcgcatgat
     1681 cgtgctcctg tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa
     1741 tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg
     1801 agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc
     1861 agcgccctgc accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg
     1921 aacacctaca tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc
     1981 ccgccgcatc cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc
     2041 atcatcagta acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat
     2101 gaacagaaat cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa
     2161 catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga
     2221 cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg
     2281 cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     2341 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     2401 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     2461 gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     2521 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc
     2581 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2641 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2701 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2761 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2821 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2881 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2941 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3001 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3061 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3121 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3181 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3241 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3301 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3361 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3421 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3481 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3541 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3601 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3661 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3721 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3781 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3841 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3901 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3961 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4021 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4081 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc
     4141 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4201 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4261 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4321 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4381 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4441 tgtatttaga aaaataaaca aaagagtttg tagaaacgca aaaaggccat ccgtcaggat
     4501 ggccttctgc ttaatttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc
     4561 gggccgttgc ttcgcaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc
     4621 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct ttcgactgag cctttcgttt
     4681 tatttgatgc ctggcagttc cctactctcg catggggaga ccccacacta ccatcggcgc
     4741 tacggcgttt cacttctgag ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc
     4801 caggcaaatt ctgttttatc agaccgcttc tgcgttctga tttaatctgt atcaggctga
     4861 aaatcttctc tcatccgcca aaacagaagt tttattattt atacagttca tccatgccat
     4921 gcgtaatacc cgccgcggtc acaaattcca gcagcaccat atggtcgcgt ttttcgttcg
     4981 gatctttgct caggacgctc tgggtgctca gataatggtt atccggcagc agcaccgggc
     5041 catcgccaat cggggtgttc tgctgataat gatccgccag ctgcacgctg ccatcttcca
     5101 cgttgtggcg gattttaaag ttcgctttga tgccgttttt ctgtttatcc gcggtaatat
     5161 acacgttatg gctgttaaag ttgtattcca gtttatggcc caggatgttg ccatcttctt
     5221 taaaatcgat gcctttcagt tcaatgcggt tcaccagggt atcgccttca aatttcactt
     5281 ccgcacgggt tttataggtg ccatcatctt taaagctaat ggtgcgttcc tgcacatagc
     5341 cttccggcat cgcgctttta aaaaaatcat gctgtttcat atgatccgga taacgcgcaa
     5401 agcactgcac gccataggtc agggtggtca ccagggtcgg ccacggcacc ggcagtttgc
     5461 ccgtggtgca aatgaatttc agggtcagtt tgccgttggt cgcatcgcct tcgccttcgc
     5521 cacgcacgct aaatttatgg ccgttcacat cgccatccag ttccaccaga atcggcacca
     5581 caccggtaaa cagttcttcg cctttgcgca tgtgtttcaa tctcgattaa caggtaacgc
     5641 aaaattgctg tgtacgtcag attcatttta taacgcgaag ttccggaata aagtttctgg
     5701 aaaaagtaaa aatttattgt ccatctttac aaaagtccat ttactgttcg accgccatca
     5761 aacgccgata ttcatcccat gcgtaaaggt cagtcatccc gctgatataa tcctgaatca
     5821 accgacagcg ataataatat tccagtaccg ggtactccgc cgaatccgtg ggtaatttac
     5881 tgacgacttc cacgtaggcc aaacgatggc gggccggcat caccggcgcc acaggtgcgg
     5941 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc
     6001 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg
     6061 gcgccatctc ctttgctttt aagacccact ttcacattta agttgttttt ctaatccgca
     6121 tatgatcaat tcaaggccga ataagaaggc tggctctgca ccttggtgat caaataattc
     6181 gatagcttgt cgtaataatg gcggcatact atcagtagta ggtgtttccc tttcttcttt
     6241 agcgacttga tgctcttgat cttccaatac gcaacctaaa gtaaaatgcc ccacagcgct
     6301 gagtgcatat aatgcattct ctagtgaaaa accttgttgg cataaaaagg ctaattgatt
     6361 ttcgagagtt tcatactgtt tttctgtagg ccgtgtacct aaatgtactt ttgctccatc
     6421 gcgatgactt agtaaagcac atctaaaact tttagcgtta ttacgtaaaa aatcttgcca
     6481 gcttttccct tctaaagggc aaaagtgagt atggtgccta tctaacatct caatggctaa
     6541 ggcgtcgagc aaagcccgct tattttttac atgccaatac aatgtaggct gctctacacc
     6601 tagcctctgg gcgagtttac gggttgttaa accttcgatt ccgacctcat taagcagctc
     6661 taatgcgctg ctaatcactt tacttttatc taatctagac atcattaatt cctaattttt
//
