LOCUS       sequence-389646-        6719 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 6719)
  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 6719)
  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: p5418
FEATURES             Location/Qualifiers
     source          1..6719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..24,25..29,30..35)
                     /label=tetA promoter
                     /note="contains a tet operator sequence that enables 
                     repression by TetR"
     protein_bind    6..24
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     RBS             66..77
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             81..758
                     /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(211..229)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(261..281)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     479..498
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     673..696
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             2130..2321
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2315..2337)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2423..2563
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2478..2497
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2578..2595)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2749..3337)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2829..2848)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3508..4299,4300..4368))
                     /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     4131..4150
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4369..4460)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(4479..4506)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4598..4684)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4837..4854
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(4896..5609)
                     /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(5948..5967)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(6077..6703)
                     /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     6608..6627
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
ORIGIN
        1 ttgacactct atcattgata gagttatttt accactccct atcagtgata agaaaagtga
       61 acgggaaaga ggagaaaagt atggcatcca ccgaggacgt catcaaggag ttcatgcgct
      121 tcaaagtgcg catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
      181 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc
      241 tgcccttcgc ctgggacatc ctgtcccccc agttccagta cggctccaag gtgtacgtga
      301 agcaccccgc cgacatcccc gactacaaga agctgtcctt ccccgagggc ttcaagtggg
      361 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
      421 aggacggctg cttcatctac aaggtgaagt tcatcggcgt gaacttcccc tccgacggcc
      481 ccgtaatgca gaagaagact atgggctggg agccctccac cgagcgcctg tacccccgcg
      541 acggcgtgct gaagggcgag atccacaagg ccctgaagct gaaggacggc ggccactacc
      601 tggtggagtt caagtccatc tacatggcca agaagcccgt gcaactgccc ggctactact
      661 acgtggactc caagctggac atcacctccc acaacgagga ctacaccatc gtggagcagt
      721 acgaacgcac cgagggccgc caccacctgt tcctgtagcc atggatcgat agctggtcga
      781 caaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg ctgcttccta
      841 atgcaggagt cgcataaggg agagcgtcga ccgatgccct tgagagcctt caacccagtc
      901 agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt
      961 atcatgcaac tcgtaggaca ggtgccggca gcgctctggg tcattttcgg cgaggaccgc
     1021 tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc
     1081 ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt
     1141 atcgccggca tggcggccga cgcgctgggc tacgtcttgc tggcgttcgc gacgcgaggc
     1201 tggatggcct tccccattat gattcttctc gcttccggcg gcatcgggat gcccgcgttg
     1261 caggccatgc tgtccaggca ggtagatgac gaccatcagg gacagcttca aggatcgctc
     1321 gcggctctta ccagcctaac ttcgatcatt ggaccgctga tcgtcacggc gatttatgcc
     1381 gcctcggcga gcacatggaa cgggttggca tggattgtag gcgccgccct ataccttgtc
     1441 tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca cctcgacctg aatggaagcc
     1501 ggcggcacct cgctaacgga ttcaccactc caagaattgg agccaatcaa ttcttgcgga
     1561 gaactgtgaa tgcgcaaacc aacccttggc agaacatatc catcgcgtcc gccatctcca
     1621 gcagccgcac gcggcgcatc tcgggcagcg ttgggtcctg gccacgggtg cgcatgatcg
     1681 tgctcctgtc gttgaggacc cggctaggct ggcggggttg ccttactggt tagcagaatg
     1741 aatcaccgat acgcgagcga acgtgaagcg actgctgctg caaaacgtct gcgacctgag
     1801 caacaacatg aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag
     1861 cgccctgcac cattatgttc cggatctgca tcgcaggatg ctgctggcta ccctgtggaa
     1921 cacctacatc tgtattaacg aagcgctggc attgaccctg agtgattttt ctctggtccc
     1981 gccgcatcca taccgccagt tgtttaccct cacaacgttc cagtaaccgg gcatgttcat
     2041 catcagtaac ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt acccccatga
     2101 acagaaatcc cccttacacg gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca
     2161 tggcccgctt tatcagaagc cagacattaa cgcttctgga gaaactcaac gagctggacg
     2221 cggatgaaca ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca
     2281 gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     2341 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     2401 cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt
     2461 atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     2521 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc
     2581 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     2641 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2701 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2761 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2821 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2881 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2941 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3001 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3061 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3121 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3181 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3241 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3301 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3361 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3421 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3481 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3541 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3601 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     3661 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3721 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3781 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3841 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3901 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3961 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4021 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4081 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc
     4141 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4201 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4261 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4321 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4381 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4441 tatttagaaa aataaacaaa agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg
     4501 ccttctgctt aatttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg
     4561 gccgttgctt cgcaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt
     4621 tcaccgacaa acaacagata aaacgaaagg cccagtcttt cgactgagcc tttcgtttta
     4681 tttgatgcct ggcagttccc tactctcgca tggggagacc ccacactacc atcggcgcta
     4741 cggcgtttca cttctgagtt cggcatgggg tcaggtggga ccaccgcgct actgccgcca
     4801 ggcaaattct gttttatcag accgcttctg cgttctgatt taatctgtat caggctgaaa
     4861 atcttctctc atccgccaaa acagaagttt tattatttat acagttcatc catgccatgc
     4921 gtaatacccg ccgcggtcac aaattccagc agcaccatat ggtcgcgttt ttcgttcgga
     4981 tctttgctca ggacgctctg ggtgctcaga taatggttat ccggcagcag caccgggcca
     5041 tcgccaatcg gggtgttctg ctgataatga tccgccagct gcacgctgcc atcttccacg
     5101 ttgtggcgga ttttaaagtt cgctttgatg ccgtttttct gtttatccgc ggtaatatac
     5161 acgttatggc tgttaaagtt gtattccagt ttatggccca ggatgttgcc atcttcttta
     5221 aaatcgatgc ctttcagttc aatgcggttc accagggtat cgccttcaaa tttcacttcc
     5281 gcacgggttt tataggtgcc atcatcttta aagctaatgg tgcgttcctg cacatagcct
     5341 tccggcatcg cgcttttaaa aaaatcatgc tgtttcatat gatccggata acgcgcaaag
     5401 cactgcacgc cataggtcag ggtggtcacc agggtcggcc acggcaccgg cagtttgccc
     5461 gtggtgcaaa tgaatttcag ggtcagtttg ccgttggtcg catcgccttc gccttcgcca
     5521 cgcacgctaa atttatggcc gttcacatcg ccatccagtt ccaccagaat cggcaccaca
     5581 ccggtaaaca gttcttcgcc tttgcgcatg gatgctgctc tcgtaccttg aatgaacgga
     5641 tagacaattc aaaacagatt gatgttattg aattttcagc ctggtaacaa tgataatagc
     5701 tgggacagga aagatagaga gagggcgcaa gcgatgcgcc cgataaattt cttaacggcg
     5761 tgcgattaat cgcgctttgc gccgctacgt aacaggccag acgcgccttc agaataatca
     5821 cgtggcattt gtactggcgc ctgatcgttg ctcgcctcag actccgccag acggttacgg
     5881 aacggattcg actccgcaga catttccggg gccggcatca ccggcgccac aggtgcggtt
     5941 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc
     6001 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc
     6061 gccatctcct ttgcttttaa gacccacttt cacatttaag ttgtttttct aatccgcata
     6121 tgatcaattc aaggccgaat aagaaggctg gctctgcacc ttggtgatca aataattcga
     6181 tagcttgtcg taataatggc ggcatactat cagtagtagg tgtttccctt tcttctttag
     6241 cgacttgatg ctcttgatct tccaatacgc aacctaaagt aaaatgcccc acagcgctga
     6301 gtgcatataa tgcattctct agtgaaaaac cttgttggca taaaaaggct aattgatttt
     6361 cgagagtttc atactgtttt tctgtaggcc gtgtacctaa atgtactttt gctccatcgc
     6421 gatgacttag taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa tcttgccagc
     6481 ttttcccttc taaagggcaa aagtgagtat ggtgcctatc taacatctca atggctaagg
     6541 cgtcgagcaa agcccgctta ttttttacat gccaatacaa tgtaggctgc tctacaccta
     6601 gcctctgggc gagtttacgg gttgttaaac cttcgattcc gacctcatta agcagctcta
     6661 atgcgctgct aatcacttta cttttatcta atctagacat cattaattcc taatttttg
//
