LOCUS       sequence-190340-        5126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for titrating synthesis of PdhR with doxycycline.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5126)
  AUTHORS   Sekar K, Rusconi R, Sauls JT, Fuhrer T, Noor E, Nguyen J, Fernandez 
            VI, Buffing MF, Berney M, Jun S, Stocker R, Sauer U
  TITLE     Synthesis and degradation of FtsZ quantitatively predict the first 
            cell division in starved bacteria.
  JOURNAL   Mol Syst Biol. 2018 Nov 5;14(11):e8623. doi: 10.15252/msb.20188623.
  PUBMED    30397005
REFERENCE   2  (bases 1 to 5126)
  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: "Mol Syst 
            Biol."; date: "2018-11-5"; volume: "14(11):e8623. doi"; pages: " 
            10.15252/msb.20188623"
COMMENT     Sequence Label: pPLtetO-PdhR
FEATURES             Location/Qualifiers
     source          1..5126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      99..185
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    347..365
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    372..390
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1496..1504
                     /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             1512..2135
                     /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="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(1585..1604)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2189..2236
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             join(2329..2397,2398..3189)
                     /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(2547..2566)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4341..4563
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     primer_bind     complement(5055..5072)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
ORIGIN
        1 ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt
       61 ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
      121 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
      181 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
      241 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatccgt caggatggcc
      301 tttttgcgtt tctacaaact ctgctagcaa gtaaggccga ctcgagtccc tatcagtgat
      361 agagattgac atccctatca gtgatagaga tactgagcac atcagcagga cgcatgaccg
      421 aattcattaa atttaacttt aagaaggaga tataatatgg cgtacagcaa aatccgccaa
      481 ccaaaactct ccgatgtgat tgagcagcaa ctggagtttt tgatcctcga aggcactctc
      541 cgcccgggcg aaaaactccc accggaacgc gaactggcaa aacagtttga cgtctcccgt
      601 ccctccttgc gtgaggcgat tcaacgtctc gaagcgaagg gcttgttgct tcgtcgccag
      661 ggtggcggca cttttgtcca gagcagccta tggcaaagct tcagcgatcc gctggtggag
      721 ctgctctccg accatcctga gtcacagtat gacttgctcg aaacacgaca cgccctggaa
      781 ggtatcgccg cttattacgc cgcgctgcgt agtaccgatg aagacaagga acgcatccgt
      841 gaactccacc acgccataga gctggcgcag cagtctggcg atctggacgc ggaatcaaac
      901 gccgtactcc agtatcagat tgccgtcacc gaagcggccc acaatgtggt tctgcttcat
      961 ctgctaaggt gtatggagcc gatgttggcc cagaatgtcc gccagaactt cgaattgctc
     1021 tattcgcgtc gcgagatgct gccgctggtg agtagtcacc gcacccgcat atttgaagcg
     1081 attatggccg gtaagccgga agaagcgcgc gaagcatcgc atcgccatct ggcctttatc
     1141 gaagaaattt tgctcgacag aagtcgtgaa gagagccgcc gtgagcgttc tctgcgtcgt
     1201 ctggagcaac gaaagaatta gcataaccct aatgagtgag ctaacttaca ttaattgcgt
     1261 tgcgccttaa ttaacggcac tcctcagcaa atataatgac cctcttgata acccaagagg
     1321 gcatttttta atgcccatgg cgtttaccac agctaacacc acgtcgtccc tatctgctgc
     1381 cctaggtcta tgagtggttg ctggataact ttacgggcat gcataaggct cgtaggctat
     1441 attcagggag accacaacgg tttccctcta caaataattt tgtttaactt tgaaataagg
     1501 aggtaataca aatgtctcgt ttagataaaa gtaaagtgat taacagcgca ttagagctgc
     1561 ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact cgcccagaag ctaggtgtag
     1621 agcagcctac attgtattgg catgtaaaaa ataagcgggc tttgctcgac gccttagcca
     1681 ttgagatgtt agataggcac catactcact tttgcccttt agaaggggaa agctggcaag
     1741 attttttacg taataacgct aaaagtttta gatgtgcttt actaagtcat cgcgatggag
     1801 caaaagtaca tttaggtaca cggcctacag aaaaacagta tgaaactctc gaaaatcaat
     1861 tagccttttt atgccaacaa ggtttttcac tagagaatgc attatatgca ctcagcgctg
     1921 tggggcattt tactttaggt tgcgtattgg aagatcaaga gcatcaagtc gctaaagaag
     1981 aaagggaaac acctactact gatagtatgc cgccattatt acgacaagct atcgaattat
     2041 ttgatcacca aggtgcagag ccagccttct tattcggcct tgaattgatc atatgcggat
     2101 tagaaaaaca acttaaatgt gaaagtgggt cttaataagc gactaaaaaa ttgaatgtag
     2161 ggaaccaaca tgccagttcg agcaataact agcataaccc cttggggcct ctaaacgggt
     2221 cttgaggggt tttttgctga aacctcaggc gagaagcaca cggtcacact tccgctcatg
     2281 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2341 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2401 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2461 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2521 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     2581 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2641 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2701 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     2761 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     2821 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     2881 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     2941 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3001 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3061 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3121 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3181 cattggtaac tgtcatacca agtttactca tatatacttt agattgattt aaaacttcat
     3241 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     3301 taacgtggag ttttcgttcc actgagcgtc agaccccgaa cctcagatcc ttccgtgttt
     3361 agccagtatg ttctctagtg tggttcgttg tttttgcgtg agccatgaga acgaaccatt
     3421 gagatcatac ttactttgca tgtcactcaa aaattttgcc tcaaaactgg tgagctgaat
     3481 ttttgcagtt aaagcatcgt gtagtgtttt tcttagtccg ttatgtaggt aggaatctga
     3541 tgtaatggtt gttggtattt tgtcaccatt catttttatc tggttgttct caagttcggt
     3601 tacgagatcc atttgtctat ctagttcaac ttggaaaatc aacgtatcag tcgggcggcc
     3661 tcgcttatca accaccaatt tcatattgct gtaagtgttt aaatctttac ttattggttt
     3721 caaaacccat tggttaagcc ttttaaactc atggtagtta ttttcaagca ttaacatgaa
     3781 cttaaattca tcaaggctaa tctctatatt tgccttgtga gttttctttt gtgttagttc
     3841 ttttaataac cactcataaa tcctcataga gtatttgttt tcaaaagact taacatgttc
     3901 cagattatat tttatgaatt tttttaactg gaaaagataa ggcaatatct cttcactaaa
     3961 aactaattct aatttttcgc ttgagaactt ggcatagttt gtccactgga aaatctcaaa
     4021 gcctttaacc aaaggattcc tgatttccac agttctcgtc atcagctctc tggttgcttt
     4081 agctaataca ccataagcat tttccctact gatgttcatc atctgagcgt attggttata
     4141 agtgaacgat accgtccgtt ctttccttgt agggttttca atcgtggggt tgagtagtgc
     4201 cacacagcat aaaattagct tggtttcatg ctccgttaag tcatagcgac taatcgctag
     4261 ttcatttgct ttgaaaacaa ctaatttaga catacatctc aattggtcta ggtgatttta
     4321 atcactatac caattgagat gggctagtca atgataatta ctagtccttt tcctttgagt
     4381 tgtgggtatc tgtaaattct gctagacctt tgctggaaaa cttgtaaatt ctgctagacc
     4441 ctctgtaaat tccgctagac ctttgtgtgt tttttttgtt tatattcaag tggttataat
     4501 ttatagaata aagaaagaat aaaaaaagat aaaaagaata gatcccagcc ctgtgtataa
     4561 ctcactactt tagtcagttc cgcagtatta caaaaggatg tcgcaaacgc tgtttgctcc
     4621 tctacaaaac agaccttaaa accctaaagg cttaagtagc accctcgcaa gctcgggcaa
     4681 atcgctgaat attccttttg tctccgacca tcaggcacct gagtcgctgt ctttttcgtg
     4741 acattcagtt cgctgcgctc acggctctgg cagtgaatgg gggtaaatgg cactacaggc
     4801 gccttttatg gattcatgca aggaaactac ccataataca agaaaagccc gtcacgggct
     4861 tctcagggcg ttttatggcg ggtctgctat gtggtgctat ctgacttttt gctgttcagc
     4921 agttcctgcc ctctgatttt ccagtctgac cacttcggat tatcccgtga caggtcattc
     4981 agactggcta atgcacccag taaggcagcg gtatcatcaa ctggctgttt tggcggatga
     5041 gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag
     5101 aatttgcctg gcggcagtag cgcggt
//
