LOCUS       sequence-190404-        5654 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Plasmid for titrating synthesis of ClpX with doxycycline.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5654)
  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 5654)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-ClpX
FEATURES             Location/Qualifiers
     source          1..5654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(59..127,128..919)
                     /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(277..296)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2071..2293
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     primer_bind     complement(2785..2802)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2955..3041
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    3203..3221
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3228..3246
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             4880..4888
                     /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             4896..5519
                     /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(4969..4988)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      5573..5620
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
       61 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
      121 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
      181 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
      241 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
      301 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      361 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      421 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
      481 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
      541 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
      601 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
      661 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
      721 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
      781 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
      841 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
      901 actgattaag cattggtaac tgtcatacca agtttactca tatatacttt agattgattt
      961 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1021 caaaatccct taacgtggag ttttcgttcc actgagcgtc agaccccgaa cctcagatcc
     1081 ttccgtgttt agccagtatg ttctctagtg tggttcgttg tttttgcgtg agccatgaga
     1141 acgaaccatt gagatcatac ttactttgca tgtcactcaa aaattttgcc tcaaaactgg
     1201 tgagctgaat ttttgcagtt aaagcatcgt gtagtgtttt tcttagtccg ttatgtaggt
     1261 aggaatctga tgtaatggtt gttggtattt tgtcaccatt catttttatc tggttgttct
     1321 caagttcggt tacgagatcc atttgtctat ctagttcaac ttggaaaatc aacgtatcag
     1381 tcgggcggcc tcgcttatca accaccaatt tcatattgct gtaagtgttt aaatctttac
     1441 ttattggttt caaaacccat tggttaagcc ttttaaactc atggtagtta ttttcaagca
     1501 ttaacatgaa cttaaattca tcaaggctaa tctctatatt tgccttgtga gttttctttt
     1561 gtgttagttc ttttaataac cactcataaa tcctcataga gtatttgttt tcaaaagact
     1621 taacatgttc cagattatat tttatgaatt tttttaactg gaaaagataa ggcaatatct
     1681 cttcactaaa aactaattct aatttttcgc ttgagaactt ggcatagttt gtccactgga
     1741 aaatctcaaa gcctttaacc aaaggattcc tgatttccac agttctcgtc atcagctctc
     1801 tggttgcttt agctaataca ccataagcat tttccctact gatgttcatc atctgagcgt
     1861 attggttata agtgaacgat accgtccgtt ctttccttgt agggttttca atcgtggggt
     1921 tgagtagtgc cacacagcat aaaattagct tggtttcatg ctccgttaag tcatagcgac
     1981 taatcgctag ttcatttgct ttgaaaacaa ctaatttaga catacatctc aattggtcta
     2041 ggtgatttta atcactatac caattgagat gggctagtca atgataatta ctagtccttt
     2101 tcctttgagt tgtgggtatc tgtaaattct gctagacctt tgctggaaaa cttgtaaatt
     2161 ctgctagacc ctctgtaaat tccgctagac ctttgtgtgt tttttttgtt tatattcaag
     2221 tggttataat ttatagaata aagaaagaat aaaaaaagat aaaaagaata gatcccagcc
     2281 ctgtgtataa ctcactactt tagtcagttc cgcagtatta caaaaggatg tcgcaaacgc
     2341 tgtttgctcc tctacaaaac agaccttaaa accctaaagg cttaagtagc accctcgcaa
     2401 gctcgggcaa atcgctgaat attccttttg tctccgacca tcaggcacct gagtcgctgt
     2461 ctttttcgtg acattcagtt cgctgcgctc acggctctgg cagtgaatgg gggtaaatgg
     2521 cactacaggc gccttttatg gattcatgca aggaaactac ccataataca agaaaagccc
     2581 gtcacgggct tctcagggcg ttttatggcg ggtctgctat gtggtgctat ctgacttttt
     2641 gctgttcagc agttcctgcc ctctgatttt ccagtctgac cacttcggat tatcccgtga
     2701 caggtcattc agactggcta atgcacccag taaggcagcg gtatcatcaa ctggctgttt
     2761 tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct
     2821 gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa
     2881 ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg
     2941 gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     3001 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
     3061 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc
     3121 atcaaattaa gcagaaggcc atccgtcagg atggcctttt tgcgtttcta caaactctgc
     3181 tagcaagtaa ggccgactcg agtccctatc agtgatagag attgacatcc ctatcagtga
     3241 tagagatact gagcacatca gcaggacgca tgaccgaatt cattaaattt aactttaaga
     3301 aggagatata atatgacaga taaacgcaaa gatggctcag gcaaattgct gtattgctct
     3361 ttttgcggca aaagccagca tgaagtgcgc aagctgattg ccggtccatc cgtgtatatc
     3421 tgcgacgaat gtgttgattt atgtaacgac atcattcgcg aagagattaa agaagttgca
     3481 ccgcatcgtg aacgcagtgc gctaccgacg ccgcatgaaa ttcgcaacca cctggacgat
     3541 tacgttatcg gccaggaaca ggcgaaaaaa gtgctggcgg tcgcggtata caaccattac
     3601 aaacgtctgc gcaacggcga taccagcaat ggcgtcgagt tgggcaaaag taacattctg
     3661 ctgatcggtc cgaccggttc cggtaaaacg ctgctggctg aaacgctggc gcgcctgctg
     3721 gatgttccgt tcaccatggc cgacgcgact acactgaccg aagccggtta tgtgggtgaa
     3781 gacgttgaaa acatcattca gaagctgttg cagaaatgcg actacgatgt ccagaaagca
     3841 cagcgtggta ttgtctacat cgatgaaatc gacaagattt ctcgtaagtc agacaacccg
     3901 tccattaccc gagacgtttc cggtgaaggc gtacagcagg cactgttgaa actgatcgaa
     3961 ggtacggtag ctgctgttcc accgcaaggt gggcgtaaac atccgcagca ggaattcttg
     4021 caggttgata cctctaagat cctgtttatt tgtggcggtg cgtttgccgg tctggataaa
     4081 gtgatttccc accgtgtaga aaccggctcc ggcattggtt ttggcgcgac ggtaaaagcg
     4141 aagtccgaca aagcaagcga aggcgagctg ctggcgcagg ttgaaccgga agatctgatc
     4201 aagtttggtc ttatccctga gtttattggt cgtctgccgg ttgtcgcaac gttgaatgaa
     4261 ctgagcgaag aagctctgat tcagatcctc aaagagccga aaaacgccct gaccaagcag
     4321 tatcaggcgc tgtttaatct ggaaggcgtg gatctggaat tccgtgacga ggcgctggat
     4381 gctatcgcta agaaagcgat ggcgcgtaaa accggtgccc gtggcctgcg ttccatcgta
     4441 gaagccgcac tgctcgatac catgtacgat ctgccgtcca tggaagacgt cgaaaaagtg
     4501 gttatcgacg agtcggtaat tgatggtcaa agcaaaccgt tgctgattta tggcaagccg
     4561 gaagcgcaac aggcatctgg tgaataagat gcatgccagt tctagcataa ccctaatgag
     4621 tgagctaact tacattaatt gcgttgcgcc ttaattaacg gcactcctca gcaaatataa
     4681 tgaccctctt gataacccaa gagggcattt tttaatgccc atggcgttta ccacagctaa
     4741 caccacgtcg tccctatctg ctgccctagg tctatgagtg gttgctggat aactttacgg
     4801 gcatgcataa ggctcgtagg ctatattcag ggagaccaca acggtttccc tctacaaata
     4861 attttgttta actttgaaat aaggaggtaa tacaaatgtc tcgtttagat aaaagtaaag
     4921 tgattaacag cgcattagag ctgcttaatg aggtcggaat cgaaggttta acaacccgta
     4981 aactcgccca gaagctaggt gtagagcagc ctacattgta ttggcatgta aaaaataagc
     5041 gggctttgct cgacgcctta gccattgaga tgttagatag gcaccatact cacttttgcc
     5101 ctttagaagg ggaaagctgg caagattttt tacgtaataa cgctaaaagt tttagatgtg
     5161 ctttactaag tcatcgcgat ggagcaaaag tacatttagg tacacggcct acagaaaaac
     5221 agtatgaaac tctcgaaaat caattagcct ttttatgcca acaaggtttt tcactagaga
     5281 atgcattata tgcactcagc gctgtggggc attttacttt aggttgcgta ttggaagatc
     5341 aagagcatca agtcgctaaa gaagaaaggg aaacacctac tactgatagt atgccgccat
     5401 tattacgaca agctatcgaa ttatttgatc accaaggtgc agagccagcc ttcttattcg
     5461 gccttgaatt gatcatatgc ggattagaaa aacaacttaa atgtgaaagt gggtcttaat
     5521 aagcgactaa aaaattgaat gtagggaacc aacatgccag ttcgagcaat aactagcata
     5581 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaacctc aggcgagaag
     5641 cacacggtca cact
//
