LOCUS       sequence-142933-        4829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses sigma32 I54N mutant in E. coli to render a heat-shock like
            cellular folding environment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4829)
  AUTHORS   Zhang X, Liu Y, Genereux JC, Nolan C, Singh M, Kelly JW
  TITLE     The Heat-Shock Response Transcriptional Program Enables High-Yield 
            and High-Quality Recombinant Protein Production in Escherichia coli.
  JOURNAL   ACS Chem Biol. 2014 Jul 22.
  PUBMED    25051296
REFERENCE   2  (bases 1 to 4829)
  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: "ACS Chem 
            Biol. 2014 Jul 22."
COMMENT     Sequence Label: pBAD.Sigma32.I54N
FEATURES             Location/Qualifiers
     source          1..4829
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(34..53)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     194..216
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(372..1250)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1277..1561
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1484..1503
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(2491..2508)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2661..2747
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2839..2866
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2885..2976
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2977..3045,3046..3837)
                     /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     complement(3195..3214)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4008..4596
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4497..4516
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4750..4767
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cataggctgg ctggctctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
       61 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
      121 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
      181 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
      241 gcgaggcagc agatcaattc gcgcgcgaag gcgaagcggc atgcataatg tgcctgtcaa
      301 atggacgaag cagggattct gcaaacccta tgctactccg tcaagccgtc aattgtctga
      361 ttcgttacca attatgacaa cttgacggct acatcattca ctttttcttc acaaccggca
      421 cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa atagagttga
      481 tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt gctcaaaagc
      541 agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat ccctaactgc
      601 tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc gacgctggcg
      661 atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc gcgtacccga
      721 ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag taacaattgc
      781 tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc ggcgttaatg
      841 atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg aaagaacccc
      901 gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg cggacgaaag
      961 taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg atgaatctct
     1021 cctggcggga acagcaaaat atcacccggt cggcaaacaa attctcgtcc ctgatttttc
     1081 accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc cagcggtcgg
     1141 tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc caccagatgg
     1201 gcattaaacg agtatcccgg cagcagggga tcattttgcg cttcagccat acttttcata
     1261 ctcccgccat tcagagaaga aaccaattgt ccatattgca tcagacattg ccgtcactgc
     1321 gtcttttact ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa aagcattctg
     1381 taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta taatcacggc
     1441 agaaaagtcc acattgatta tttgcacggc gtcacacttt gctatgccat agcattttta
     1501 tccataagat tagcggatcc tacctgacgc tttttatcgc aactctctac tgtttctcca
     1561 tacccgtttt ttgggctaac aggaggaatt aaccatgact gacaaaatgc aaagtttagc
     1621 tttagcccca gttggcaacc tggattccta catccgggca gctaacgcgt ggccgatgtt
     1681 gtcggctgac gaggagcggg cgctggctga aaagctgcat taccatggcg atctggaagc
     1741 agctaaaacg ctgaacctgt ctcacctgcg gtttgttgtt catattgctc gtaattatgc
     1801 gggctatggc ctgccacagg cggatttgat tcaggaaggt aacatcggcc tgatgaaagc
     1861 agtgcgccgt ttcaacccgg aagtgggtgt gcgcctggtc tccttcgccg ttcactggat
     1921 caaagcagag atccacgaat acgttctgcg taactggcgt atcgtcaaag ttgcgaccac
     1981 caaagcgcag cgcaaactgt tcttcaacct gcgtaaaacc aagcagcgtc tgggctggtt
     2041 taaccaggat gaagtcgaaa tggtggcccg tgaactgggc gtaaccagca aagacgtacg
     2101 tgagatggaa tcacgtatgg cggcacagga catgaccttt gacctgtctt ccgacgacga
     2161 ttccgacagc cagccgatgg ctccggtgct ctatctgcag gataaatcat ctaactttgc
     2221 cgacggcatt gaagatgata actgggaaga gcaggcggca aaccgtctga ccgacgcgat
     2281 gcagggtctg gacgaacgca gccaggacat catccgtgcg cgctggctgg acgaagacaa
     2341 caagtccacg ttgcaggaac tggctgaccg ttacggcgtt tccgctgagc gtgtacgcca
     2401 gctggaaaag aacgcgatga aaaaattgcg tgctgccatt gaagcgtaat agaagcttgg
     2461 ctgttttggc ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag
     2521 cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat
     2581 gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag
     2641 agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     2701 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg
     2761 atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg
     2821 ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa
     2881 ctcttttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     2941 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     3001 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     3061 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     3121 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     3181 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     3241 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     3301 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     3361 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     3421 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     3481 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     3541 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     3601 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3661 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     3721 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3781 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3841 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3901 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3961 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4021 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4081 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4141 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4201 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4261 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4321 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4381 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4441 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4501 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4561 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4621 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4681 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4741 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     4801 cttacgcatc tgtgcggtat ttcacaccg
//
