LOCUS       sequence-258714-        3977 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for PURE system components.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3977)
  AUTHORS   Shimizu Y, Inoue A, Tomari Y, Suzuki T, Yokogawa T, Nishikawa K, 
            Ueda T
  TITLE     Cell-free translation reconstituted with purified components.
  JOURNAL   Nat Biotechnol. 2001 Aug;19(8):751-5. doi: 10.1038/90802.
  PUBMED    11479568
REFERENCE   2  (bases 1 to 3977)
  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"; doi: "10"; journalName: 
            "Nat Biotechnol"; date: "2001-08"; volume: "19"; issue: "8"; pages: 
            "751-5"
COMMENT     Sequence Label: pQE60-RRF-His
FEATURES             Location/Qualifiers
     source          1..3977
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(535..1326,1327..1395))
                     /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     1158..1177
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1396..1500)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1568..1586
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1610..1624,1625..1630,1631..1647,1648..1654)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    1630..1646
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    1662..1678
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             2280..2297
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2320..2414
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2524..2543)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3186..3272
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3319..3341)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3427..3567
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3482..3501
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3582..3599)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(3753..3977,1..364))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3833..3852)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
       61 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      121 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      181 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
      241 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      301 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      361 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
      421 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
      481 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
      541 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
      601 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
      661 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
      721 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
      781 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
      841 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
      901 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
      961 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1021 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1081 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1141 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1201 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1261 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1321 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     1381 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     1441 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     1501 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     1561 ctataaaaat aggcgtatca cgaggccctt tcgtcttcac ctcgagaaat cataaaaaat
     1621 ttatttgctt tgtgagcgga taacaattat aatagattca attgtgagcg gataacaatt
     1681 tcacacagaa ttcgttttca aggattcgta acatgattag cgatatcaga aaagatgctg
     1741 aagtacgcat ggacaaatgc gtagaagcgt tcaaaaccca aatcagcaaa atacgcacgg
     1801 gtcgtgcttc tcccagcctg ctggatggca ttgtcgtgga atattacggc acgccgacgc
     1861 cgctgcgtca gctggcaagc gtaacggtag aagattcccg tacactgaaa atcaacgtgt
     1921 ttgatcgttc aatgtctccg gccgttgaaa aagcgattat ggcgtccgat cttggcctga
     1981 acccgaactc tgcgggtagc gacatccgtg ttccgctgcc gccgctgacg gaagaacgtc
     2041 gtaaagatct gaccaaaatc gttcgtggtg aagcagaaca agcgcgtgtt gcagtacgta
     2101 acgtgcgtcg tgacgcgaac gacaaagtga aagcactgtt gaaagataaa gagatcagcg
     2161 aagacgacga tcgccgttct caggacgatg tacagaaact gactgatgct gcaatcaaga
     2221 aaattgaagc ggcgctggca gacaaagaag cagaactgat gcagttcgga tccagatctc
     2281 atcaccatca ccatcactaa gcttaattag ctgagcttgg actcctgttg atagatccag
     2341 taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt
     2401 ttattggtga gaatccaagc tagcttggcg agattttcag gagctaagga agctaaaatg
     2461 gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat
     2521 tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt
     2581 acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac
     2641 attcttgccc gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa agacggtgag
     2701 ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg
     2761 ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca catatattcg
     2821 caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat
     2881 atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc
     2941 aatatggaca acttcttcgc ccccgttttc accatgcatg ggcaaatatt atacgcaagg
     3001 cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca
     3061 tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta
     3121 atttttttaa ggcagttatt ggtgccctta aacgcctggg gtaatgactc tctagcttga
     3181 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     3241 tgtcggtgaa cgctctcctg agtaggacaa atccgccctc tagagctgcc tcgcgcgttt
     3301 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     3361 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     3421 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat
     3481 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     3541 tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg
     3601 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3661 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3721 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3781 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3841 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3901 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3961 aggtatctca gttcggt
//
