LOCUS       sequence-258689-        4263 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 4263)
  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 4263)
  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-EFTs-His
FEATURES             Location/Qualifiers
     source          1..4263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(257..1048,1049..1117))
                     /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     880..899
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1118..1222)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1290..1308
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1332..1346,1347..1352,1353..1369,1370..1376)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    1352..1368
                     /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    1384..1400
                     /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             2288..2305
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2328..2422
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2532..2551)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3194..3280
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3327..3349)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3435..3575
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3490..3509
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3590..3607)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(3761..4263,1..86))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3841..3860)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
       61 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
      121 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
      181 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
      241 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
      301 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
      361 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
      421 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
      481 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
      541 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
      601 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
      661 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
      721 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
      781 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
      841 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
      901 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
      961 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     1021 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     1081 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     1141 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     1201 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     1261 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc
     1321 acctcgagaa atcataaaaa atttatttgc tttgtgagcg gataacaatt ataatagatt
     1381 caattgtgag cggataacaa tttcacacag aattgatctc cgaggatttt agatggctga
     1441 aattaccgca tccctggtaa aagagctgcg tgagcgtact ggcgcaggca tgatggattg
     1501 caaaaaagca ctgactgaag ctaacggcga catcgagctg gcaatcgaaa acatgcgtaa
     1561 gtccggtgct attaaagcag cgaaaaaagc aggcaacgtt gctgctgacg gcgtgatcaa
     1621 aaccaaaatc gacggcaact acggcatcat tctggaagtt aactgccaga ctgacttcgt
     1681 tgcaaaagac gctggtttcc aggcgttcgc agacaaagtt ctggacgcag ctgttgctgg
     1741 caaaatcact gacgttgaag ttctgaaagc acagttcgaa gaagaacgtg ttgcgctggt
     1801 agcgaaaatt ggtgaaaaca tcaacattcg ccgcgttgct gcgctggaag gcgacgttct
     1861 gggttcttat cagcacggtg cgcgtatcgg cgttctggtt gctgctaaag gcgctgacga
     1921 agagctggtt aaacacatcg ctatgcacgt tgctgcaagc aagccagaat tcatcaaacc
     1981 ggaagacgta tccgctgaag tggtagaaaa agaataccag gtacagctgg atatcgcgat
     2041 gcagtctggt aagccgaaag aaatcgcaga gaaaatggtt gaaggccgca tgaagaaatt
     2101 caccggcgaa gtttctctga ccggtcagcc gttcgttatg gaaccaagca aaactgttgg
     2161 tcagctgctg aaagagcata acgctgaagt gactggcttc atccgcttcg aagtgggtga
     2221 aggcatcgag aaagttgaga ctgactttgc agcagaagtt gctgcgatgt ccaagcagtc
     2281 tagatctcat caccatcacc atcactaagc ttaattagct gagcttggac tcctgttgat
     2341 agatccagta atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg
     2401 ggcgtttttt attggtgaga atccaagcta gcttggcgag attttcagga gctaaggaag
     2461 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     2521 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     2581 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     2641 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag
     2701 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
     2761 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     2821 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     2881 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     2941 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgcatggg caaatattat
     3001 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
     3061 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     3121 ggggcgtaat ttttttaagg cagttattgg tgcccttaaa cgcctggggt aatgactctc
     3181 tagcttgagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     3241 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccctcta gagctgcctc
     3301 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     3361 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     3421 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
     3481 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     3541 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg
     3601 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3661 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3721 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3781 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3841 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3901 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3961 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4021 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4081 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4141 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4201 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4261 gct
//
