LOCUS       sequence-258688-        4596 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 4596)
  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 4596)
  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-EFTu-His
FEATURES             Location/Qualifiers
     source          1..4596
                     /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             2621..2638
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2661..2755
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2865..2884)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3527..3613
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3660..3682)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3768..3908
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3823..3842
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3923..3940)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(4094..4596,1..86))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4174..4193)
                     /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 aattctctta gagggacaat cgatgtctaa
     1441 agaaaagttt gaacgtacaa aaccgcacgt taacgtcggt actatcggcc acgttgacca
     1501 tggtaaaaca acgctgaccg ctgcaatcac taccgtactg gctaaaacct acggcggtgc
     1561 tgctcgcgca ttcgaccaga tcgataacgc gccggaagaa aaagctcgtg gtatcaccat
     1621 caacacttct cacgttgaat acgacacccc gacccgtcac tacgcacacg tagactgccc
     1681 ggggcacgcc gactatgtta aaaacatgat caccggtgct gcgcagatgg acggcgcgat
     1741 cctggtagtt gctgcgactg acggcccgat gccgcagact cgtgagcaca tcctgctggg
     1801 tcgtcaggta ggcgttccgt acatcatcgt gttcctgaac aaatgcgaca tggttgatga
     1861 cgaagagctg ctggaactgg ttgaaatgga agttcgtgaa cttctgtctc agtacgactt
     1921 cccgggcgac gacactccga tcgttcgtgg ttctgctctg aaagcgctgg aaggcgacgc
     1981 agagtgggaa gcgaaaatcc tggaactggc tggcttcctg gattcttaca ttccggaacc
     2041 agagcgtgcg attgacaagc cgttcctgct gccgatcgaa gacgtattct ccatctccgg
     2101 tcgtggtacc gttgttaccg gtcgtgtaga acgcggtatc atcaaagttg gtgaagaagt
     2161 tgaaatcgtt ggtatcaaag agactcagaa gtctacctgt actggcgttg aaatgttccg
     2221 caaactgctg gacgaaggcc gtgctggtga gaacgtaggt gttctgctgc gtggtatcaa
     2281 acgtgaagaa atcgaacgtg gtcaggtact ggctaagccg ggcaccatca agccgcacac
     2341 caagttcgaa tctgaagtgt acattctgtc caaagatgaa ggcggccgtc atactccgtt
     2401 cttcaaaggc taccgtccgc agttctactt ccgtactact gacgtgactg gtaccatcga
     2461 actgccggaa ggcgtagaga tggtaatgcc gggcgacaac atcaaaatgg ttgttaccct
     2521 gatccacccg atcgcgatgg acgacggtct gcgtttcgca atccgtgaag gcggccgtac
     2581 cgttggcgcg ggcgttgtag caaaagttct gggcagatct catcaccatc accatcacta
     2641 agcttaatta gctgagcttg gactcctgtt gatagatcca gtaatgacct cagaactcca
     2701 tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag
     2761 ctagcttggc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
     2821 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
     2881 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
     2941 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
     3001 atgctcatcc ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg
     3061 ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg
     3121 aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg
     3181 gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca
     3241 atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg
     3301 cccccgtttt caccatgcat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg
     3361 ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat
     3421 gaattacaac agtactgcga tgagtggcag ggcggggcgt aattttttta aggcagttat
     3481 tggtgccctt aaacgcctgg ggtaatgact ctctagcttg aggcatcaaa taaaacgaaa
     3541 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     3601 gagtaggaca aatccgccct ctagagctgc ctcgcgcgtt tcggtgatga cggtgaaaac
     3661 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     3721 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc
     3781 cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg
     3841 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     3901 gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3961 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4021 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4081 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4141 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4201 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4261 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4321 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4381 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4441 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4501 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     4561 tgaagccagt taccttcgga aaaagagttg gtagct
//
