LOCUS       sequence-240846-        5526 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 5526)
  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 5526)
  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-EFG-His
FEATURES             Location/Qualifiers
     source          1..5526
                     /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             3551..3568
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3591..3685
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3795..3814)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      4457..4543
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(4590..4612)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4698..4838
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4753..4772
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4853..4870)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5024..5526,1..86))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5104..5123)
                     /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 aattggataa acgaggaaac aaatggctcg
     1441 tacaacaccc atcgcacgct accgtaacat cggtatcagt gcgcacatcg acgccggtaa
     1501 aaccactact accgaacgta ttctgttcta caccggtgta aaccataaaa tcggtgaagt
     1561 tcatgacggc gctgcaacca tggactggat ggagcaggag caggaacgtg gtattaccat
     1621 cacttccgct gcgactactg cattctggtc tggtatggct aagcagtatg agccgcatcg
     1681 catcaacatc atcgacaccc cggggcacgt tgacttcaca atcgaagtag aacgttccat
     1741 gcgtgttctc gatggtgcgg taatggttta ctgcgcagtt ggtggtgttc agccgcagtc
     1801 tgaaaccgta tggcgtcagg caaacaaata taaagttccg cgcattgcgt tcgttaacaa
     1861 aatggaccgc atgggtgcga acttcctgaa agttgttaac cagatcaaaa cccgtctggg
     1921 cgcgaacccg gttccgctgc agctggcgat tggtgctgaa gaacatttca ccggtgttgt
     1981 tgacctggtg aaaatgaaag ctatcaactg gaacgacgct gaccagggcg taaccttcga
     2041 atacgaagat atcccggcag acatggttga actggctaac gaatggcacc agaacctgat
     2101 cgaatccgca gctgaagctt ctgaagagct gatggaaaaa tacctgggtg gtgaagaact
     2161 gactgaagca gaaatcaaag gtgctctgcg acagcgcgtt ctgaacaacg aaatcatcct
     2221 ggtaacctgt ggttctgcgt tcaagaacaa aggtgttcag gcgatgctgg atgcggtaat
     2281 tgattacctg ccatccccgg ttgacgtacc tgcgatcaac ggtatcctgg acgacggtaa
     2341 agacactccg gctgaacgtc acgcaagtga tgacgagccg ttctctgcac tggcgttcaa
     2401 aatcgctacc gacccgtttg ttggtaacct gaccttcttc cgtgtttact ccggtgtggt
     2461 taactctggt gataccgtac tgaactccgt gaaagctgca cgtgagcgtt tcggtcgtat
     2521 cgttcagatg cacgctaaca aacgtgaaga gatcaaagaa gttcgcgcgg gcgacatcgc
     2581 tgctgctatc ggtctgaaag acgtaaccac tggtgacacc ctgtgtgacc cggatgcgcc
     2641 gatcattctg gaacgtatgg aattccctga gccggtaatc tccatcgcag ttgaaccgaa
     2701 aaccaaagct gaccaggaaa aaatgggtct ggctctgggc cgtctggcta aagaagaccc
     2761 gtctttccgt gtatggactg acgaagaatc taaccagacc atcatcgcgg gtatgggcga
     2821 actgcacctc gacatcatcg ttgaccgtat gaagcgtgaa ttcaacgttg aagcgaacgt
     2881 aggtaaaccg caggttgctt accgtgaaac tatccgccag aaagttaccg atgttgaagg
     2941 taaacacgcg aaacagtctg gtggtcgtgg tcagtatggt catgttgtta tcgacatgta
     3001 cccgctggag ccgggttcaa acccgaaagg ctacgagttc atcaacgaca ttaaaggtgg
     3061 tgtaatccct ggcgaataca tcccggccgt tgataaaggt atccaggaac agctgaaagc
     3121 aggtccgctg gcaggctacc cggtagtaga catgggtatt cgtctgcact tcggttctta
     3181 ccatgacgtt gactcctctg aactggcgtt taaactggct gcttctatcg cctttaaaga
     3241 aggctttaag aaagcgaaac cagttctgct tgagccgatc atgaaggttg aagtagaaac
     3301 tccggaagag aacaccggtg acgttatcgg tgacttgagc cgtcgtcgtg gtatgctcaa
     3361 aggtcaggaa tctgaagtta ctggcgttaa gatccacgct gaagtaccgc tgtctgaaat
     3421 gttcggatac gcaactcagc tgcgttctct gaccaaaggt cgtgcatcat acactatgga
     3481 attcctgaag tatgatgaag cgccgagtaa cgttgctcag gccgtaattg aagcccgtgg
     3541 taaaagatct catcaccatc accatcacta agcttaatta gctgagcttg gactcctgtt
     3601 gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg
     3661 ccgggcgttt tttattggtg agaatccaag ctagcttggc gagattttca ggagctaagg
     3721 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
     3781 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
     3841 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
     3901 cctttattca cattcttgcc cgcctgatga atgctcatcc ggaatttcgt atggcaatga
     3961 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc
     4021 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac
     4081 acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt
     4141 ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt
     4201 taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgcat gggcaaatat
     4261 tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt
     4321 gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag
     4381 ggcggggcgt aattttttta aggcagttat tggtgccctt aaacgcctgg ggtaatgact
     4441 ctctagcttg aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     4501 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccct ctagagctgc
     4561 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     4621 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     4681 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact
     4741 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     4801 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca
     4861 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4921 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4981 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5041 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5101 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5161 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5221 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5281 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5341 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5401 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5461 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5521 gtagct
//
