LOCUS       sequence-209089-        4440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4440)
  AUTHORS   de Marco A, Deuerling E, Mogk A, Tomoyasu T, Bukau B
  TITLE     Chaperone-based procedure to increase yields of soluble recombinant 
            proteins produced in E. coli.
  JOURNAL   BMC Biotechnol. 2007 Jun 12;7:32. doi: 10.1186/1472-6750-7-32.
  PUBMED    17565681
REFERENCE   2  (bases 1 to 4440)
  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: "BMC 
            Biotechnol."; date: "2007-06-12"; volume: "7:32. doi"; pages: " 
            10.1186/1472-6750-7-32"
COMMENT     Sequence Label: pBB541
FEATURES             Location/Qualifiers
     source          1..4440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     43..61
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    107..123
                     /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    140..156
                     /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)."
     RBS             175..186
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             238..291
                     /codon_start=1
                     /product="GroES chaperone mobile loop that interacts with 
                     GroEL"
                     /label=S loop
                     /translation="EVETKSAGGIVLTGSAAA"
     terminator      2196..2290
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      2306..2392
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2427..2444)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2561..3106)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2641..2660)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3220..3314)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3358..4149)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
       61 ccctttcgtc ttcacctcga gaaaatttat caaaaagagt gttgacttgt gagcggataa
      121 caatgatact tagattcaat tgtgagcgga taacaatttc acacagaatt cattaaagag
      181 gagaaattaa ccatgaatat tcgtccattg catgatcgcg tgatcgtcaa gcgtaaagaa
      241 gttgaaacta aatctgctgg cggcatcgtt ctgaccggct ctgcagcggc taaatccacc
      301 cgcggcgaag tgctggctgt cggcaatggc cgtatccttg aaaatggcga agtgaagccg
      361 ctggatgtga aagttggcga catcgttatt ttcaacgatg gctacggtgt gaaatctgag
      421 aagatcgaca atgaagaagt gttgatcatg tccgaaagcg acattctggc aattgttgaa
      481 gcgtaatccg cgcacgacac tgaacatacg aatttaagga ataaagataa tggcagctaa
      541 agacgtaaaa ttcggtaacg acgctcgtgt gaaaatgctg cgcggcgtaa acgtactggc
      601 agatgcagtg aaagttaccc tcggtccaaa aggccgtaac gtagttctgg ataaatcttt
      661 cggtgcaccg accatcacca aagatggtgt ttccgttgct cgtgaaatcg aactggaaga
      721 caagttcgaa aatatgggtg cgcagatggt gaaagaagtt gcctctaaag caaacgacgc
      781 tgcaggcgac ggtaccacca ctgcaaccgt actggctcag gctatcatca ctgaaggtct
      841 gaaagctgtt gctgcgggca tgaacccgat ggacctgaaa cgtggtatcg acaaagcggt
      901 taccgctgca gttgaagaac tgaaagcgct gtccgtacca tgctctgact ctaaagcgat
      961 tgctcaggtt ggtaccatct ccgctaactc cgacgaaacc gtaggtaaac tgatcgctga
     1021 agcgatggac aaagtcggta aagaaggcgt tatcaccgtt gaagacggta ccggtctgca
     1081 ggacgaactg gacgtggttg aaggtatgca gttcgaccgt ggctacctgt ctccttactt
     1141 catcaacaag ccggaaactg gcgcagtaga actggaaagc ccgttcatcc tgctggctga
     1201 caagaaaatc tccaacatcc gcgaaatgct gccggttctg gaagctgttg ccaaagcagg
     1261 caaaccgctg ctgatcatcg ctgaagatgt agaaggcgaa gcgctggcaa ctctggttgt
     1321 taacaccatg cgtggcatcg tgaaagtcgc tgcggttaaa gcaccgggct tcggcgatcg
     1381 tcgtaaagct atgctgcagg atatcgcaac cctgactggc ggtaccgtga tctctgaaga
     1441 gatcggtatg gagctggaaa aagcaaccct ggaagacctg ggtcaggcta aacgtgttgt
     1501 gatcaacaaa gacaccacca ctatcatcga tggcgtgggt gaagaagctg caatccaggg
     1561 ccgtgttgct cagatccgtc agcagattga agaagcaact tctgactacg accgtgaaaa
     1621 actgcaggaa cgcgtagcga aactggcagg cggcgttgca gttatcaaag tgggtgctgc
     1681 taccgaagtt gaaatgaaag agaaaaaagc acgcgttgaa gatgccctgc acgcgacccg
     1741 tgctgcggta gaagaaggcg tggttgctgg tggtggtgtt gcgctgatcc gcgtagcgtc
     1801 taaactggct gacctgcgtg gtcagaacga agaccagaac gtgggtatca aagttgcact
     1861 gcgtgcaatg gaagctccgc tgcgtcagat cgtattgaac tgcggcgaag aaccgtctgt
     1921 tgttgctaac accgttaaag gcggcgacgg caactacggt tacaacgcag caaccgaaga
     1981 atacggcaac atgatcgaca tgggtatcct ggatccaacc aaagtaactc gttctgctct
     2041 gcagtacgca gcttctgtgg ctggcctgat gatcaccacc gaatgcatgg ttaccgacct
     2101 gccgaaaaac gatgcagctg acttaggcgc tgctggcggt atgggcggca tgggtggcat
     2161 gggcggcatg atgtaagctt aattagctga gcttggactc ctgttgatag atccagtaat
     2221 gacctcagaa ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat
     2281 tggtgagaat ccaagctaga ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     2341 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccgcc
     2401 ctagacctag ggatatattc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga
     2461 ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt tcctggaaga tgccaggaag
     2521 atacttaaca gggaagtgag agggccgcgg caaagccgtt tttccatagg ctccgccccc
     2581 ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat
     2641 aaagatacca ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc
     2701 ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc
     2761 agttccgggt aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc
     2821 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa
     2881 gcaccactgg cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc
     2941 cggttaaggc taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc
     3001 tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt
     3061 ttcgttttca gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat
     3121 taatcagata aaatatttct agatttcagt gcaatttatc tcttcaaatg tagcacctga
     3181 agtcagcccc atacgatata agttgttact agtgcttgga ttctcaccaa taaaaaacgc
     3241 ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc
     3301 tatcaacagg agtccaagcg agctcgatat ccgtcggctt gaacgaattg ttagacatta
     3361 tttgccgact accttggtga tctcgccttt cacgtagtgg acaaattctt ccaactgatc
     3421 tgcgcgcgag gccaagcgat cttcttcttg tccaagataa gcctgtctag cttcaagtat
     3481 gacgggctga tactgggccg gcaggcgctc cattgcccag tcggcagcga catccttcgg
     3541 cgcgattttg ccggttactg cgctgtacca aatgcgggac aacgtaagca ctacatttcg
     3601 ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat ttagcgcctc
     3661 aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc gccgctggac ctaccaaggc
     3721 aacgctatgt tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga tcgtggctgg
     3781 ctcgaagata cctgcaagaa tgtcattgcg ctgccattct ccaaattgca gttcgcgctt
     3841 agctggataa cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt ctacagcgcg
     3901 gagaatctcg ctctctccag gggaagccga agtttccaaa aggtcgttga tcaaagctcg
     3961 ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat cactgtgtgg
     4021 cttcaggccg ccatccactg cggagccgta caaatgtacg gccagcaacg tcggttcgag
     4081 atggcgctcg atgacgccaa ctacctctga tagttgagtc gatacttcgg cgatcaccgc
     4141 ttccctcatg atgtttaact ttgttttagg gcgactgccc tgctgcgtaa catcgttgct
     4201 gctccataac atcaaacatc gacccacggc gtaacgcgct tgctgcttgg atgcccgagg
     4261 catagactgt accccaaaaa aacagtcata acaagccatg aaaaccgcca ctgcgccgtt
     4321 accaccgctg cgttcggtca aggttctgga ccagttgcgt gagcgcatac gctacttgca
     4381 ttacagctta cgaaccgaac aggcttatgt ccactgggtt cgtgccttca tccggatatc
//
