LOCUS       sequence-12924-e        7061 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 7061)
  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 7061)
  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: pBB535
FEATURES             Location/Qualifiers
     source          1..7061
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    16..32
                     /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             234..263
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(432..1514)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1476..1495
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1515..1592)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     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)."
     protein_bind    1695..1711
                     /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             1729..1740
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      4927..5013
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(5048..5065)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5182..5727)
                     /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(5262..5281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(5841..5935)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(5979..6770)
                     /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 gacgtctgtg tggaattgtg agcggataac aaattcacac agggccctcg gacaccgagg
       61 agaatgtcaa gaggcgaaca cacaacgtct tggagcgcca gaggaggaac gagctaaaac
      121 ggagcttttt tgccctgcgt gaccagatcc cggagttgga aaacaatgaa aaggccccca
      181 aggtagttat ccttaaaaaa gccacagcat acatcctgtc cgtccaagca gaggagcaaa
      241 agctcatttc tgaagaggac ttgttgcgga aacgacgaga acagttgaaa cacaaacttg
      301 aacagctacg gaactctttg tgcgtaagga aaagtaagga aaacgattcc ttctaacaga
      361 aatgtcctga gcaatcacct atgaactgtc gactcgacaa ttcgcgctaa cttacattaa
      421 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
      481 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt
      541 ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc
      601 agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac
      661 ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca
      721 ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg
      781 gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa
      841 ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg
      901 agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct
      961 aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg
     1021 tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac
     1081 gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag
     1141 ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct
     1201 tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga
     1261 gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg
     1321 ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc
     1381 agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg
     1441 ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac
     1501 gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata
     1561 ccgcgaaagg ttttgcacca ttcgatggtg tcaacgtaaa tgcatgccgc ttcgccttcg
     1621 cgcgcgaatt gtcgagaaaa tttatcaaaa agagtgttga cttgtgagcg gataacaatg
     1681 atacttagat tcaattgtga gcggataaca atttcacaca gaattcataa agaggagaaa
     1741 ggatccatgg gtaaaataat tggtatcgac ctgggtacta ccaactcttg tgtagcgatt
     1801 atggatggca ccactcctcg cgtgctggag aacgccgaag gcgatcgcac cacgccttct
     1861 atcattgcct atacccagga tggtgaaact ctagttggtc agccggctaa acgtcaggca
     1921 gtgacgaacc cgcaaaacac tctgtttgcg attaaacgcc tgattggtcg ccgcttccag
     1981 gacgaagaag tacagcgtga tgtttccatc atgccgttca aaattattgc tgctgataac
     2041 ggcgacgcat gggtcgaagt taaaggccag aaaatggcac cgccgcagat ttctgctgaa
     2101 gtgctgaaaa aaatgaagaa aaccgctgaa gattacctgg gtgaaccggt aactgaagct
     2161 gttatcaccg taccggcata ctttaacgat gctcagcgtc aggcaaccaa agacgcaggc
     2221 cgtatcgctg gtctggaagt aaaacgtatc atcaacgaac cgaccgcagc tgcgctggct
     2281 tacggtctgg acaaaggcac tggcaaccgt actatcgcgg tttatgacct gggtggtggt
     2341 actttcgata tttctattat cgaaatcgac gaagttgacg gcgaaaaaac cttcgaagtt
     2401 ctggcaacca acggtgatac ccacctgggg ggtgaagact tcgacagccg tctgatcaac
     2461 tatctggttg aagaattcaa gaaagatcag ggcattgacc tgcgcaacga tccgctggca
     2521 atgcagcgcc tgaaagaagc ggcagaaaaa gcgaaaatcg aactgtcttc cgctcagcag
     2581 accgacgtta acctgccata catcactgca gacgcgaccg gtccgaaaca catgaacatc
     2641 aaagtgactc gtgcgaaact ggaaagcctg gttgaagatc tggtaaaccg ttccattgag
     2701 ccgctgaaag ttgcactgca ggacgctggc ctgtccgtat ctgatatcga cgacgttatc
     2761 ctcgttggtg gtcagactcg tatgccaatg gttcagaaga aagttgctga gttctttggt
     2821 aaagagccgc gtaaagacgt taacccggac gaagctgtag caatcggtgc tgctgttcag
     2881 ggtggtgttc tgactggtga cgtaaaagac gtactgctgc tggacgttac cccgctgtct
     2941 ctgggtatcg aaaccatggg cggtgtgatg acgacgctga tcgcgaaaaa caccactatc
     3001 ccgaccaagc acagccaggt gttctctacc gctgaagaca accagtctgc ggtaaccatc
     3061 catgtgctgc agggtgaacg taaacgtgcg gctgataaca aatctctggg tcagttcaac
     3121 ctagatggta tcaacccggc accgcgcggc atgccgcaga tcgaagttac cttcgatatc
     3181 gatgctgacg gtatcctgca cgtttccgcg aaagataaaa acagcggtaa agagcagaag
     3241 atcaccatca aggcttcttc tggtctgaac gaagatgaaa tccagaaaat ggtacgcgac
     3301 gcagaagcta acgccgaagc tgaccgtaag tttgaagagc tggtacagac tcgcaaccag
     3361 ggcgaccatc tgctgcacag cacccgtaag caggttgaag aagcaggcga caaactgccg
     3421 gctgacgaca aaactgctat cgagtctgcg ctgactgcac tggaaactgc tctgaaaggt
     3481 gaagacaaag ccgctatcga agcgaaaatg caggaactgg cacaggtttc ccagaaactg
     3541 atggaaatcg cccagcagca acatgcccag cagcagactg ccggtgctga tgcttctgca
     3601 aacaacgcga aagatgacga tgttgtcgac gctgaatttg aagaagtcaa agacaaaaaa
     3661 taatcgccct ataaacgggt aattatactg acacgggcga aggggaattt cctctccgcc
     3721 cgtgcattca tctaggggca atttaaaaaa gatggctaag caagattatt acgagatttt
     3781 aggcgtttcc aaaacagcgg aagagcgtga aatcagaaag gcctacaaac gcctggccat
     3841 gaaataccac ccggaccgta accagggtga caaagaggcc gaggcgaaat ttaaagagat
     3901 caaggaagct tatgaagttc tgaccgactc gcaaaaacgt gcggcatacg atcagtatgg
     3961 tcatgctgcg tttgagcaag gtggcatggg cggcggcggt tttggcggcg gcgcagactt
     4021 cagcgatatt tttggtgacg ttttcggcga tatttttggc ggcggacgtg gtcgtcaacg
     4081 tgcggcgcgc ggtgctgatt tacgctataa catggagctc accctcgaag aagctgtacg
     4141 tggcgtgacc aaagagatcc gcattccgac tctggaagag tgtgacgttt gccacggtag
     4201 cggtgcaaaa ccaggtacac agccgcagac ttgtccgacc tgtcatggtt ctggtcaggt
     4261 gcagatgcgc cagggattct tcgctgtaca gcagacctgt ccacactgtc agggccgcgg
     4321 tacgctgatc aaagatccgt gcaacaaatg tcatggtcat ggtcgtgttg agcgcagcaa
     4381 aacgctgtcc gttaaaatcc cggcaggggt ggacactgga gaccgcatcc gtcttgcggg
     4441 cgaaggtgaa gcgggcgagc atggcgcacc ggcaggcgat ctgtacgttc aggttcaggt
     4501 taaacagcac ccgattttcg agcgtgaagg caacaacctg tattgcgaag tcccgatcaa
     4561 cttcgctatg gcggcgctgg gtggcgaaat cgaagtaccg acccttgatg gtcgcgtcaa
     4621 actgaaagtg cctggcgaaa cccagaccgg taagctattc cgtatgcgcg gtaaaggcgt
     4681 caagtctgtc cgcggtggcg cacagggtga tttgctgtgc cgcgttgtcg tcgaaacacc
     4741 ggtaggcctg aacgaaaggc agaaacagct gctgcaagag ctgcaagaaa gcttcggtgg
     4801 cccaaccggc gagcacaaca gcccgcgctc aaagagcttc tttgatggtg tgaagaagtt
     4861 ttttgacgac ctgacccgct aagatgtcga cctgcagcca agcttaatta gctgatctag
     4921 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     4981 ttgtcggtga acgctctcct gagtaggaca aatccgccgc cctagaccta gggatatatt
     5041 ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga gcggaaatgg
     5101 cttacgaacg gggcggagat ttcctggaag atgccaggaa gatacttaac agggaagtga
     5161 gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat
     5221 ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5281 ccctggcggc tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc
     5341 gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc
     5401 gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg
     5461 gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca
     5521 ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa
     5581 aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta
     5641 gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag
     5701 attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat aaaatatttc
     5761 tagatttcag tgcaatttat ctcttcaaat gtagcacctg aagtcagccc catacgatat
     5821 aagttgttac tagtgcttgg attctcacca ataaaaaacg cccggcggca accgagcgtt
     5881 ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag gagtccaagc
     5941 gagctcgata tccgtcggct tgaacgaatt gttagacatt atttgccgac taccttggtg
     6001 atctcgcctt tcacgtagtg gacaaattct tccaactgat ctgcgcgcga ggccaagcga
     6061 tcttcttctt gtccaagata agcctgtcta gcttcaagta tgacgggctg atactgggcc
     6121 ggcaggcgct ccattgccca gtcggcagcg acatccttcg gcgcgatttt gccggttact
     6181 gcgctgtacc aaatgcggga caacgtaagc actacatttc gctcatcgcc agcccagtcg
     6241 ggcggcgagt tccatagcgt taaggtttca tttagcgcct caaatagatc ctgttcagga
     6301 accggatcaa agagttcctc cgccgctgga cctaccaagg caacgctatg ttctcttgct
     6361 tttgtcagca agatagccag atcaatgtcg atcgtggctg gctcgaagat acctgcaaga
     6421 atgtcattgc gctgccattc tccaaattgc agttcgcgct tagctggata acgccacgga
     6481 atgatgtcgt cgtgcacaac aatggtgact tctacagcgc ggagaatctc gctctctcca
     6541 ggggaagccg aagtttccaa aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc
     6601 cttacggtca ccgtaaccag caaatcaata tcactgtgtg gcttcaggcc gccatccact
     6661 gcggagccgt acaaatgtac ggccagcaac gtcggttcga gatggcgctc gatgacgcca
     6721 actacctctg atagttgagt cgatacttcg gcgatcaccg cttccctcat gatgtttaac
     6781 tttgttttag ggcgactgcc ctgctgcgta acatcgttgc tgctccataa catcaaacat
     6841 cgacccacgg cgtaacgcgc ttgctgcttg gatgcccgag gcatagactg taccccaaaa
     6901 aaacagtcat aacaagccat gaaaaccgcc actgcgccgt taccaccgct gcgttcggtc
     6961 aaggttctgg accagttgcg tgagcgcata cgctacttgc attacagctt acgaaccgaa
     7021 caggcttatg tccactgggt tcgtgccttc atccggatat c
//
