LOCUS       sequence-12923-e        4089 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 4089)
  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 4089)
  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: pBB530
FEATURES             Location/Qualifiers
     source          1..4089
                     /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)."
     protein_bind    49..65
                     /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             84..95
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      743..837
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      853..939
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1477..2427)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2475..2697
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3189..3283)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3304..3963)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3878..3897
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3964..4066)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 gacgtctgtg tggaattgtg agcggataac aatgatactt agattcaatt gtgagcggat
       61 aacaatttca cacagaattc attaaagagg agaaattaac tatgagtagt aaagaacaga
      121 aaacgcctga ggggcaagcc ccggaagaaa ttatcatgga tcagcacgaa gagattgagg
      181 cagttgagcc agaagcttct gctgagcagg tggatccgcg cgatgaaaaa gttgcgaatc
      241 tcgaagctca gctggctgaa gcccagaccc gtgaacgtga cggcattttg cgtgtaaaag
      301 ccgaaatgga aaacctgcgt cgtcgtactg aactggatat tgaaaaagcc cacaaattcg
      361 cgctggagaa attcatcaac gaattgctgc cggtgattga tagcctggat cgtgcgctgg
      421 aagtggctga taaagctaac ccggatatgt ctgcgatggt tgaaggcatt gagctgacgc
      481 tgaagtcgat gctggatgtt gtgcgtaagt ttggcgttga agtgatcgcc gaaactaacg
      541 tcccactgga cccgaatgtg catcaggcca tcgcaatggt ggaatctgat gacgttgcgc
      601 caggtaacgt actgggcatt atgcagaagg gttatacgct gaatggtcgt acgattcgtg
      661 cggcgatggt tactgtagcg aaagcaaaag cttaatttct gctttcgtaa taactgcagc
      721 caagcttaat tagctgagct tggactcctg ttgatagatc cagtaatgac ctcagaactc
      781 catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg tgagaatcca
      841 agctagaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
      901 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgcccta gacctagggt
      961 acgggttttg ctgcccgcaa acgggctgtt ctggtgttgc tagtttgtta tcagaatcgc
     1021 agatccggct tcagccggtt tgccggctga aagcgctatt tcttccagaa ttgccatgat
     1081 tttttcccca cgggaggcgt cactggctcc cgtgttgtcg gcagctttga ttcgataagc
     1141 agcatcgcct gtttcaggct gtctatgtgt gactgttgag ctgtaacaag ttgtctcagg
     1201 tgttcaattt catgttctag ttgctttgtt ttactggttt cacctgttct attaggtgtt
     1261 acatgctgtt catctgttac attgtcgatc tgttcatggt gaacagcttt gaatgcacca
     1321 aaaactcgta aaagctctga tgtatctatc ttttttacac cgttttcatc tgtgcatatg
     1381 gacagttttc cctttgatat gtaacggtga acagttgttc tacttttgtt tgttagtctt
     1441 gatgcttcac tgatagatac aagagccata agaacctcag atccttccgt atttagccag
     1501 tatgttctct agtgtggttc gttgtttttg cgtgagccat gagaacgaac cattgagatc
     1561 atacttactt tgcatgtcac tcaaaaattt tgcctcaaaa ctggtgagct gaatttttgc
     1621 agttaaagca tcgtgtagtg tttttcttag tccgttatgt aggtaggaat ctgatgtaat
     1681 ggttgttggt attttgtcac cattcatttt tatctggttg ttctcaagtt cggttacgag
     1741 atccatttgt ctatctagtt caacttggaa aatcaacgta tcagtcgggc ggcctcgctt
     1801 atcaaccacc aatttcatat tgctgtaagt gtttaaatct ttacttattg gtttcaaaac
     1861 ccattggtta agccttttaa actcatggta gttattttca agcattaaca tgaacttaaa
     1921 ttcatcaagg ctaatctcta tatttgcctt gtgagttttc ttttgtgtta gttcttttaa
     1981 taaccactca taaatcctca tagagtattt gttttcaaaa gacttaacat gttccagatt
     2041 atattttatg aattttttta actggaaaag ataaggcaat atctcttcac taaaaactaa
     2101 ttctaatttt tcgcttgaga acttggcata gtttgtccac tggaaaatct caaagccttt
     2161 aaccaaagga ttcctgattt ccacagttct cgtcatcagc tctctggttg ctttagctaa
     2221 tacaccataa gcattttccc tactgatgtt catcatctga gcgtattggt tataagtgaa
     2281 cgataccgtc cgttctttcc ttgtagggtt ttcaatcgtg gggttgagta gtgccacaca
     2341 gcataaaatt agcttggttt catgctccgt taagtcatag cgactaatcg ctagttcatt
     2401 tgctttgaaa acaactaatt cagacataca tctcaattgg tctaggtgat tttaatcact
     2461 ataccaattg agatgggcta gtcaatgata attactagtc cttttcccgg gagatctggg
     2521 tatctgtaaa ttctgctaga cctttgctgg aaaacttgta aattctgcta gaccctctgt
     2581 aaattccgct agacctttgt gtgttttttt tgtttatatt caagtggtta taatttatag
     2641 aataaagaaa gaataaaaaa agataaaaag aatagatccc agccctgtgt ataactcact
     2701 actttagtca gttccgcagt attacaaaag gatgtcgcaa acgctgtttg ctcctctaca
     2761 aaacagacct taaaacccta aaggcttaag tagcaccctc gcaagctcgg gcaaatcgct
     2821 gaatattcct tttgtctccg accatcaggc acctgagtcg ctgtcttttt cgtgacattc
     2881 agttcgctgc gctcacggct ctggcagtga atgggggtaa atggcactac aggcgccttt
     2941 tatggattca tgcaaggaaa ctacccataa tacaagaaaa gcccgtcacg ggcttctcag
     3001 ggcgttttat ggcgggtctg ctatgtggtg ctatctgact ttttgctgtt cagcagttcc
     3061 tgccctctga ttttccagtc tgaccacttc ggattatccc gtgacaggtc attcagactg
     3121 gctaatgcac ccagtaaggc agcggtatca tcaacaggct tacccgtctt actgtcccta
     3181 gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc
     3241 agatggagtt ctgaggtcat tactggatct atcaacagga gtccaagcga gctcgatatc
     3301 aaattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc
     3361 gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt
     3421 gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt
     3481 ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat
     3541 attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg
     3601 cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa
     3661 cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag
     3721 ctcaccgtct ttcattgcca tacggaattc cggatgagca ttcatcaggc gggcaagaat
     3781 gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt
     3841 aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa
     3901 atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc
     3961 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     4021 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt ctcattttcg
     4081 ccagatatc
//
