LOCUS       sequence-44491-e        7203 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7203)
  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 . 7():32.
  PUBMED    17565681
REFERENCE   2  (bases 1 to 7203)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "BMC 
            Biotechnol. 2007 . 7():32."
COMMENT     Sequence Label: pBB540
FEATURES             Location/Qualifiers
     source          1..7203
                     /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"
     protein_bind    423..439
                     /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    456..472
                     /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             491..502
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1150..1244
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      1260..1346
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    1395..1411
                     /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    1428..1444
                     /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             1463..1474
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             complement(4591..5541)
                     /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      5589..5811
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(6303..6397)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6418..7077)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6992..7011
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(7078..7180)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
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 gaactcttgt gcgtaaggaa aagtaaggaa aacgattcct tctaacagaa
      361 atgtcctgag caatcaccta tgaactgtcg actcgagaaa atttatcaaa aagagtgttg
      421 acttgtgagc ggataacaat gatacttaga ttcaattgtg agcggataac aatttcacac
      481 agaattcatt aaagaggaga aattaactat gagtagtaaa gaacagaaaa cgcctgaggg
      541 gcaagccccg gaagaaatta tcatggatca gcacgaagag attgaggcag ttgagccaga
      601 agcttctgct gagcaggtgg atccgcgcga tgaaaaagtt gcgaatctcg aagctcagct
      661 ggctgaagcc cagacccgtg aacgtgacgg cattttgcgt gtaaaagccg aaatggaaaa
      721 cctgcgtcgt cgtactgaac tggatattga aaaagcccac aaattcgcgc tggagaaatt
      781 catcaacgaa ttgctgccgg tgattgatag cctggatcgt gcgctggaag tggctgataa
      841 agctaacccg gatatgtctg cgatggttga aggcattgag ctgacgctga agtcgatgct
      901 ggatgttgtg cgtaagtttg gcgttgaagt gatcgccgaa actaacgtcc cactggaccc
      961 gaatgtgcat caggccatcg caatggtgga atctgatgac gttgcgccag gtaacgtact
     1021 gggcattatg cagaagggtt atacgctgaa tggtcgtacg attcgtgcgg cgatggttac
     1081 tgtagcgaaa gcaaaagctt aatttctgct ttcgtaataa ctgcagccaa gcttaattag
     1141 ctgagcttgg actcctgttg atagatccag taatgacctc agaactccat ctggatttgt
     1201 tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaagc tagaggcatc
     1261 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     1321 tgaacgctct cctgagtagg acaaatccgc cgccctagac ctagtcgaga aaatttatca
     1381 aaaagagtgt tgacttgtga gcggataaca atgatactta gattcaattg tgagcggata
     1441 acaatttcac acagaattca ttaaagagga gaaaggatct atgggaggag ttatgcgtct
     1501 ggatcgtctt actaataaat tccagcttgc tcttgccgat gcccaatcac ttgcactcgg
     1561 gcacgacaac caatttatcg aaccacttca tttaatgagc gccctgctga atcaggaagg
     1621 gggttcggtt agtcctttat taacatccgc tggcataaat gctggccagt tgcgcacaga
     1681 tatcaatcag gcattaaatc gtttaccgca ggttgaaggt actggtggtg atgtccagcc
     1741 atcacaggat ctggtgcgcg ttcttaatct ttgcgacaag ctggcgcaaa aacgtggtga
     1801 taactttatc tcgtcagaac tgttcgttct ggcggcactt gagtctcgcg gcacgctggc
     1861 cgacatcctg aaagcagcag gggcgaccac cgccaacatt actcaagcga ttgaacaaat
     1921 gcgtggaggt gaaagcgtga acgatcaagg tgctgaagac caacgtcagg ctttgaaaaa
     1981 atataccatc gaccttaccg aacgagccga acagggcaaa ctcgatccgg tgattggtcg
     2041 tgatgaagaa attcgccgta ccattcaggt gctgcaacgt cgtactaaaa ataacccggt
     2101 actgattggt gaacccggcg tcggtaaaac tgccatcgtt gaaggtctgg cgcagcgtat
     2161 tatcaacggc gaagtgccgg aagggttgaa aggccgccgg gtactggcgc tggatatggg
     2221 cgcgctggtg gctggggcga aatatcgcgg tgagtttgaa gaacgtttaa aaggcgtgct
     2281 taacgatctt gccaaacagg aaggcaacgt catcctattt atcgacgaat tacataccat
     2341 ggtcggcgcg ggtaaagccg atggcgcaat ggacgccgga aacatgctga aaccggcgct
     2401 ggcgcgtggt gaattgcact gcgtaggtgc cacgacgctt gacgaatatc gccagtacat
     2461 tgaaaaagat gctgcgctgg aacgtcgttt ccagaaagtg tttgttgccg agccttctgt
     2521 tgaagatacc attgcgattc tgcgtggcct gaaagaacgt tacgaattgc accaccatgt
     2581 gcaaattact gacccggcaa ttgttgcagc ggcgacgttg tctcatcgct acattgctga
     2641 ccgtcagctg ccggataaag ccatcgacct gatcgatgaa gcagcatcca gcattcgtat
     2701 gcagattgac tcaaaaccag aagaactcga ccgactcgat cgtcgtatca tccagctcaa
     2761 actggaacaa caggcgttaa tgaaagagtc tgatgaagcc agtaaaaaac gtctggatat
     2821 gctcaacgaa gaactgagcg acaaagaacg tcagtactcc gagttagaag aagagtggaa
     2881 agcagagaag gcatcgcttt ctggtacgca gaccattaaa gcggaactgg aacaggcgaa
     2941 aatcgctatt gaacaggctc gccgtgtggg ggacctggcg cggatgtctg aactgcaata
     3001 cggcaaaatc ccggaactgg aaaagcaact ggaagccgca acgcagctcg aaggcaaaac
     3061 tatgcgtctg ttgcgtaata aagtgaccga cgccgaaatt gctgaagtgc tggcgcgttg
     3121 gacggggatt ccggtttctc gcatgatgga aagcgagcgc gaaaaactgc tgcgtatgga
     3181 gcaagaactg caccatcgcg taattggtca gaacgaagcg gttgatgcgg tatctaacgc
     3241 tattcgtcgt agccgtgcgg ggctggcgga tccaaatcgc ccgattggtt cattcctgtt
     3301 cctcggccca actggtgtgg ggaaaacaga gctttgtaag gcgctggcga actttatgtt
     3361 tgatagcgac gaggcgatgg tccgtatcga tatgtccgag tttatggaga aacactcggt
     3421 gtctcgtttg gttggtgcgc ctccgggata tgtcggttat gaagaaggtg gctacctgac
     3481 cgaagcggtg cgtcgtcgtc cgtattccgt catcctgctg gatgaagtgg aaaaagcgca
     3541 tccggatgtc ttcaacattc tgttgcaggt actggatgat gggcgtctga ctgacgggca
     3601 agggagaacg gtcgacttcc gtaatacggt cgtcattatg acctctaacc tcggttccga
     3661 tctgattcag gaacgcttcg gtgaactgga ttatgcgcac atgaaagagc tggtgctcgg
     3721 tgtggtaagc cataacttcc gtccggaatt cattaaccgt atcgatgaag tggtggtctt
     3781 ccatccgctg ggtgaacagc acattgcctc gattgcgcag attcagttga aacgtctgta
     3841 caaacgtctg gaagaacgtg gttatgaaat ccacatttct gacgaggcgc tgaaactgct
     3901 gagcgagaac ggttacgatc cggtctatgg tgcacgtcct ctgaaacgtg caattcagca
     3961 gcagatcgaa aacccgctgg cacagcaaat actgtctggt gaattggttc cgggtaaagt
     4021 gattcgcctg gaagttaatg aagaccggat tgtcgccgtc cagtaatcta gggtacgggt
     4081 tttgctgccc gcaaacgggc tgttctggtg ttgctagttt gttatcagaa tcgcagatcc
     4141 ggcttcagcc ggtttgccgg ctgaaagcgc tatttcttcc agaattgcca tgattttttc
     4201 cccacgggag gcgtcactgg ctcccgtgtt gtcggcagct ttgattcgat aagcagcatc
     4261 gcctgtttca ggctgtctat gtgtgactgt tgagctgtaa caagttgtct caggtgttca
     4321 atttcatgtt ctagttgctt tgttttactg gtttcacctg ttctattagg tgttacatgc
     4381 tgttcatctg ttacattgtc gatctgttca tggtgaacag ctttgaatgc accaaaaact
     4441 cgtaaaagct ctgatgtatc tatctttttt acaccgtttt catctgtgca tatggacagt
     4501 tttccctttg atatgtaacg gtgaacagtt gttctacttt tgtttgttag tcttgatgct
     4561 tcactgatag atacaagagc cataagaacc tcagatcctt ccgtatttag ccagtatgtt
     4621 ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac gaaccattga gatcatactt
     4681 actttgcatg tcactcaaaa attttgcctc aaaactggtg agctgaattt ttgcagttaa
     4741 agcatcgtgt agtgtttttc ttagtccgtt atgtaggtag gaatctgatg taatggttgt
     4801 tggtattttg tcaccattca tttttatctg gttgttctca agttcggtta cgagatccat
     4861 ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc gggcggcctc gcttatcaac
     4921 caccaatttc atattgctgt aagtgtttaa atctttactt attggtttca aaacccattg
     4981 gttaagcctt ttaaactcat ggtagttatt ttcaagcatt aacatgaact taaattcatc
     5041 aaggctaatc tctatatttg ccttgtgagt tttcttttgt gttagttctt ttaataacca
     5101 ctcataaatc ctcatagagt atttgttttc aaaagactta acatgttcca gattatattt
     5161 tatgaatttt tttaactgga aaagataagg caatatctct tcactaaaaa ctaattctaa
     5221 tttttcgctt gagaacttgg catagtttgt ccactggaaa atctcaaagc ctttaaccaa
     5281 aggattcctg atttccacag ttctcgtcat cagctctctg gttgctttag ctaatacacc
     5341 ataagcattt tccctactga tgttcatcat ctgagcgtat tggttataag tgaacgatac
     5401 cgtccgttct ttccttgtag ggttttcaat cgtggggttg agtagtgcca cacagcataa
     5461 aattagcttg gtttcatgct ccgttaagtc atagcgacta atcgctagtt catttgcttt
     5521 gaaaacaact aattcagaca tacatctcaa ttggtctagg tgattttaat cactatacca
     5581 attgagatgg gctagtcaat gataattact agtccttttc ccgggagatc tgggtatctg
     5641 taaattctgc tagacctttg ctggaaaact tgtaaattct gctagaccct ctgtaaattc
     5701 cgctagacct ttgtgtgttt tttttgttta tattcaagtg gttataattt atagaataaa
     5761 gaaagaataa aaaaagataa aaagaataga tcccagccct gtgtataact cactacttta
     5821 gtcagttccg cagtattaca aaaggatgtc gcaaacgctg tttgctcctc tacaaaacag
     5881 accttaaaac cctaaaggct taagtagcac cctcgcaagc tcgggcaaat cgctgaatat
     5941 tccttttgtc tccgaccatc aggcacctga gtcgctgtct ttttcgtgac attcagttcg
     6001 ctgcgctcac ggctctggca gtgaatgggg gtaaatggca ctacaggcgc cttttatgga
     6061 ttcatgcaag gaaactaccc ataatacaag aaaagcccgt cacgggcttc tcagggcgtt
     6121 ttatggcggg tctgctatgt ggtgctatct gactttttgc tgttcagcag ttcctgccct
     6181 ctgattttcc agtctgacca cttcggatta tcccgtgaca ggtcattcag actggctaat
     6241 gcacccagta aggcagcggt atcatcaaca ggcttacccg tcttactgtc cctagtgctt
     6301 ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg
     6361 agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctcga tatcaaatta
     6421 cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg
     6481 gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc
     6541 ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac
     6601 gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc
     6661 aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata
     6721 tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc
     6781 agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc
     6841 gtctttcatt gccatacgga attccggatg agcattcatc aggcgggcaa gaatgtgaat
     6901 aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc
     6961 cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc
     7021 tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt
     7081 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat
     7141 ttcattatgg tgaaagttgg aacctcttac gtgccgatca acgtctcatt ttcgccagat
     7201 atc
//
