LOCUS       sequence-12922-e        4793 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 4793)
  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 4793)
  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: pBB528
FEATURES             Location/Qualifiers
     source          1..4793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    20..36
                     /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)."
     promoter        complement(join(44..50,51..68,69..74))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    89..110
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(636..1586)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      1634..1856
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     promoter        2201..2303
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2304..2963
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2370..2389)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     3392..3414
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3406..3423
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3407..3423
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3490..3567
                     /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."
     CDS             3568..4650
                     /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     complement(3587..3606)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    4715..4731
                     /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    4748..4764
                     /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)."
     primer_bind     complement(join(4789..4793,1..12))
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(join(4789..4793,1..12))
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
       61 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
      121 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
      181 aacgcgcggg gagaggcggt ttgcgtattg ggcgctattt cttccagaat tgccatgatt
      241 ttttccccac gggaggcgtc actggctccc gtgttgtcgg cagctttgat tcgataagca
      301 gcatcgcctg tttcaggctg tctatgtgtg actgttgagc tgtaacaagt tgtctcaggt
      361 gttcaatttc atgttctagt tgctttgttt tactggtttc acctgttcta ttaggtgtta
      421 catgctgttc atctgttaca ttgtcgatct gttcatggtg aacagcttta aatgcaccaa
      481 aaactcgtaa aagctctgat gtatctatct tttttacacc gttttcatct gtgcatatgg
      541 acagttttcc ctttgatatc taacggtgaa cagttgttct acttttgttt gttagtcttg
      601 atgcttcact gatagataca agagccataa gaacctcaga tccttccgta tttagccagt
      661 atgttctcta gtgtggttcg ttgtttttgc gtgagccatg agaacgaacc attgagatca
      721 tgcttacttt gcatgtcact caaaaatttt gcctcaaaac tggtgagctg aatttttgca
      781 gttaaagcat cgtgtagtgt ttttcttagt ccgttacgta ggtaggaatc tgatgtaatg
      841 gttgttggta ttttgtcacc attcattttt atctggttgt tctcaagttc ggttacgaga
      901 tccatttgtc tatctagttc aacttggaaa atcaacgtat cagtcgggcg gcctcgctta
      961 tcaaccacca atttcatatt gctgtaagtg tttaaatctt tacttattgg tttcaaaacc
     1021 cattggttaa gccttttaaa ctcatggtag ttattttcaa gcattaacat gaacttaaat
     1081 tcatcaaggc taatctctat atttgccttg tgagttttct tttgtgttag ttcttttaat
     1141 aaccactcat aaatcctcat agagtatttg ttttcaaaag acttaacatg ttccagatta
     1201 tattttatga atttttttaa ctggaaaaga taaggcaata tctcttcact aaaaactaat
     1261 tctaattttt cgcttgagaa cttggcatag tttgtccact ggaaaatctc aaagccttta
     1321 accaaaggat tcctgatttc cacagttctc gtcatcagct ctctggttgc tttagctaat
     1381 acaccataag cattttccct actgatgttc atcatctgag cgtattggtt ataagtgaac
     1441 gataccgtcc gttctttcct tgtagggttt tcaatcgtgg ggttgagtag tgccacacag
     1501 cataaaatta gcttggtttc atgctccgtt aagtcatagc gactaatcgc tagttcattt
     1561 gctttgaaaa caactaattc agacatacat ctcaattggt ctaggtgatt ttaatcacta
     1621 taccaattga gatgggctag tcaatgataa ttactagtcc ttttcctttg agttgtgggt
     1681 atctgtaaat tctgctagac ctttgctgga aaacttgtaa attctgctag accctctgta
     1741 aattccgcta gacctttgtg tgtttttttt gtttatattc aagtggttat aatttataga
     1801 ataaagaaag aataaaaaaa gataaaaaga atagatccca gccctgtgta taactcacta
     1861 ctttagtcag ttccgcagta ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa
     1921 aacagacctt aaaaccctaa aggcttaagt agcaccctcg caagctcggg caaatcgctg
     1981 aatattcctt ttgtctccga ccatcaggca cctgagtcgc tgtctttttc gtgacattca
     2041 gttcgctgcg ctcacggctc tggcagtgaa tgggggtaaa tggcactaca ggcgccgaat
     2101 aaatacctgt gacggaagat cacttcgcag aataaataaa tcctggtgtc cctgttgata
     2161 ccgggaagcc ctgggccaac ttttggcgaa aatgagacgt tgatcggcac gtaagaggtt
     2221 ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag ttatcgagat
     2281 tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata
     2341 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
     2401 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
     2461 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
     2521 ttcgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
     2581 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
     2641 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct
     2701 atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt
     2761 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
     2821 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
     2881 atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg
     2941 atgagtggca gggcggggcg taattttttt aaggcagtta ttggtgccct taaacgcctg
     3001 gttgctacgc ctgaataagt gataataagc ggatgaatgg cagaaattcg aaagcaaatt
     3061 cgacccggtc gtcggttcag ggcagggtcg ttaaatagcc gcttatgtct attgctggtt
     3121 taccggttta ttgactaccg gaagcagtgt gaccgtgtgc ttctcaaatg cctgaggcca
     3181 gtttgctcag gctctccccg tggaggtaat aattgacgat atgatcattt attctgcctc
     3241 ccagagcctg ataaaaacgg ttagcgccat tcgccattca ggctgcgcaa ctgttgggaa
     3301 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     3361 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     3421 agtgccaagc ttggctgcag gtcgactcga caattcgcgc gcgaaggcga agcggcatgc
     3481 atttacgttg acaccatcga atggtgcaaa acctttcgcg gtatggcatg atagcgcccg
     3541 gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag
     3601 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
     3661 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc
     3721 gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg
     3781 gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt
     3841 gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg
     3901 cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag
     3961 gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct
     4021 gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg
     4081 gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct
     4141 gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag
     4201 ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa
     4261 atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg
     4321 ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg
     4381 ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag
     4441 gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag
     4501 gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg
     4561 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     4621 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa gttagcgcga
     4681 attgtcgaga aaatttatca aaaagagtgt tgacttgtga gcggataaca atgatactta
     4741 gattcaattg tgagcggata acaatttcac acagaattcg taatcatggt cat
//
