LOCUS       sequence-160362-        6293 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Co-expresses cytoplasmic copies of the Skp and Gro EL/ES protein 
            folding chaperones..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6293)
  AUTHORS   Lamppa JW, Tanyos SA, Griswold KE
  TITLE     Engineering Escherichia coli for soluble expression and single step 
            purification of active human lysozyme.
  JOURNAL   J Biotechnol. 2013 Mar 10;164(1):1-8. doi: 
            10.1016/j.jbiotec.2012.11.007. Epub 2012 Dec 7.
  PUBMED    23220215
REFERENCE   2  (bases 1 to 6293)
  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"; doi: 
            "10.1016/j.jbiotec.2012.11.007"; journalName: "J Biotechnol"; date: 
            "2013-03-10- 10"; volume: "164"; issue: "1"; pages: "1-8"
COMMENT     Sequence Label: pACYC-Skp-GroEL/ES
FEATURES             Location/Qualifiers
     source          1..6293
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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        569..587
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    588..612
                     /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             700..753
                     /codon_start=1
                     /product="GroES chaperone mobile loop that interacts with 
                     GroEL"
                     /label=S loop
                     /translation="EVETKSAGGIVLTGSAAA"
     CDS             2651..2695
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2733..2751)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2747..2794
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3014..3673)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3588..3607
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3674..3776)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4168..4185)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4302..4846)
                     /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(4382..4401)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5071..6153)
                     /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"
     promoter        complement(6154..6231)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(6277..6293,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atggctgaca aaattgcaat cgtcaacatg ggcagcctgt tccagcaggt
      121 agcgcagaaa accggtgttt ctaacacgct ggaaaatgag ttcaaaggcc gtgccagcga
      181 actgcagcgt atggaaaccg atctgcaggc taaaatgaaa aagctgcagt ccatgaaagc
      241 gggcagcgat cgcactaagc tggaaaaaga cgtgatggct cagcgccaga cttttgctca
      301 gaaagcgcag gcttttgagc aggatcgcgc acgtcgttcc aacgaagaac gcggcaaact
      361 ggttactcgt atccagactg ctgtgaaatc cgttgccaac agccaggata tcgatctggt
      421 tgttgatgca aacgccgttg cttacaacag cagcgatgta aaagacatca ctgccgacgt
      481 actgaaacag gttaaataag cttgcggccg cataatgctt aagtcgaaca gaaagtaatc
      541 gtattgtaca cggccgcata atcgaaatta atacgactca ctatagggga attgtgagcg
      601 gataacaatt ccccatctta gtatattagt taagtataag aaggagatat acatatgaat
      661 attcgtccat tgcatgatcg cgtgatcgtc aagcgtaaag aagttgaaac taaatctgct
      721 ggcggcatcg ttctgaccgg ctctgcagcg gctaaatcca cccgcggcga agtgctggct
      781 gtcggcaatg gccgtatcct tgaaaatggc gaagtgaagc cgctggatgt gaaagttggc
      841 gacatcgtta ttttcaacga tggctacggt gtgaaatctg agaagatcga caatgaagaa
      901 gtgttgatca tgtccgaaag cgacattctg gcaattgttg aagcgtaatc cgcgcacgac
      961 actgaacata cgaatttaag gaataaagat aatggcagct aaagacgtaa aattcggtaa
     1021 cgacgctcgt gtgaaaatgc tgcgcggcgt aaacgtactg gcagatgcag tgaaagttac
     1081 cctcggtcca aaaggccgta acgtagttct ggataaatct ttcggtgcac cgaccatcac
     1141 caaagatggt gtttccgttg ctcgtgaaat cgaactggaa gacaagttcg aaaatatggg
     1201 tgcgcagatg gtgaaagaag ttgcctctaa agcaaacgac gctgcaggcg acggtaccac
     1261 cactgcaacc gtactggctc aggctatcat cactgaaggt ctgaaagctg ttgctgcggg
     1321 catgaacccg atggacctga aacgtggtat cgacaaagcg gttaccgctg cagttgaaga
     1381 actgaaagcg ctgtccgtac catgctctga ctctaaagcg attgctcagg ttggtaccat
     1441 ctccgctaac tccgacgaaa ccgtaggtaa actgatcgct gaagcgatgg acaaagtcgg
     1501 taaagaaggc gttatcaccg ttgaagacgg taccggtctg caggacgaac tggacgtggt
     1561 tgaaggtatg cagttcgacc gtggctacct gtctccttac ttcatcaaca agccggaaac
     1621 tggcgcagta gaactggaaa gcccgttcat cctgctggct gacaagaaaa tctccaacat
     1681 ccgcgaaatg ctgccggttc tggaagctgt tgccaaagca ggcaaaccgc tgctgatcat
     1741 cgctgaagat gtagaaggcg aagcgctggc aactctggtt gttaacacca tgcgtggcat
     1801 cgtgaaagtc gctgcggtta aagcaccggg cttcggcgat cgtcgtaaag ctatgctgca
     1861 ggatatcgca accctgactg gcggtaccgt gatctctgaa gagatcggta tggagctgga
     1921 aaaagcaacc ctggaagacc tgggtcaggc taaacgtgtt gtgatcaaca aagacaccac
     1981 cactatcatc gatggcgtgg gtgaagaagc tgcaatccag ggccgtgttg ctcagatccg
     2041 tcagcagatt gaagaagcaa cttctgacta cgaccgtgaa aaactgcagg aacgcgtagc
     2101 gaaactggca ggcggcgttg cagttatcaa agtgggtgct gctaccgaag ttgaaatgaa
     2161 agagaaaaaa gcacgcgttg aagatgccct gcacgcgacc cgtgctgcgg tagaagaagg
     2221 cgtggttgct ggtggtggtg ttgcgctgat ccgcgtagcg tctaaactgg ctgacctgcg
     2281 tggtcagaac gaagaccaga acgtgggtat caaagttgca ctgcgtgcaa tggaagctcc
     2341 gctgcgtcag atcgtattga actgcggcga agaaccgtct gttgttgcta acaccgttaa
     2401 aggcggcgac ggcaactacg gttacaacgc agcaaccgaa gaatacggca acatgatcga
     2461 catgggtatc ctggatccaa ccaaagtaac tcgttctgct ctgcagtacg cagcttctgt
     2521 ggctggcctg atgatcacca ccgaatgcat ggttaccgac ctgccgaaaa acgatgcagc
     2581 tgacttaggc gctgctggcg gtatgggcgg catgggtggc atgggcggca tgatgtaact
     2641 cgagtctggt aaagaaaccg ctgctgcgaa atttgaacgc cagcacatgg actcgtctac
     2701 tagcgcagct taattaacct aggctgctgc caccgctgag caataactag cataacccct
     2761 tggggcctct aaacgggtct tgaggggttt tttgctgaaa cctcaggcat ttgagaagca
     2821 cacggtcaca ctgcttccgg tagtcaataa accggtaaac cagcaataga cataagcggc
     2881 tatttaacga ccctgccctg aaccgacgac cgggtcgaat ttgctttcga atttctgcca
     2941 ttcatccgct tattatcact tattcaggcg tagcaccagg cgtttaaggg caccaataac
     3001 tgccttaaaa aaattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa
     3061 gcattctgcc gacatggaag ccatcacaga cggcatgatg aacctgaatc gccagcggca
     3121 tcagcacctt gtcgccttgc gtataatatt tgcccatagt gaaaacgggg gcgaagaagt
     3181 tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga
     3241 cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg
     3301 ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga
     3361 gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc
     3421 atatcaccag ctcaccgtct ttcattgcca tacggaactc cggatgagca ttcatcaggc
     3481 gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta
     3541 aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa
     3601 atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga
     3661 tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc
     3721 ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt
     3781 ctcattttcg ccaaaagttg gcccagggct tcccggtatc aacagggaca ccaggattta
     3841 tttattctgc gaagtgatct tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg
     3901 atgctgccaa cttactgatt tagtgtatga tggtgttttt gaggtgctcc agtggcttct
     3961 gtttctatca gctgtccctc ctgttcagct actgacgggg tggtgcgtaa cggcaaaagc
     4021 accgccggac atcagcgcta gcggagtgta tactggctta ctatgttggc actgatgagg
     4081 gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa
     4141 tatgtgatac aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg
     4201 actgcggcga gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa
     4261 gatacttaac agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc
     4321 cctgacaagc atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta
     4381 taaagatacc aggcgtttcc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc
     4441 ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc
     4501 agttccgggt aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc
     4561 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa
     4621 gcaccactgg cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc
     4681 cggttaaggc taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc
     4741 tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt
     4801 ttcgttttca gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat
     4861 taatcagata aaatatttct agatttcagt gcaatttatc tcttcaaatg tagcacctga
     4921 agtcagcccc atacgatata agttgtaatt ctcatgttag tcatgccccg cgcccaccgg
     4981 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
     5041 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5101 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5161 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     5221 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     5281 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     5341 cactaccgag atgtccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     5401 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     5461 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     5521 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     5581 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     5641 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     5701 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     5761 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     5821 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     5881 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     5941 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     6001 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     6061 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     6121 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     6181 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     6241 gacgctctcc cttatgcgac tcctgcatta ggaaattaat acgactcact ata
//
