LOCUS       sequence-160363-        7166 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Co-expresses cytoplasmic copies of the Gro EL/ES and Trigger Factor 
            protein folding chaperones..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7166)
  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 7166)
  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-GroEL/ES-TF
FEATURES             Location/Qualifiers
     source          1..7166
                     /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)."
     CDS             119..172
                     /codon_start=1
                     /product="GroES chaperone mobile loop that interacts with 
                     GroEL"
                     /label=S loop
                     /translation="EVETKSAGGIVLTGSAAA"
     promoter        2127..2145
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2146..2170
                     /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             2213..3511
                     /codon_start=1
                     /product="ribosome-associated chaperone from E. coli 
                     (Hoffmann et al., 2010)"
                     /label=Trigger Factor
                     /translation="MQVSVETTQGLGRRVTITVAADSIETAVKSELVNVAKKVRIDGFR
                     KGKVPMNIVAQRYGASVRQDVLGDLMSRNFIDAIIKEKINPAGAPTYVPGEYKLGEDFT
                     YSVEFEVYPEVELQGLEAIEVEKPIVEVTDADVDGMLDTLRKQQATWKEKDGAVEAEDR
                     VTIDFTGSVDGEEFEGGKASDFVLAMGQGRMIPGFEDGIKGHKAGEEFTIDVTFPEEYH
                     AESLKGKAAKFAINLKKVEERELPELTAEFIKRFGVEDGSVEGLRAEVRKNMERELKSA
                     IRNRVKSQAIEGLVKANDIDVPAALIDSEIDVLRRQAAQRFGGNEKQALELPRELFEEQ
                     AKRRVVVGLLLGEVIRTNELKADEERVKGLIEEMASAYEDPKEVIEFYSKNKELMDNMR
                     NVALEEQAVEAVLAKAKVTEKETTFNELMNQQA"
     CDS             3524..3568
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3606..3624)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3620..3667
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3887..4546)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4461..4480
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4547..4649)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(5041..5058)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5175..5719)
                     /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(5255..5274)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5944..7026)
                     /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(7027..7104)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(7150..7166,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atggggaata ttcgtccatt gcatgatcgc gtgatcgtca agcgtaaaga
      121 agttgaaact aaatctgctg gcggcatcgt tctgaccggc tctgcagcgg ctaaatccac
      181 ccgcggcgaa gtgctggctg tcggcaatgg ccgtatcctt gaaaatggcg aagtgaagcc
      241 gctggatgtg aaagttggcg acatcgttat tttcaacgat ggctacggtg tgaaatctga
      301 gaagatcgac aatgaagaag tgttgatcat gtccgaaagc gacattctgg caattgttga
      361 agcgtaatcc gcgcacgaca ctgaacatac gaatttaagg aataaagata atggcagcta
      421 aagacgtaaa attcggtaac gacgctcgtg tgaaaatgct gcgcggcgta aacgtactgg
      481 cagatgcagt gaaagttacc ctcggtccaa aaggccgtaa cgtagttctg gataaatctt
      541 tcggtgcacc gaccatcacc aaagatggtg tttccgttgc tcgtgaaatc gaactggaag
      601 acaagttcga aaatatgggt gcgcagatgg tgaaagaagt tgcctctaaa gcaaacgacg
      661 ctgcaggcga cggtaccacc actgcaaccg tactggctca ggctatcatc actgaaggtc
      721 tgaaagctgt tgctgcgggc atgaacccga tggacctgaa acgtggtatc gacaaagcgg
      781 ttaccgctgc agttgaagaa ctgaaagcgc tgtccgtacc atgctctgac tctaaagcga
      841 ttgctcaggt tggtaccatc tccgctaact ccgacgaaac cgtaggtaaa ctgatcgctg
      901 aagcgatgga caaagtcggt aaagaaggcg ttatcaccgt tgaagacggt accggtctgc
      961 aggacgaact ggacgtggtt gaaggtatgc agttcgaccg tggctacctg tctccttact
     1021 tcatcaacaa gccggaaact ggcgcagtag aactggaaag cccgttcatc ctgctggctg
     1081 acaagaaaat ctccaacatc cgcgaaatgc tgccggttct ggaagctgtt gccaaagcag
     1141 gcaaaccgct gctgatcatc gctgaagatg tagaaggcga agcgctggca actctggttg
     1201 ttaacaccat gcgtggcatc gtgaaagtcg ctgcggttaa agcaccgggc ttcggcgatc
     1261 gtcgtaaagc tatgctgcag gatatcgcaa ccctgactgg cggtaccgtg atctctgaag
     1321 agatcggtat ggagctggaa aaagcaaccc tggaagacct gggtcaggct aaacgtgttg
     1381 tgatcaacaa agacaccacc actatcatcg atggcgtggg tgaagaagct gcaatccagg
     1441 gccgtgttgc tcagatccgt cagcagattg aagaagcaac ttctgactac gaccgtgaaa
     1501 aactgcagga acgcgtagcg aaactggcag gcggcgttgc agttatcaaa gtgggtgctg
     1561 ctaccgaagt tgaaatgaaa gagaaaaaag cacgcgttga agatgccctg cacgcgaccc
     1621 gtgctgcggt agaagaaggc gtggttgctg gtggtggtgt tgcgctgatc cgcgtagcgt
     1681 ctaaactggc tgacctgcgt ggtcagaacg aagaccagaa cgtgggtatc aaagttgcac
     1741 tgcgtgcaat ggaagctccg ctgcgtcaga tcgtattgaa ctgcggcgaa gaaccgtctg
     1801 ttgttgctaa caccgttaaa ggcggcgacg gcaactacgg ttacaacgca gcaaccgaag
     1861 aatacggcaa catgatcgac atgggtatcc tggatccaac caaagtaact cgttctgctc
     1921 tgcagtacgc agcttctgtg gctggcctga tgatcaccac cgaatgcatg gttaccgacc
     1981 tgccgaaaaa cgatgcagct gacttaggcg ctgctggcgg tatgggcggc atgggtggca
     2041 tgggcggcat gatgtaagct tgcggccgca taatgcttaa gtcgaacaga aagtaatcgt
     2101 attgtacacg gccgcataat cgaaattaat acgactcact ataggggaat tgtgagcgga
     2161 taacaattcc ccatcttagt atattagtta agtataagaa ggagatatac atatgcaagt
     2221 ttcagttgaa accactcaag gccttggccg ccgtgtaacg attactgtcg ctgctgacag
     2281 catcgagacc gctgttaaaa gcgagctggt caacgttgcg aaaaaagtac gtattgacgg
     2341 cttccgcaaa ggcaaagtgc caatgaatat cgttgctcag cgttatggcg cgtctgtacg
     2401 ccaggacgtt ctgggtgacc tgatgagccg taacttcatt gacgccatca ttaaagaaaa
     2461 aatcaatccg gctggcgcac cgacttatgt tccgggcgaa tacaagctgg gtgaagactt
     2521 cacttactct gtagagtttg aagtttatcc ggaagttgaa ctgcagggtc tggaagcgat
     2581 cgaagttgaa aaaccgatcg ttgaagtgac cgacgctgac gttgacggca tgctggatac
     2641 tctgcgtaaa cagcaggcga cctggaaaga aaaagacggc gctgttgaag cagaagaccg
     2701 cgtaaccatc gacttcaccg gttctgtaga cggcgaagag ttcgaaggcg gtaaagcgtc
     2761 tgatttcgta ctggcgatgg gccagggtcg tatgatcccg ggctttgaag acggtatcaa
     2821 aggccacaaa gctggcgaag agttcaccat cgacgtgacc ttcccggaag aataccacgc
     2881 agaaagcctg aaaggtaaag cagcgaaatt cgctatcaac ctgaagaaag ttgaagagcg
     2941 tgaactgccg gaactgactg cagaattcat caaacgtttc ggcgttgaag atggttccgt
     3001 agaaggtctg cgcgctgaag tgcgtaaaaa catggagcgc gagctgaaga gcgccatccg
     3061 taaccgcgtt aagtctcagg cgatcgaagg tctggtaaaa gctaacgaca tcgacgtacc
     3121 ggctgcgctg atcgacagcg aaatcgacgt tctgcgtcgc caggctgcac agcgtttcgg
     3181 tggcaacgaa aaacaagctc tggaactgcc gcgcgaactg ttcgaagaac aggctaaacg
     3241 ccgcgtagtt gttggcctgc tgctgggcga agttatccgc accaacgagc tgaaagctga
     3301 cgaggagcgc gtgaaaggcc tgatcgaaga gatggcttct gcgtacgaag atccgaaaga
     3361 agttatcgag ttctacagca aaaacaaaga actgatggac aacatgcgca atgttgctct
     3421 ggaagaacag gctgttgaag ctgtactggc gaaagcgaaa gtgactgaaa aagaaaccac
     3481 tttcaacgag ctgatgaacc agcaggcgta actcgagtct ggtaaagaaa ccgctgctgc
     3541 gaaatttgaa cgccagcaca tggactcgtc tactagcgca gcttaattaa cctaggctgc
     3601 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     3661 ttttttgctg aaacctcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa
     3721 taaaccggta aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac
     3781 gaccgggtcg aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag
     3841 gcgtagcacc aggcgtttaa gggcaccaat aactgcctta aaaaaattac gccccgccct
     3901 gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac
     3961 agacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat
     4021 atttgcccat agtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa
     4081 aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt
     4141 tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa
     4201 actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat
     4261 ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg
     4321 ccatacggaa ctccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat
     4381 aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg
     4441 tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc
     4501 attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag
     4561 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
     4621 gaaagttgga acctcttacg tgccgatcaa cgtctcattt tcgccaaaag ttggcccagg
     4681 gcttcccggt atcaacaggg acaccaggat ttatttattc tgcgaagtga tcttccgtca
     4741 caggtattta ttcggcgcaa agtgcgtcgg gtgatgctgc caacttactg atttagtgta
     4801 tgatggtgtt tttgaggtgc tccagtggct tctgtttcta tcagctgtcc ctcctgttca
     4861 gctactgacg gggtggtgcg taacggcaaa agcaccgccg gacatcagcg ctagcggagt
     4921 gtatactggc ttactatgtt ggcactgatg agggtgtcag tgaagtgctt catgtggcag
     4981 gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga tacaggatat attccgcttc
     5041 ctcgctcact gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga
     5101 acggggcgga gatttcctgg aagatgccag gaagatactt aacagggaag tgagagggcc
     5161 gcggcaaagc cgtttttcca taggctccgc ccccctgaca agcatcacga aatctgacgc
     5221 tcaaatcagt ggtggcgaaa cccgacagga ctataaagat accaggcgtt tcccctggcg
     5281 gctccctcgt gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat
     5341 ggccgcgttt gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag
     5401 ctggactgta tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat
     5461 cgtcttgagt ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat
     5521 tgatttagag gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag
     5581 ttttggtgac tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag
     5641 aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg
     5701 cagaccaaaa cgatctcaag aagatcatct tattaatcag ataaaatatt tctagatttc
     5761 agtgcaattt atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgta
     5821 attctcatgt tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     5881 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
     5941 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     6001 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     6061 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     6121 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     6181 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg
     6241 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     6301 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     6361 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     6421 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     6481 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     6541 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     6601 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     6661 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     6721 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     6781 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     6841 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     6901 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     6961 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     7021 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     7081 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca
     7141 ttaggaaatt aatacgactc actata
//
