LOCUS       sequence-350803-        7168 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 7168)
  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 7168)
  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..7168
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      696..1241
                     /direction=RIGHT
                     /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     1142..1161
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1358..1375
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1767..1869
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1870..2529
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1936..1955)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(2750..2797)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2793..2811
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(2849..2893)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(2906..4204)
                     /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"
     protein_bind    4247..4271
                     /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(4272..4290)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(6245..6298)
                     /codon_start=1
                     /product="GroES chaperone mobile loop that interacts with 
                     GroEL"
                     /label=S loop
                     /translation="EVETKSAGGIVLTGSAAA"
     protein_bind    6390..6414
                     /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(6415..6433)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        6479..6556
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(6557..7168,1..471)
                     /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"
ORIGIN
        1 ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc
       61 ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt
      121 gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt
      181 ggtgcggaca tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg
      241 ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg
      301 ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg
      361 aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
      421 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
      481 aattaatgta agttagctca ctcattaggc accgggatct cgaccgatgc ccttgagagc
      541 cttcaaccca gtcagctcct tccggtgggc gcggggcatg actaacatga gaattacaac
      601 ttatatcgta tggggctgac ttcaggtgct acatttgaag agataaattg cactgaaatc
      661 tagaaatatt ttatctgatt aataagatga tcttcttgag atcgttttgg tctgcgcgta
      721 atctcttgct ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga
      781 gctaccaact ctttgaaccg aggtaactgg cttggaggag cgcagtcacc aaaacttgtc
      841 ctttcagttt agccttaacc ggcgcatgac ttcaagacta actcctctaa atcaattacc
      901 agtggctgct gccagtggtg cttttgcatg tctttccggg ttggactcaa gacgatagtt
      961 accggataag gcgcagcggt cggactgaac ggggggttcg tgcatacagt ccagcttgga
     1021 gcgaactgcc tacccggaac tgagtgtcag gcgtggaatg agacaaacgc ggccataaca
     1081 gcggaatgac accggtaaac cgaaaggcag gaacaggaga gcgcacgagg gagccgccag
     1141 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccaccactga tttgagcgtc
     1201 agatttcgtg atgcttgtca ggggggcgga gcctatggaa aaacggcttt gccgcggccc
     1261 tctcacttcc ctgttaagta tcttcctggc atcttccagg aaatctccgc cccgttcgta
     1321 agccatttcc gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc
     1381 ggaatatatc ctgtatcaca tattctgctg acgcaccggt gcagcctttt ttctcctgcc
     1441 acatgaagca cttcactgac accctcatca gtgccaacat agtaagccag tatacactcc
     1501 gctagcgctg atgtccggcg gtgcttttgc cgttacgcac caccccgtca gtagctgaac
     1561 aggagggaca gctgatagaa acagaagcca ctggagcacc tcaaaaacac catcatacac
     1621 taaatcagta agttggcagc atcacccgac gcactttgcg ccgaataaat acctgtgacg
     1681 gaagatcact tcgcagaata aataaatcct ggtgtccctg ttgataccgg gaagccctgg
     1741 gccaactttt ggcgaaaatg agacgttgat cggcacgtaa gaggttccaa ctttcaccat
     1801 aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag
     1861 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
     1921 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
     1981 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
     2041 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggagttccg tatggcaatg
     2101 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
     2161 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
     2221 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
     2281 tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat
     2341 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcactatggg caaatattat
     2401 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
     2461 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     2521 ggggcgtaat ttttttaagg cagttattgg tgcccttaaa cgcctggttg ctacgcctga
     2581 ataagtgata ataagcggat gaatggcaga aattcgaaag caaattcgac ccggtcgtcg
     2641 gttcagggca gggtcgttaa atagccgctt atgtctattg ctggtttacc ggtttattga
     2701 ctaccggaag cagtgtgacc gtgtgcttct caaatgcctg aggtttcagc aaaaaacccc
     2761 tcaagacccg tttagaggcc ccaaggggtt atgctagtta ttgctcagcg gtggcagcag
     2821 cctaggttaa ttaagctgcg ctagtagacg agtccatgtg ctggcgttca aatttcgcag
     2881 cagcggtttc tttaccagac tcgagttacg cctgctggtt catcagctcg ttgaaagtgg
     2941 tttctttttc agtcactttc gctttcgcca gtacagcttc aacagcctgt tcttccagag
     3001 caacattgcg catgttgtcc atcagttctt tgtttttgct gtagaactcg ataacttctt
     3061 tcggatcttc gtacgcagaa gccatctctt cgatcaggcc tttcacgcgc tcctcgtcag
     3121 ctttcagctc gttggtgcgg ataacttcgc ccagcagcag gccaacaact acgcggcgtt
     3181 tagcctgttc ttcgaacagt tcgcgcggca gttccagagc ttgtttttcg ttgccaccga
     3241 aacgctgtgc agcctggcga cgcagaacgt cgatttcgct gtcgatcagc gcagccggta
     3301 cgtcgatgtc gttagctttt accagacctt cgatcgcctg agacttaacg cggttacgga
     3361 tggcgctctt cagctcgcgc tccatgtttt tacgcacttc agcgcgcaga ccttctacgg
     3421 aaccatcttc aacgccgaaa cgtttgatga attctgcagt cagttccggc agttcacgct
     3481 cttcaacttt cttcaggttg atagcgaatt tcgctgcttt acctttcagg ctttctgcgt
     3541 ggtattcttc cgggaaggtc acgtcgatgg tgaactcttc gccagctttg tggcctttga
     3601 taccgtcttc aaagcccggg atcatacgac cctggcccat cgccagtacg aaatcagacg
     3661 ctttaccgcc ttcgaactct tcgccgtcta cagaaccggt gaagtcgatg gttacgcggt
     3721 cttctgcttc aacagcgccg tctttttctt tccaggtcgc ctgctgttta cgcagagtat
     3781 ccagcatgcc gtcaacgtca gcgtcggtca cttcaacgat cggtttttca acttcgatcg
     3841 cttccagacc ctgcagttca acttccggat aaacttcaaa ctctacagag taagtgaagt
     3901 cttcacccag cttgtattcg cccggaacat aagtcggtgc gccagccgga ttgatttttt
     3961 ctttaatgat ggcgtcaatg aagttacggc tcatcaggtc acccagaacg tcctggcgta
     4021 cagacgcgcc ataacgctga gcaacgatat tcattggcac tttgcctttg cggaagccgt
     4081 caatacgtac ttttttcgca acgttgacca gctcgctttt aacagcggtc tcgatgctgt
     4141 cagcagcgac agtaatcgtt acacggcggc caaggccttg agtggtttca actgaaactt
     4201 gcatatgtat atctccttct tatacttaac taatatacta agatggggaa ttgttatccg
     4261 ctcacaattc ccctatagtg agtcgtatta atttcgatta tgcggccgtg tacaatacga
     4321 ttactttctg ttcgacttaa gcattatgcg gccgcaagct tacatcatgc cgcccatgcc
     4381 acccatgccg cccataccgc cagcagcgcc taagtcagct gcatcgtttt tcggcaggtc
     4441 ggtaaccatg cattcggtgg tgatcatcag gccagccaca gaagctgcgt actgcagagc
     4501 agaacgagtt actttggttg gatccaggat acccatgtcg atcatgttgc cgtattcttc
     4561 ggttgctgcg ttgtaaccgt agttgccgtc gccgccttta acggtgttag caacaacaga
     4621 cggttcttcg ccgcagttca atacgatctg acgcagcgga gcttccattg cacgcagtgc
     4681 aactttgata cccacgttct ggtcttcgtt ctgaccacgc aggtcagcca gtttagacgc
     4741 tacgcggatc agcgcaacac caccaccagc aaccacgcct tcttctaccg cagcacgggt
     4801 cgcgtgcagg gcatcttcaa cgcgtgcttt tttctctttc atttcaactt cggtagcagc
     4861 acccactttg ataactgcaa cgccgcctgc cagtttcgct acgcgttcct gcagtttttc
     4921 acggtcgtag tcagaagttg cttcttcaat ctgctgacgg atctgagcaa cacggccctg
     4981 gattgcagct tcttcaccca cgccatcgat gatagtggtg gtgtctttgt tgatcacaac
     5041 acgtttagcc tgacccaggt cttccagggt tgctttttcc agctccatac cgatctcttc
     5101 agagatcacg gtaccgccag tcagggttgc gatatcctgc agcatagctt tacgacgatc
     5161 gccgaagccc ggtgctttaa ccgcagcgac tttcacgatg ccacgcatgg tgttaacaac
     5221 cagagttgcc agcgcttcgc cttctacatc ttcagcgatg atcagcagcg gtttgcctgc
     5281 tttggcaaca gcttccagaa ccggcagcat ttcgcggatg ttggagattt tcttgtcagc
     5341 cagcaggatg aacgggcttt ccagttctac tgcgccagtt tccggcttgt tgatgaagta
     5401 aggagacagg tagccacggt cgaactgcat accttcaacc acgtccagtt cgtcctgcag
     5461 accggtaccg tcttcaacgg tgataacgcc ttctttaccg actttgtcca tcgcttcagc
     5521 gatcagttta cctacggttt cgtcggagtt agcggagatg gtaccaacct gagcaatcgc
     5581 tttagagtca gagcatggta cggacagcgc tttcagttct tcaactgcag cggtaaccgc
     5641 tttgtcgata ccacgtttca ggtccatcgg gttcatgccc gcagcaacag ctttcagacc
     5701 ttcagtgatg atagcctgag ccagtacggt tgcagtggtg gtaccgtcgc ctgcagcgtc
     5761 gtttgcttta gaggcaactt ctttcaccat ctgcgcaccc atattttcga acttgtcttc
     5821 cagttcgatt tcacgagcaa cggaaacacc atctttggtg atggtcggtg caccgaaaga
     5881 tttatccaga actacgttac ggccttttgg accgagggta actttcactg catctgccag
     5941 tacgtttacg ccgcgcagca ttttcacacg agcgtcgtta ccgaatttta cgtctttagc
     6001 tgccattatc tttattcctt aaattcgtat gttcagtgtc gtgcgcggat tacgcttcaa
     6061 caattgccag aatgtcgctt tcggacatga tcaacacttc ttcattgtcg atcttctcag
     6121 atttcacacc gtagccatcg ttgaaaataa cgatgtcgcc aactttcaca tccagcggct
     6181 tcacttcgcc attttcaagg atacggccat tgccgacagc cagcacttcg ccgcgggtgg
     6241 atttagccgc tgcagagccg gtcagaacga tgccgccagc agatttagtt tcaacttctt
     6301 tacgcttgac gatcacgcga tcatgcaatg gacgaatatt ccccatggta tatctcctta
     6361 ttaaagttaa acaaaattat ttctacaggg gaattgttat ccgctcacaa ttcccctata
     6421 gtgagtcgta ttaatttcct aatgcaggag tcgcataagg gagagcgtcg agatcccgga
     6481 caccatcgaa tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca
     6541 attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt
     6601 ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg
     6661 ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca
     6721 actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc
     6781 gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt
     6841 ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct
     6901 cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc
     6961 tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc
     7021 catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt
     7081 cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg
     7141 tctgcgtctg gctggctggc ataaatat
//
