LOCUS       sequence-160361-        5608 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Co-expresses cytoplasmic copies of the Skp and Trigger Factor 
            protein folding chaperones..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5608)
  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 5608)
  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-TF
FEATURES             Location/Qualifiers
     source          1..5608
                     /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             655..1953
                     /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             1966..2010
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2048..2066)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2062..2109
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2329..2988)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2903..2922
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2989..3091)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3483..3500)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3617..4161)
                     /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(3697..3716)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4386..5468)
                     /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     5430..5449
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5469..5546)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(5592..5608,1..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 acatatgcaa
      661 gtttcagttg aaaccactca aggccttggc cgccgtgtaa cgattactgt cgctgctgac
      721 agcatcgaga ccgctgttaa aagcgagctg gtcaacgttg cgaaaaaagt acgtattgac
      781 ggcttccgca aaggcaaagt gccaatgaat atcgttgctc agcgttatgg cgcgtctgta
      841 cgccaggacg ttctgggtga cctgatgagc cgtaacttca ttgacgccat cattaaagaa
      901 aaaatcaatc cggctggcgc accgacttat gttccgggcg aatacaagct gggtgaagac
      961 ttcacttact ctgtagagtt tgaagtttat ccggaagttg aactgcaggg tctggaagcg
     1021 atcgaagttg aaaaaccgat cgttgaagtg accgacgctg acgttgacgg catgctggat
     1081 actctgcgta aacagcaggc gacctggaaa gaaaaagacg gcgctgttga agcagaagac
     1141 cgcgtaacca tcgacttcac cggttctgta gacggcgaag agttcgaagg cggtaaagcg
     1201 tctgatttcg tactggcgat gggccagggt cgtatgatcc cgggctttga agacggtatc
     1261 aaaggccaca aagctggcga agagttcacc atcgacgtga ccttcccgga agaataccac
     1321 gcagaaagcc tgaaaggtaa agcagcgaaa ttcgctatca acctgaagaa agttgaagag
     1381 cgtgaactgc cggaactgac tgcagaattc atcaaacgtt tcggcgttga agatggttcc
     1441 gtagaaggtc tgcgcgctga agtgcgtaaa aacatggagc gcgagctgaa gagcgccatc
     1501 cgtaaccgcg ttaagtctca ggcgatcgaa ggtctggtaa aagctaacga catcgacgta
     1561 ccggctgcgc tgatcgacag cgaaatcgac gttctgcgtc gccaggctgc acagcgtttc
     1621 ggtggcaacg aaaaacaagc tctggaactg ccgcgcgaac tgttcgaaga acaggctaaa
     1681 cgccgcgtag ttgttggcct gctgctgggc gaagttatcc gcaccaacga gctgaaagct
     1741 gacgaggagc gcgtgaaagg cctgatcgaa gagatggctt ctgcgtacga agatccgaaa
     1801 gaagttatcg agttctacag caaaaacaaa gaactgatgg acaacatgcg caatgttgct
     1861 ctggaagaac aggctgttga agctgtactg gcgaaagcga aagtgactga aaaagaaacc
     1921 actttcaacg agctgatgaa ccagcaggcg taactcgagt ctggtaaaga aaccgctgct
     1981 gcgaaatttg aacgccagca catggactcg tctactagcg cagcttaatt aacctaggct
     2041 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     2101 ggttttttgc tgaaacctca ggcatttgag aagcacacgg tcacactgct tccggtagtc
     2161 aataaaccgg taaaccagca atagacataa gcggctattt aacgaccctg ccctgaaccg
     2221 acgaccgggt cgaatttgct ttcgaatttc tgccattcat ccgcttatta tcacttattc
     2281 aggcgtagca ccaggcgttt aagggcacca ataactgcct taaaaaaatt acgccccgcc
     2341 ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc
     2401 acagacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata
     2461 atatttgccc atagtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc
     2521 aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc
     2581 tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag
     2641 aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc
     2701 atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat
     2761 tgccatacgg aactccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg
     2821 ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac
     2881 ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg
     2941 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt
     3001 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg
     3061 gtgaaagttg gaacctctta cgtgccgatc aacgtctcat tttcgccaaa agttggccca
     3121 gggcttcccg gtatcaacag ggacaccagg atttatttat tctgcgaagt gatcttccgt
     3181 cacaggtatt tattcggcgc aaagtgcgtc gggtgatgct gccaacttac tgatttagtg
     3241 tatgatggtg tttttgaggt gctccagtgg cttctgtttc tatcagctgt ccctcctgtt
     3301 cagctactga cggggtggtg cgtaacggca aaagcaccgc cggacatcag cgctagcgga
     3361 gtgtatactg gcttactatg ttggcactga tgagggtgtc agtgaagtgc ttcatgtggc
     3421 aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt gatacaggat atattccgct
     3481 tcctcgctca ctgactcgct acgctcggtc gttcgactgc ggcgagcgga aatggcttac
     3541 gaacggggcg gagatttcct ggaagatgcc aggaagatac ttaacaggga agtgagaggg
     3601 ccgcggcaaa gccgtttttc cataggctcc gcccccctga caagcatcac gaaatctgac
     3661 gctcaaatca gtggtggcga aacccgacag gactataaag ataccaggcg tttcccctgg
     3721 cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca ttccgctgtt
     3781 atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca gttcgctcca
     3841 agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta tccggtaact
     3901 atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca gccactggta
     3961 attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca
     4021 agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt ggtagctcag
     4081 agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca agagattacg
     4141 cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata tttctagatt
     4201 tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg
     4261 taattctcat gttagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     4321 tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt
     4381 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4441 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     4501 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     4561 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
     4621 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg
     4681 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     4741 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     4801 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
     4861 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
     4921 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
     4981 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
     5041 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     5101 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg
     5161 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
     5221 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
     5281 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     5341 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     5401 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
     5461 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     5521 aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg
     5581 cattaggaaa ttaatacgac tcactata
//
