LOCUS       sequence-160364-        4708 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Co-expresses cytoplasmic copies of the Skp protein folding chaperone
            and wild type human lysozyme...
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4708)
  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 4708)
  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-hLYZ
FEATURES             Location/Qualifiers
     source          1..4708
                     /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             1066..1110
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1148..1166)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1162..1209
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1429..2088)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2003..2022
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2089..2191)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2583..2600)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2717..3261)
                     /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(2797..2816)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3486..4568)
                     /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     4530..4549
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4569..4646)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(4692..4708,1..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 acatatgaag
      661 gtctttgaaa ggtgtgagtt ggccagaact ctgaaaagat tgggaatgga tggctacagg
      721 ggaatcagcc tagcaaactg gatgtgtttg gccaaatggg agagtggtta caacacacga
      781 gctacaaact acaatgctgg agacagaagc actgattatg ggatatttca gatcaatagc
      841 cgctactggt gtaatgatgg caaaacccca ggagcagtta atgcctgtca tttatcctgc
      901 agtgctttgc tgcaagataa catcgctgat gctgtagctt gtgcaaagag ggttgtccgt
      961 gatccacaag gcattagagc atgggtggca tggagaaatc gttgtcaaaa cagagatgtc
     1021 cgtcagtatg ttcaaggttg tggagtgtaa taactcgagt ctggtaaaga aaccgctgct
     1081 gcgaaatttg aacgccagca catggactcg tctactagcg cagcttaatt aacctaggct
     1141 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     1201 ggttttttgc tgaaacctca ggcatttgag aagcacacgg tcacactgct tccggtagtc
     1261 aataaaccgg taaaccagca atagacataa gcggctattt aacgaccctg ccctgaaccg
     1321 acgaccgggt cgaatttgct ttcgaatttc tgccattcat ccgcttatta tcacttattc
     1381 aggcgtagca ccaggcgttt aagggcacca ataactgcct taaaaaaatt acgccccgcc
     1441 ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc
     1501 acagacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata
     1561 atatttgccc atagtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc
     1621 aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc
     1681 tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag
     1741 aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc
     1801 atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat
     1861 tgccatacgg aactccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg
     1921 ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac
     1981 ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg
     2041 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt
     2101 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg
     2161 gtgaaagttg gaacctctta cgtgccgatc aacgtctcat tttcgccaaa agttggccca
     2221 gggcttcccg gtatcaacag ggacaccagg atttatttat tctgcgaagt gatcttccgt
     2281 cacaggtatt tattcggcgc aaagtgcgtc gggtgatgct gccaacttac tgatttagtg
     2341 tatgatggtg tttttgaggt gctccagtgg cttctgtttc tatcagctgt ccctcctgtt
     2401 cagctactga cggggtggtg cgtaacggca aaagcaccgc cggacatcag cgctagcgga
     2461 gtgtatactg gcttactatg ttggcactga tgagggtgtc agtgaagtgc ttcatgtggc
     2521 aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt gatacaggat atattccgct
     2581 tcctcgctca ctgactcgct acgctcggtc gttcgactgc ggcgagcgga aatggcttac
     2641 gaacggggcg gagatttcct ggaagatgcc aggaagatac ttaacaggga agtgagaggg
     2701 ccgcggcaaa gccgtttttc cataggctcc gcccccctga caagcatcac gaaatctgac
     2761 gctcaaatca gtggtggcga aacccgacag gactataaag ataccaggcg tttcccctgg
     2821 cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca ttccgctgtt
     2881 atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca gttcgctcca
     2941 agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta tccggtaact
     3001 atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca gccactggta
     3061 attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca
     3121 agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt ggtagctcag
     3181 agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca agagattacg
     3241 cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata tttctagatt
     3301 tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg
     3361 taattctcat gttagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     3421 tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt
     3481 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     3541 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     3601 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     3661 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
     3721 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg
     3781 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     3841 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     3901 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
     3961 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
     4021 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
     4081 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
     4141 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     4201 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg
     4261 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
     4321 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
     4381 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     4441 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     4501 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
     4561 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     4621 aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg
     4681 cattaggaaa ttaatacgac tcactata
//
