LOCUS       sequence-160356-        4514 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Co-expresses a hisx6 tagged copy of the Ivy protein and the wild 
            type human lysozyme protein, both in the cytoplasmic compartment..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4514)
  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 4514)
  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: pRSF-Ivy-his-hLYZ
FEATURES             Location/Qualifiers
     source          1..4514
                     /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             464..481
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        554..572
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    573..597
                     /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             1051..1095
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1133..1151)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1147..1194
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1424..2239)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1424..2239)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2147..2166
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2240..2331)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2347..3096)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(2357..2374)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2597..2616)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3292..4374)
                     /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     4336..4355
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4375..4452)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(4498..4514,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atggggcagg atgatttaac cattagcagc cttgcaaagg gcgaaaccac
      121 caaagctgca tttaatcaga tggtacaagg gcataagctg cctgcctggg tgatgaaagg
      181 cggtacttat actcccgcac aaaccgtaac gttgggagat gagacgtatc aggtgatgag
      241 cgcgtgcaaa ccgcatgact gtggctcgca acgtatcgct gtgatgtggt ccgagaaatc
      301 taatcagatg acggggctgt tctcgactat tgatgagaaa acgtcgcaag agaaactcac
      361 ctggctgaat gtgaacgatg cgctttcgat tgatggtaaa acggtgttgt tcgcggcgtt
      421 gaccggcagc ctggaaaacc atccggatgg ctttaatttt aaacatcacc atcatcacca
      481 ctaagcttgc ggccgcataa tgcttaagtc gaacagaaag taatcgtatt gtacacggcc
      541 gcataatcga aattaatacg actcactata ggggaattgt gagcggataa caattcccca
      601 tcttagtata ttagttaagt ataagaagga gatatacata tgaaggtctt tgaaaggtgt
      661 gagttggcca gaactctgaa aagattggga atggatggct acaggggaat cagcctagca
      721 aactggatgt gtttggccaa atgggagagt ggttacaaca cacgagctac aaactacaat
      781 gctggagaca gaagcactga ttatgggata tttcagatca atagccgcta ctggtgtaat
      841 gatggcaaaa ccccaggagc agttaatgcc tgtcatttat cctgcagtgc tttgctgcaa
      901 gataacatcg ctgatgctgt agcttgtgca aagagggttg tccgtgatcc acaaggcatt
      961 agagcatggg tggcatggag aaatcgttgt caaaacagag atgtccgtca gtatgttcaa
     1021 ggttgtggag tgtaataact cgagtctggt aaagaaaccg ctgctgcgaa atttgaacgc
     1081 cagcacatgg actcgtctac tagcgcagct taattaacct aggctgctgc caccgctgag
     1141 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     1201 cctcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaaac
     1261 cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac aagctgacga
     1321 ccgggtctcc gcaagtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     1381 tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga
     1441 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     1501 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     1561 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     1621 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     1681 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     1741 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     1801 atacgcggtc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     1861 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     1921 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     1981 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     2041 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     2101 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     2161 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt
     2221 cccgttgaat atggctcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     2281 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggcatgc
     2341 agcgctcttc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag
     2401 cggtgtcagc tcactcaaaa gcggtaatac ggttatccac agaatcaggg gataaagccg
     2461 gaaagaacat gtgagcaaaa agcaaagcac cggaagaagc caacgccgca ggcgtttttc
     2521 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagcca gaggtggcga
     2581 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2641 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2701 gcgctttctc atagctcacg ctgttggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2761 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2821 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cattggtaac
     2881 tgatttagag gactttgtct tgaagttatg cacctgttaa ggctaaactg aaagaacaga
     2941 ttttggtgag tgcggtcctc caacccactt accttggttc aaagagttgg tagctcagcg
     3001 aaccttgaga aaaccaccgt tggtagcggt ggtttttctt tatttatgag atgatgaatc
     3061 aatcggtcta tcaagtcaac gaacagctat tccgttactc tagatttcag tgcaatttat
     3121 ctcttcaaat gtagcacctg aagtcagccc catacgatat aagttgtaat tctcatgtta
     3181 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     3241 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     3301 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3361 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     3421 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     3481 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     3541 gtcttcggta tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca gcccggactc
     3601 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     3661 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     3721 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     3781 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     3841 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     3901 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     3961 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     4021 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     4081 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     4141 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     4201 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     4261 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     4321 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     4381 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     4441 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaaattaa
     4501 tacgactcac tata
//
