LOCUS       sequence-202893-        7450 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Biosynthesis of oxylipins by microbial enzymes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7450)
  TITLE     Bacterial expression plasmids of microbial lipoxygenases originated 
            from Myxococcus xanthus
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7450)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pET28a-mxLOX1
FEATURES             Location/Qualifiers
     source          1..7450
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..144
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2315..2332
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2385..2403)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2399..2446
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2483..2938
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2570..2589)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2780..2801
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3030..3845)
                     /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(3030..3845)
                     /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     3753..3772
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3837..3856
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3967..4555
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4456..4475
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4709..4726
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4741..4880
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4806..4825)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4966..4988
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4982..5173)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5982..7064)
                     /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     7026..7045
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7065..7142)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7348..7367
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 agcggcctgg tgccgcgcgg cagccatatg agcgcgagtg tgacccggag aggtggagct
      181 gatgacagac gttgggacgg gcgcgcccgc ggcatgggga cggggatgat gttcgcaggg
      241 ctgcgccgat ggatgggcgc tctcgggggc aaaggacgcg agagcggaag caacgaggtg
      301 ctcgacgcgg aggagctgtc ccggtggtac tcggggctcg cgctcgagga gcgtctggcc
      361 atcagccgcg agctcgcacc gcgcgtccgg gctgtccgtc ccgcgaggga gcccagcacg
      421 ctgccagccg tcgcggtcgg gcggttggtc ttcgagcagg acgggcccca gggccccatt
      481 cccatgcatc acatcaaggt ggagctgtgg gaccgggact tcggcacgcc ggatgacttc
      541 ctgggggagg ggttcacgga ctcggatgga tgtttttcca ttcgctatga cccggccgac
      601 gcgggcgtga atgacctgcc cgacctggag gtgcgcttct tcgagcccca gcacagcttc
      661 cgcccggatg ggcgggtggt ggaggcatgg tgccgcatcg gctctgagaa ggggccggat
      721 gaccaaggtg ggctccacta cgacttcggc acgctgcggc tgccttactg ggagtacgac
      781 cccacgacgc cgttggcccg gttgcttgtc acggaggagg ggactccgcc cacggcctac
      841 gcgccggggc gcgcgctggc gatgctgaag gcggtggcgc ccatcgagct cgtcaagcgc
      901 cgccacctgc tccagggacg gctgggacag gcgccgagcc tggacaggat tcaggcggac
      961 tatccggagg cgatgacggt gcggatggag cgcgagtcgc cgggctccac gcgcaccgac
     1021 gccttcttcg gagagcggct gctcaacggc atgttctcca ccctcatgga tggcgacccg
     1081 gaggctcccg gggacccgga ggcgttccgg ctctacttcc cgtggaacgc ttacgagcag
     1141 gacggcgtcc actgcctgcc ggacgtggac gtgcggctgc ggctggtgga ggggcggctg
     1201 ttgccggtgc gcatcatcct gggcatgcgc gagccgggcg cgacggcgcc gggctctccc
     1261 gtgacgcggc ggagctacac gcccgcggac ggcgaggcct gggaggccgc caagcggatg
     1321 gcgcgggtga gcgcgacgct ggagacggag ctgggcaacc acctggggca gtgccacttc
     1381 aacgtggagc agtacgccat cgccgcgcac cgcaatctgc gccgcagtcc gctgcgctgg
     1441 ctgctcatgc cgcacctgcg ggaggtggtc ctcatcaacc actccgccaa cggcttcctc
     1501 gtgggaccca ccggctacat cacccgcgcc agcgcgctca ccgagcgcag cgtggagacg
     1561 cggctgttgc acctgatggg cagctacgac tggaagggct tcgcgcccgc gccgcccatc
     1621 tgcgagtcgc accgctacgc acgcgcggcg ggcctgttct ggcggctggt aggggagcac
     1681 gtggatgcct tcttcgccga gcacggcgcg gcgttggagg cgcagtggag cgaggtgcgg
     1741 cgcttctcgg acgacctggt ggggcactcg gcgcccgcct tcgtgtgccg ctacctgcgc
     1801 gcgacggtgc cgggaagggc ggcgccgtgg ttcgtgcgct ccgagcgcat ggacctggac
     1861 gcgaaggtcg ccgcgaccca cgccaaggcc gtcagcgcgg tgacccggac ggacgcgccc
     1921 cagcccggag agatggaggc gctcaagcag ttgtgccggt acgtcatcta cttcgcgacg
     1981 ttccgccatg cctgggccaa caacctccaa tgggacgacg cgggcgaggt gctctatgcg
     2041 tgcctgggcc tgcgctgggg caaggcgggc gcgctgtcga cggaggcgga ccatgacgtc
     2101 gcgccgcctc cggaggaggc cacggagatg ctgtggattt cgtggatgct ctcgaagacg
     2161 agctacggct tcctgctggc caacgaggag gcggacgtgc atccccgctt cgtggagtgc
     2221 ctgcgcgccc acgccgccga gttctccgcg ctggggatgg acatccgcac cgtcagttcc
     2281 cgcatcaata tctaagcttg cggccgcact cgagcaccac caccaccacc actgagatcc
     2341 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     2401 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     2461 tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2521 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2581 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2641 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2701 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2761 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2821 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     2881 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     2941 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3001 caaatatgta tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac
     3061 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     3121 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     3181 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     3241 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc
     3301 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     3361 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
     3421 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     3481 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg
     3541 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     3601 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     3661 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     3721 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     3781 tcagcatcca tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg
     3841 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa
     3901 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3961 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4021 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4081 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     4141 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4201 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4261 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4321 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4381 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4441 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4501 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4561 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4621 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4681 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     4741 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact
     4801 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     4861 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     4921 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     4981 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     5041 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     5101 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     5161 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
     5221 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
     5281 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
     5341 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
     5401 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
     5461 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
     5521 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
     5581 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
     5641 gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
     5701 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
     5761 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
     5821 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     5881 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
     5941 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     6001 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6061 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
     6121 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
     6181 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta
     6241 tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg
     6301 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
     6361 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
     6421 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
     6481 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
     6541 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
     6601 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
     6661 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
     6721 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
     6781 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
     6841 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
     6901 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
     6961 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
     7021 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
     7081 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg
     7141 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag
     7201 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa
     7261 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
     7321 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     7381 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat
     7441 cccgcgaaat
//
