LOCUS       sequence-202892-        7332 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 7332)
  TITLE     Bacterial expression plasmids of microbial lipoxygenases originated 
            from Myxococcus xanthus
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7332)
  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            "Unpublished"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-mxLOX2
FEATURES             Location/Qualifiers
     source          1..7332
                     /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             2197..2214
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2267..2285)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2281..2328
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2365..2820
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2452..2471)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2662..2683
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2912..3727)
                     /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(2912..3727)
                     /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     3635..3654
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3719..3738
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3849..4437
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4338..4357
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4591..4608
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4623..4762
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4688..4707)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4848..4870
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4864..5055)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5864..6946)
                     /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     6908..6927
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6947..7024)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7230..7249
                     /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 actgtcgagt acaaactcac gattcggacg
      181 ggcacgaagc tcggcgcggg gacggatgcc gacatttcca tcgtcctcgt gggcacgcga
      241 ggagagagcg ccccccgcgt gctggacaag cacttccaca acgacttcga ggcgggcgcg
      301 gaggacgtct acgccctctc ctccgaggac ctcggtgacc tggtcctgtt gcgcttcagc
      361 aacgcgggcg gcgtggcggc cgactggctg ctcgactggg caatcgtcac ggccggagag
      421 aagcagtggc acttcccttt ctaccggtgg gtgctgagtg gcgccacggt cgacgtgctc
      481 gaggggaccg cgaagctcgc tcgccaggca agcagcgagc gcgagtccac ggcgcggcgc
      541 gagctgctcg aggcccgcca gcggatgtac ccgtggcgcg cgcctgagat gaccgagggg
      601 cttcccggcg cgctcgacct ccgtgagggg aggccgctgc cgaaggacga gctctaccgg
      661 ggcctgacgg agggcagcta cgaagtggtc atcgcgaaga cgctggcggc catcaagctg
      721 aacctgccca tgctgacccg cgcctggaac gggctggtgg acatcttcga cttcttcaaa
      781 cacctggagg tgccccagct cgcccagcgc tggaaggacg acctcgagtt cgcgcggcag
      841 gccgtccagg gcatcgcccc cctccacatc acgctcgtcc ccagtctgcc gcagggcatg
      901 ccgctcaccg acgacgacgt ccggggcctc ttgtcgcccg gcaccacgct ggccagggcg
      961 ctcgacgcca agcgcatctt cctgatcgac ttcgaaatcc tcgacgacat caggatgtac
     1021 cggaaggtcg gcgaggacgg agtcgaggag cggcgctggg ctcccgcggc gcgctgcctg
     1081 ctgtacctgg atgaccagcg tcaactgcga cccctggcca tccagctcgg gcgggacgcc
     1141 cagaaggacc ctgtcttcac gccgaacgac gacgcgtacg actggctcgc cgcgaaaatc
     1201 tacctccggt gcagcgaggg caactcgcac cagatggtgt cgcacgcgct gcgcacacac
     1261 ttcgtggcgg agccgttcgt catggcgacg atgcgcaacc tgccggaccc gcaccccgtc
     1321 tacaaactgc tgcggcggca cttccgctac acgctcgcca tcaacgaggg cgcacgcaag
     1381 ggcctgctcg acgcaggcgg ggtgttcgac gacttcatcg cgacaggcgg ccccgacaag
     1441 ggccacctcc agttgggcaa gaagggcttc cagcgctgga cgctggcgga caacaagccc
     1501 cgtgctgacc tggagcggcg gggcgtgctg gaccctgccg tgctccccaa ctacccgtac
     1561 cgggacgacg ccctgccctt gtgggacgcg ttcgaggagt acgtcggcgg cgtcctcagg
     1621 cacttctacc ggaccgatgc cgacctcgag gccgacaccg agatgcagca atggtggaag
     1681 gacctcaccg agcacgggct gcccgtggac aagctgccct gccgggagct gcgccgcgtc
     1741 gacgacctgg tcgacatcct caccaccgtc ctcttcacgg tcagcgtgca gcacgcggcg
     1801 gtgaactacc tgcaatacga gcactacgcc ttcgtaccga atgcgcccct gagcatgcgc
     1861 cgggagccac cccgccagaa ggggacgctg cgtgcagagg acatccccga gatgattccc
     1921 accaagtccc agatgctctg gcaggtcgcc atcggccggg cgctctccag cttcggagac
     1981 gacgaggagt acctgctgca cgagggcggc tggcgcgagg agtacttcca cgaaccggag
     2041 ctggtggcca tccgccagcg gttccaggag cgcctgcgcg cccagcgcga ggcggtggag
     2101 gcgcgcaacg cgggcgccga ggtgccctac accatcctgc gtcccgaccg gattccctgc
     2161 ggcatcaccg tctgaaagct tgcggccgca ctcgagcacc accaccacca ccactgagat
     2221 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     2281 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
     2341 actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2401 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2461 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2521 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2581 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2641 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2701 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2761 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     2821 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2881 ttcaaatatg tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa
     2941 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
     3001 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
     3061 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
     3121 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat
     3181 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
     3241 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc
     3301 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
     3361 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc
     3421 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
     3481 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
     3541 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     3601 atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     3661 aatcagcatc catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat
     3721 ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgacc
     3781 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3841 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3901 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3961 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     4021 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4081 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4141 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4201 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4261 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4321 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4381 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4441 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4501 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     4561 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     4621 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca
     4681 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
     4741 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     4801 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     4861 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     4921 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     4981 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     5041 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
     5101 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
     5161 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
     5221 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
     5281 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
     5341 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
     5401 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
     5461 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg
     5521 tggggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc
     5581 agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat
     5641 catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac
     5701 ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc
     5761 ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc
     5821 cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca
     5881 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5941 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
     6001 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
     6061 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg
     6121 tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg
     6181 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
     6241 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
     6301 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
     6361 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
     6421 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
     6481 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
     6541 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
     6601 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
     6661 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
     6721 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
     6781 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
     6841 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
     6901 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga
     6961 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg
     7021 tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
     7081 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc
     7141 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
     7201 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     7261 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg
     7321 atcccgcgaa at
//
