LOCUS       sequence-203225-        7037 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 7037)
  TITLE     Bacterial expression plasmids of microbial lipoxygenases originated 
            from Myxococcus xanthus
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7037)
  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            "Unpublished"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: pACYCduet-mxLOX1-EH
FEATURES             Location/Qualifiers
     source          1..7037
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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)."
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        2355..2373
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2374..2398
                     /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             3410..3454
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3492..3510)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3506..3553
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3774..4433)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4348..4367
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4434..4536)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4928..4945)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5062..5607)
                     /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(5142..5161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5832..6914)
                     /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     6876..6895
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6915..6992)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taataaggag atataccatg ggcagcagcc atcaccatca tcaccacagc
      121 caggatccga attcgatgag cgcgagtgtg acccggagag gtggagctga tgacagacgt
      181 tgggacgggc gcgcccgcgg catggggacg gggatgatgt tcgcagggct gcgccgatgg
      241 atgggcgctc tcgggggcaa aggacgcgag agcggaagca acgaggtgct cgacgcggag
      301 gagctgtccc ggtggtactc ggggctcgcg ctcgaggagc gtctggccat cagccgcgag
      361 ctcgcaccgc gcgtccgggc tgtccgtccc gcgagggagc ccagcacgct gccagccgtc
      421 gcggtcgggc ggttggtctt cgagcaggac gggccccagg gccccattcc catgcatcac
      481 atcaaggtgg agctgtggga ccgggacttc ggcacgccgg atgacttcct gggggagggg
      541 ttcacggact cggatggatg tttttccatt cgctatgacc cggccgacgc gggcgtgaat
      601 gacctgcccg acctggaggt gcgcttcttc gagccccagc acagcttccg cccggatggg
      661 cgggtggtgg aggcatggtg ccgcatcggc tctgagaagg ggccggatga ccatggtggg
      721 ctccactacg acttcggcac gctgcggctg ccttactggg agtacgaccc cacgacgccg
      781 ttggcccggt tgcttgtcac ggaggagggg actccgccca cggcctacgc gccggggcgc
      841 gcgctggcga tgctgaaggc ggtggcgccc atcgagctcg tcaagcgccg ccacctgctc
      901 cagggacggc tgggacaggc gccgagcctg gacaggattc aggcggacta tccggaggcg
      961 atgacggtgc ggatggagcg cgagtcgccg ggctccacgc gcaccgacgc cttcttcgga
     1021 gagcggctgc tcaacggcat gttctccacc ctcatggatg gcgacccgga ggctcccggg
     1081 gacccggagg cgttccggct ctacttcccg tggaacgctt acgagcagga cggcgtccac
     1141 tgcctgccgg acgtggacgt gcggctgcgg ctggtggagg ggcggctgtt gccggtgcgc
     1201 atcatcctgg gcatgcgcga gccgggcgcg acggcgccgg gctctcccgt gacgcggcgg
     1261 agctacacgc ccgcggacgg cgaggcctgg gaggccgcca agcggatggc gcgggtgagc
     1321 gcgacgctgg agacggagct gggcaaccac ctggggcagt gccacttcaa cgtggagcag
     1381 tacgccatcg ccgcgcaccg caatctgcgc cgcagtccgc tgcgctggct gctcatgccg
     1441 cacctgcggg aggtggtcct catcaaccac tccgccaacg gcttcctcgt gggacccacc
     1501 ggctacatca cccgcgccag cgcgctcacc gagcgcagcg tggagacgcg gctgttgcac
     1561 ctgatgggca gctacgactg gaagggcttc gcgcccgcgc cgcccatctg cgagtcgcac
     1621 cgctacgcac gcgcggcggg cctgttctgg cggctggtag gggagcacgt ggatgccttc
     1681 ttcgccgagc acggcgcggc gttggaggcg cagtggagcg aggtgcggcg cttctcggac
     1741 gacctggtgg ggcactcggc gcccgccttc gtgtgccgct acctgcgcgc gacggtgccg
     1801 ggaagggcgg cgccgtggtt cgtgcgctcc gagcgcatgg acctggacgc gaaggtcgcc
     1861 gcgacccacg ccaaggccgt cagcgcggtg acccggacgg acgcgcccca gcccggagag
     1921 atggaggcgc tcaagcagtt gtgccggtac gtcatctact tcgcgacgtt ccgccatgcc
     1981 tgggccaaca acctccaatg ggacgacgcg ggcgaggtgc tctatgcgtg cctgggcctg
     2041 cgctggggca aggcgggcgc gctgtcgacg gaggcggacc atgacgtcgc gccgcctccg
     2101 gaggaggcca cggagatgct gtggatttcg tggatgctct cgaagacgag ctacggcttc
     2161 ctgctggcca acgaggaggc ggacgtgcat ccccgcttcg tggagtgcct gcgcgcccac
     2221 gccgccgagt tctccgcgct ggggatggac atccgcaccg tcagttcccg catcaatatc
     2281 taaaagcttg cggccgcata atgcttaagt cgaacagaaa gtaatcgtat tgtacacggc
     2341 cgcataatcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
     2401 atcttagtat attagttaag tataagaagg agatatacat atggctgaca tcacgcatcg
     2461 aaccgtaaag acgaatggca tcaacctgca cctcgcggag gcgggttcag ggccgctcgt
     2521 gttgctcctg catggttggc cggagtcctg gtactcgtgg cgccatcagc ttccggcgtt
     2581 ggcggcggcg gggtaccacg cggtggcccc agacgtccgt ggctacggcc agagcgacaa
     2641 gccggaggcc atcgaggcgt acagcatgaa gcagttggtg ggtgatgcgg ttggcctgct
     2701 ggatgcgctg ggggagagga cggccatcgt catcggccac gactggggct cggcgattgc
     2761 ctggaactgt gccgccctgc atcccgaccg cttccgcgcg gtggtgggaa tgagcgttcc
     2821 ccatctgggg cgcgcgccca tgccgcccat gcagttgttt cagcgcatgt tcggtgagaa
     2881 gtggttctac atcctctact tccaggagcc cggcgtggcc gaggccgagt tcgaggcgga
     2941 cgtcccgagg acggtccggg cgattctcac gggcaccccc ggcttcgatg tgacgaatcc
     3001 cgccgtgctg gcgaagaaga agggggaagg cttcctcgcg aggctcgacg tacctgagac
     3061 gttgcccggc tggctcaccg aggcggacgt cgcgtacttc gcgaaggagc tcgccggcag
     3121 tggattccgg ggcgggctca atcgctaccg caacatggac cgggactggc acgaactgcc
     3181 ggagctggcg acggcggtca tctcgcagcc ggccctctac atcgtcggag agaaggaccc
     3241 ggtgcgtgcc ttctctcccg ttgacccgat gaaggccctg gtgccgaacc tggccgacat
     3301 ccacgtcatt cccggcgcgg gtcattgggt tcagcaggag cacgccgccg aggtgaatgc
     3361 cgcgttgctg gccttcctga agaagctgcc ggcctgactc gagtctggta aagaaaccgc
     3421 tgctgcgaaa tttgaacgcc agcacatgga ctcgtctact agcgcagctt aattaaccta
     3481 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     3541 gaggggtttt ttgctgaaac ctcaggcatt tgagaagcac acggtcacac tgcttccggt
     3601 agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac cctgccctga
     3661 accgacgacc gggtcgaatt tgctttcgaa tttctgccat tcatccgctt attatcactt
     3721 attcaggcgt agcaaccagg cgtttaaggg caccaataac tgccttaaaa aaattacgcc
     3781 ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag
     3841 ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc
     3901 gtataatatt tgcccatagt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt
     3961 aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat attctcaata
     4021 aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg
     4081 tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt
     4141 tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct
     4201 ttcattgcca tacggaactc cggatgagca ttcatcaggc gggcaagaat gtgaataaag
     4261 gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc
     4321 tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta
     4381 cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc cattttagct
     4441 tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca
     4501 ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt ctcattttcg ccaaaagttg
     4561 gcccagggct tcccggtatc aacagggaca ccaggattta tttattctgc gaagtgatct
     4621 tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg atgctgccaa cttactgatt
     4681 tagtgtatga tggtgttttt gaggtgctcc agtggcttct gtttctatca gctgtccctc
     4741 ctgttcagct actgacgggg tggtgcgtaa cggcaaaagc accgccggac atcagcgcta
     4801 gcggagtgta tactggctta ctatgttggc actgatgagg gtgtcagtga agtgcttcat
     4861 gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac aggatatatt
     4921 ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga gcggaaatgg
     4981 cttacgaacg gggcggagat ttcctggaag atgccaggaa gatacttaac agggaagtga
     5041 gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat
     5101 ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5161 ccctggcggc tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc
     5221 gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc
     5281 gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg
     5341 gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca
     5401 ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa
     5461 aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta
     5521 gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag
     5581 attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat aaaatatttc
     5641 tagatttcag tgcaatttat ctcttcaaat gtagcacctg aagtcagccc catacgatat
     5701 aagttgtaat tctcatgtta gtcatgcccc gcgcccaccg gaaggagctg actgggttga
     5761 aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa
     5821 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5881 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt
     5941 ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc
     6001 agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac
     6061 ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatgtccgca
     6121 ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg
     6181 gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa
     6241 ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg
     6301 agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct
     6361 aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg
     6421 tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac
     6481 gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag
     6541 ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct
     6601 tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga
     6661 gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg
     6721 ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc
     6781 agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg
     6841 ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac
     6901 gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata
     6961 ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga
     7021 ctcctgcatt aggaaat
//
