LOCUS       sequence-207891-        6916 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 6916)
  TITLE     Bacterial expression plasmids of microbial lipoxygenases originated 
            from Myxococcus xanthus
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6916)
  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            "Unpublished"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-mxLOX2-EH
FEATURES             Location/Qualifiers
     source          1..6916
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..25
                     /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             81..98
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        2215..2233
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2234..2258
                     /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             3270..3314
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3352..3370)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3366..3413
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3634..4293)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4208..4227
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4294..4396)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4788..4805)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4922..5467)
                     /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(5002..5021)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5692..6774)
                     /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     6736..6755
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6775..6852)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        6898..6916
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggaattgtga gcggataaca attcccctgt agaaataatt ttgtttaact ttaataagga
       61 gatataccat gggcagcagc catcaccatc atcaccacag ccaggatccg aattcatgac
      121 tgtcgagtac aaactcacga ttcggacggg cacgaagctc ggcgcgggga cggatgccga
      181 catttccatc gtcctcgtgg gcacgcgagg agagagcgcc ccccgcgtgc tggacaagca
      241 cttccacaac gacttcgagg cgggcgcgga ggacgtctac gccctctcct ccgaggacct
      301 cggtgacctg gtcctgttgc gcttcagcaa cgcgggcggc gtggcggccg actggctgct
      361 cgactgggca atcgtcacgg ccggggagaa gcagtggcac ttccctttct accggtgggt
      421 gctgagtggc gccacggtcg acgtgctcga ggggaccgcg aagctcgctc gccaggcaag
      481 cagcgagcgc gagtccacgg cgcggcgcga gctgctcgag gcccgccagc ggatgtaccc
      541 gtggcgcgcg cctgagatga ccgaggggct tcccggcgcg ctcgacctcc gtgaggggag
      601 gccgctgccg aaggacgagc tctaccgggg cctgacggag ggcagctacg aagtggtcat
      661 cgcgaagacg ctggcggcca tcaagctgaa cctgcccatg ctgacccgcg cctggaacgg
      721 gctggtggac atcttcgact tcttcaaaca cctggaggtg ccccagctcg cccagcgctg
      781 gaaggacgac ctcgagttcg cgcggcaggc cgtccagggc atcgcccccc tccacatcac
      841 gctcgtcccc agtctgccgc agggcatgcc gctcaccgac gacgacgtcc ggggcctctt
      901 gtcgcccggc accacgctgg ccagggcgct cggcgccaag cgcatcttcc tgatcgactt
      961 cgaaatcctc gacgacatca ggatgtaccg gaaggtcggc gaggacggag tcgaggagcg
     1021 gcgctgggct cccgcggcgc gctgcctgct gtacctggat gaccagcgtc aactgcgacc
     1081 cctggccatc cagctcgggc gggacgccca gaaggaccct gtcttcacgc cgaacgacga
     1141 cgcgtacgac tggctcgccg cgaaaatcta cctccggtgc agcgagggca actcgcacca
     1201 gatggtgtcg cacgcgctgc gcacacactt cgtggcggag ccgttcgtca tggcgacgat
     1261 gcgcaacctg ccggacccgc accccgtcta caaactgctg cggcggcact tccgctacac
     1321 gctcgccatc aacgagggcg cacgcaaggg cctgctcgac gcaggcgggg tgttcgacga
     1381 cttcatcgcg acaggcggcc ccgacaaggg ccacctccag ttgggcaaga agggcttcca
     1441 gcgctggacg ctggcggaca acaagccccg tgctgacctg gagcggcggg gcgtgctgga
     1501 ccctgccgtg ctccccaact acccgtaccg ggacgacgcc ctgcccttgt gggacgcgtt
     1561 cgaggagtac gtcggcggcg tcctcaggca cttctaccgg accgatgccg acctcgaggc
     1621 cgacaccgag atgcagcaat ggtggaagga cctcaccgag cacgggctgc ccgtggacaa
     1681 gctgccctgc cgggagctgc gccgcgtcga cgacctggtc gacatcctca ccaccgtcct
     1741 cttcacggtc agcgtgcagc acgcggcggt gaactacctg caatacgagc actacgcctt
     1801 cgtaccgaat gcgcccctga gcatgcgccg ggagccaccc cgccagaagg ggacgctgcg
     1861 tgcagaggac atccccgaga tgattcccac caagtcccag atgctctggc aggtcgccat
     1921 cggccgggcg ctctccagct tcggagacga cgaggagtac ctgctgcacg agggcggctg
     1981 gcgcgaggag tacttccacg aaccggagct ggtggccatc cgccagcggt tccaggagcg
     2041 cctgcgcgcc cagcgcgagg cggtggaggc gcgcaacgcg ggcgccgagg tgccctacac
     2101 catcctgcgt cccgaccgga ttccctgcgg catcaccgtc tgaaagcttg cggccgcata
     2161 atgcttaagt cgaacagaaa gtaatcgtat tgtacacggc cgcataatcg aaattaatac
     2221 gactcactat aggggaattg tgagcggata acaattcccc atcttagtat attagttaag
     2281 tataagaagg agatatacat atggctgaca tcacgcatcg aaccgtaaag acgaatggca
     2341 tcaacctgca cctcgcggag gcgggttcag ggccgctcgt gttgctcctg catggttggc
     2401 cggagtcctg gtactcgtgg cgccatcagc ttccggcgtt ggcggcggcg gggtaccacg
     2461 cggtggcccc agacgtccgt ggctacggcc agagcgacaa gccggaggcc atcgaggcgt
     2521 acagcatgaa gcagttggtg ggtgatgcgg ttggcctgct ggatgcgctg ggggagagga
     2581 cggccatcgt catcggccac gactggggct cggcgattgc ctggaactgt gccgccctgc
     2641 atcccgaccg cttccgcgcg gtggtgggaa tgagcgttcc ccatctgggg cgcgcgccca
     2701 tgccgcccat gcagttgttt cagcgcatgt tcggtgagaa gtggttctac atcctctact
     2761 tccaggagcc cggcgtggcc gaggccgagt tcgaggcgga cgtcccgagg acggtccggg
     2821 cgattctcac gggcaccccc ggcttcgatg tgacgaatcc cgccgtgctg gcgaagaaga
     2881 agggggaagg cttcctcgcg aggctcgacg tacctgagac gttgcccggc tggctcaccg
     2941 aggcggacgt cgcgtacttc gcgaaggagc tcgccggcag tggattccgg ggcgggctca
     3001 atcgctaccg caacatggac cgggactggc acgaactgcc ggagctggcg acggcggtca
     3061 tctcgcagcc ggccctctac atcgtcggag agaaggaccc ggtgcgtgcc ttctctcccg
     3121 ttgacccgat gaaggccctg gtgccgaacc tggccgacat ccacgtcatt cccggcgcgg
     3181 gtcattgggt tcagcaggag cacgccgccg aggtgaatgc cgcgttgctg gccttcctga
     3241 agaagctgcc ggcctgactc gagtctggta aagaaaccgc tgctgcgaaa tttgaacgcc
     3301 agcacatgga ctcgtctact agcgcagctt aattaaccta ggctgctgcc accgctgagc
     3361 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaac
     3421 ctcaggcatt tgagaagcac acggtcacac tgcttccggt agtcaataaa ccggtaaacc
     3481 agcaatagac ataagcggct atttaacgac cctgccctga accgacgacc gggtcgaatt
     3541 tgctttcgaa tttctgccat tcatccgctt attatcactt attcaggcgt agcaaccagg
     3601 cgtttaaggg caccaataac tgccttaaaa aaattacgcc ccgccctgcc actcatcgca
     3661 gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg
     3721 aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatagt
     3781 gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact
     3841 cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc
     3901 caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc
     3961 gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta
     4021 acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca tacggaactc
     4081 cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt
     4141 atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt
     4201 acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc
     4261 aacggtggta tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct
     4321 cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc
     4381 tcttacgtgc cgatcaacgt ctcattttcg ccaaaagttg gcccagggct tcccggtatc
     4441 aacagggaca ccaggattta tttattctgc gaagtgatct tccgtcacag gtatttattc
     4501 ggcgcaaagt gcgtcgggtg atgctgccaa cttactgatt tagtgtatga tggtgttttt
     4561 gaggtgctcc agtggcttct gtttctatca gctgtccctc ctgttcagct actgacgggg
     4621 tggtgcgtaa cggcaaaagc accgccggac atcagcgcta gcggagtgta tactggctta
     4681 ctatgttggc actgatgagg gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct
     4741 gcaccggtgc gtcagcagaa tatgtgatac aggatatatt ccgcttcctc gctcactgac
     4801 tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg gggcggagat
     4861 ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg gcaaagccgt
     4921 ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca aatcagtggt
     4981 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc tccctcgtgc
     5041 gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt
     5101 ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct ggactgtatg
     5161 cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5221 aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg atttagagga
     5281 gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg
     5341 cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa
     5401 accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg
     5461 atctcaagaa gatcatctta ttaatcagat aaaatatttc tagatttcag tgcaatttat
     5521 ctcttcaaat gtagcacctg aagtcagccc catacgatat aagttgtaat tctcatgtta
     5581 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     5641 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     5701 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5761 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     5821 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     5881 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     5941 gtcttcggta tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca gcccggactc
     6001 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     6061 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     6121 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     6181 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     6241 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     6301 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     6361 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     6421 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     6481 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     6541 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     6601 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     6661 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     6721 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     6781 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     6841 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaaattaa
     6901 tacgactcac tatagg
//
