LOCUS       sequence-67072-e        6656 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BglBrick plasmid (=pBbB8k-P450) coding for a cytochrome P450 and its
            associated reductases to oxidize limonene to perillyl alcohol in E. 
            coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6656)
  AUTHORS   Alonso-Gutierrez J, Chan R, Batth TS, Adams PD, Keasling JD, Petzold
            CJ, Lee TS
  TITLE     Metabolic engineering of Escherichia coli for limonene and perillyl 
            alcohol production.
  JOURNAL   Metab Eng. 2013 May 29;19C:33-41. doi: 10.1016/j.ymben.2013.05.004.
  PUBMED    23727191
REFERENCE   2  (bases 1 to 6656)
  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: "Metab 
            Eng."; date: "2013-05-29"; volume: "19C:33-41. doi"; pages: " 
            10.1016/j.ymben.2013.05.004"
COMMENT     Sequence Label: pJBEI-6411
FEATURES             Location/Qualifiers
     source          1..6656
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(7..885)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        912..1196
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1119..1138
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     terminator      4171..4242
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4258..4285
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4352..5014)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVLAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     terminator      complement(5601..5695)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(5726..6520)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5837..5856)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6447..6466
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtcttat gacaacttga cggctacatc attcactttt tcttcacaac cggcacggaa
       61 ctcgctcggg ctggccccgg tgcatttttt aaatacccgc gagaaataga gttgatcgtc
      121 aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca aaagcagctt
      181 cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta actgctggcg
      241 gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc
      301 aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta cccgattatc
      361 catcggtgga tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag
      421 cagatttatc gccagcagct ccgaatagcg cccttcccct tgcccggcgt taatgatttg
      481 cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga accccgtatt
      541 ggcaaatatt gacggccagt taagccattc atgccagtag gcgcgcggac gaaagtaaac
      601 ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa tctctcctgg
      661 cgggaacagc aaaatatcac ccggtcggca aacaaattct cgtccctgat ttttcaccac
      721 cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat
      781 aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt
      841 aaacgagtat cccggcagca ggggatcatt ttgcgcttca gccatacttt tcatactccc
      901 gccattcaga gaagaaacca attgtccata ttgcatcaga cattgccgtc actgcgtctt
      961 ttactggctc ttctcgctaa ccaaaccggt aaccccgctt attaaaagca ttctgtaaca
     1021 aagcgggacc aaagccatga caaaaacgcg taacaaaagt gtctataatc acggcagaaa
     1081 agtccacatt gattatttgc acggcgtcac actttgctat gccatagcat ttttatccat
     1141 aagattagcg gattctacct gacgcttttt atcgcaactc tctactgttt ctccataccc
     1201 gtttttttgg gaattcaaaa gatctaggag gataaagaaa tgactgaaat gactgtcgcg
     1261 gcttcggacg caactaatgc cgcctacggc atggccctgg aagatattga tgttagcaat
     1321 ccggttctgt tccgcgataa tacgtggcac ccctatttca aaaggctgcg tgaggaagac
     1381 ccagtgcact attgtaagtc ctctatgttt ggtccatact ggagtgttac caaatatcgt
     1441 gatataatgg ccgtggaaac gaacccgaaa gtattttcaa gcgaggctaa atcaggtggt
     1501 attaccataa tggatgataa cgccgctgct agcctgccta tgtttatcgc gatggacccc
     1561 ccgaaacatg atgttcagcg aaaaaccgta tcccctattg ttgcgccgga gaatttagcc
     1621 acgatggagt cagtcatacg ccagagaacg gctgacttgc ttgatggttt acccataaat
     1681 gaagaatttg actgggttca tcgcgtctcc attgagttaa ccaccaaaat gctggcgacc
     1741 ctgtttgact tcccctggga cgatagagcg aaattgactc gctggtccga tgtgaccaca
     1801 gctttacccg gaggtggtat aattgatagc gaagaacagc gtatggccga gttgatggag
     1861 tgcgctacgt acttcaccga actgtggaat cagcgtgtta acgctgagcc aaaaaatgat
     1921 ctgatctcta tgatggcaca ctcggagagt actagacata tggctccgga ggaatactta
     1981 gggaacatcg tgctgctcat cgtaggaggg aatgatacga cccgtaactc catgacgggt
     2041 ggtgtacttg ccctaaacga atttcccgat gaatatcgga agctaagtgc gaatcctgca
     2101 ttgatctcgt caatggtgtc cgaaatcatt cggtggcaga cccctctctc ccatatgcgc
     2161 cggacggctc ttgaagacat agagtttggt ggaaaacata ttagacaagg agacaaggtg
     2221 gttatgtggt atgtttccgg caatcgagat ccagaagcta tcgacaaccc tgatacattt
     2281 attatcgatc gtgcaaagcc acgtcaacac ctgtcttttg gcttcggtat tcatcgttgc
     2341 gttggtaatc gcctggcgga attacaatta aatattctgt gggaagagat tctgaaacga
     2401 tggccagacc cccttcagat tcaagtttta caggaaccaa cgcgtgttct gagtcctttt
     2461 gtaaaaggtt atgaatcgct gccagtacgc atcaacgcgt aaggatctag gaggataaag
     2521 aaatgataca tacaggcgtc acggaggctg ttgtggtcgt cggtgctggg caggcgggag
     2581 cacagactgt gacaagtttg cgacagcgcg gttttgaagg tcagattacg ctgttaggag
     2641 atgaacccgc gttaccgtat cagcgcccac cgttatccaa agcatttttg gctggcacac
     2701 tgccactgga tcgcctgtat ttacgcccag ccgcgtttta ccaacaggca catgtggatg
     2761 tcatggtcga tacaggtgtc tcggaactag ataccgaaaa tcgtcgtatc cggctgacag
     2821 atggacgagc gatatctttt gaccatctgg ttttagccac cggggggagg cccagacctt
     2881 tagcctgtcc cggagcggac cacccccgcg ttcattattt gcgtacagtc accgatgttg
     2941 atcgtattcg gtcacagttt catcctggca cccgacttgt attagtgggg ggggggtaca
     3001 tagggttaga aattgcagcg gtagccgccg aattaggttt gactgtaaca gtgcttgagg
     3061 cccagacaac tgtgttggcc cgcgttacgt gtccaaccgt cgcacgcttt ttcgagcata
     3121 cacaccggcg cgcgggtgtc accattcgct gcgccacgac cgtgacacgg attcatgatt
     3181 cttcgagtac tgctcgaatc gagctcgact caggggaata tattgacgcg gatctggtta
     3241 tagtgggtat tggtttgctc cccaacgttg accttgcaag cgcagctggc ctcacgtgcg
     3301 aaagtggcat tgtggtagac agtcgttgcc aaacgtccgc acctggaatc tacgcagccg
     3361 gcgattgcac gcaatatcca tcccctatct acggtcggcc acttcatttg gaatctgttc
     3421 ataatgcgat cgaacaggct aagaccgcag cagccgcaat cttggggcgg gatgaaccgt
     3481 ttcgtcaggt tccctggttc tggtcggatc agtacaacat taaacttcag accgcaggcg
     3541 ttaatgaggg ttacgatgat gtcatcattc gcggtgaccc cgcatcagcg agctttgccg
     3601 cattttactt aagagcgggg aaattactgg ctgtggatgc gattaatcgg ccgcgtgagt
     3661 tcatggcttc caaaaccctg attgctgaaa gggccgaagt agacccgacc caattggcag
     3721 acgagagctt gcctccgacc gccctggcgg cagcggtcaa tggtcctact cgggctacga
     3781 gtcccacaag cttataagga tctaggagga taaagaaatg cccaagatta cgtacattga
     3841 ttacacggga acgtcacgct gtgttgatgc tgaaaatggt atgtcattaa tggaaattgc
     3901 gataaacaat aacgttcccg gaattgatgg agattgcggc ggcgagtgcg cgtgtgccac
     3961 gtgtcatgtg catgttgacg ccgattggct ggataaatta cctccttcca gcgaccagga
     4021 ggttagtatg ttggaatttt gtgacggtgt tgatcatact tcccgcttag ggtgtcagat
     4081 taaaatttgc ccgaccttag atggtattgt ggtacggacg cccgccgcgc agcactaagg
     4141 atccaaactc gagtaaggat ctccaggcat caaataaaac gaaaggctca gtcgaaagac
     4201 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag tcacactggc
     4261 tcaccttcgg gtgggccttt ctgcgtttat acctaggcta cagccgatag tctggaacag
     4321 cgcacttacg ggttgctgcg caacccaagt gctaccggcg cggcagcgtg acccgtgtcg
     4381 gcggctccaa cggctcgcca tcgtccagaa aacacggctc atcgggcatc ggcaggcgct
     4441 gctgcccgcg ccgttcccat tcctccgttt cggtcaaggc tggcaggtct ggttccatgc
     4501 ccggaatgcc gggctggctg ggcggctcct cgccggggcc ggtcggtagt tgctgctcgc
     4561 ccggatacag ggtcgggatg cggcgcaggt cgccatgccc caacagcgat tcgtcctggt
     4621 cgtcgtgatc aaccaccacg gcggcactga acaccgacag gcgcaactgg tcgcggggct
     4681 ggccccacgc cacgcggtca ttgaccacgt aggccaacac ggtgccgggg ccgttgagct
     4741 tcacgacgga gatccagcgc tcggccacca agtccttgac tgcgtattgg accgtccgca
     4801 aagaacgtcc gatgagcttg gaaagtgtct tctggctgac caccacggcg ttctggtggc
     4861 ccatctgcgc cacgaggtga tgcagcagca ttgccgccgt gggtttcctc gcaataagcc
     4921 cggcccacgc ctcatgcgct ttgcgttccg tttgcaccca gtgaccgggc ttgttcttgg
     4981 cttgaatgcc gatttctctg gactgcgtgg ccatgcttat ctccatgcgg taggggtgcc
     5041 gcacggttgc ggcaccatgc gcaatcagct gcaacttttc ggcagcgcga caacaattat
     5101 gcgttgcgta aaagtggcag tcaattacag attttcttta acctacgcaa tgagctattg
     5161 cggggggtgc cgcaatgagc tgttgcgtac cccccttttt taagttgttg atttttaagt
     5221 ctttcgcatt tcgccctata tctagttctt tggtgcccaa agaagggcac ccctgcgggg
     5281 ttcccccacg ccttcggcgc ggctccccct ccggcaaaaa gtggcccctc cggggcttgt
     5341 tgatcgactg cgcggccttc ggccttgccc aaggtggcgc tgcccccttg gaacccccgc
     5401 actcgccgcc gtgaggctcg gggggcaggc gggcgggctt cgcccttcga ctgcccccac
     5461 tcgcataggc ttgggtcgtt ccaggcgcgt caaggccaag ccgctgcgcg gtcgctgcgc
     5521 gagccttgac ccgccttcca cttggtgtcc aaccggcaag cgaagcgcgc aggccgcagg
     5581 ccggaggcac tagtgcttgg attctcacca ataaaaaacg cccggcggca accgagcgtt
     5641 ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag gagtccaagc
     5701 gagctctcga accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga
     5761 tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc
     5821 cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca
     5881 cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg
     5941 gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga
     6001 gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat
     6061 cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt
     6121 cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg
     6181 atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca
     6241 atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc
     6301 ccgtcgtggc cagccacgat agccgcgctg cctcgtcctg cagttcattc agggcaccgg
     6361 acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg
     6421 catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag
     6481 cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg
     6541 tctcttgatc agatcatgat cccctgcgcc atcagatcct tggcggcaag aaagccatcc
     6601 agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc aattcc
//
