LOCUS       sequence-127396-        7381 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses E. coli HpaBC in monocistronic form from ePathBrick 
            vector, pETM6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7381)
  AUTHORS   Jones JA, Collins SM, Lachance DM, Vernacchio VR, Koffas MA
  TITLE     ""Optimization of naringenin and p-coumaric acid hydroxylation using
            the native E. coli hydroxylase complex, HpaBC"".
  JOURNAL   Biotechnol Prog. 2015 Oct 21. doi: 10.1002/btpr.2185.
  PUBMED    26488898
REFERENCE   2  (bases 1 to 7381)
  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: "Biotechnol 
            Prog. 2015 Oct 21. doi: 10.1002/btpr.2185."
COMMENT     Sequence Label: pETM6-HpaBC
FEATURES             Location/Qualifiers
     source          1..7381
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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             42..64
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      1683..1730
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1764..1782
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1783..1807
                     /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             1822..1844
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      2413..2460
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2504..2959
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     2801..2822
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(3074..3865,3866..3934))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3697..3716
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3935..4027)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4108..4696
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4597..4616
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4850..4867
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4882..5024
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4950..4969)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5110..5132
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5126..5317)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5889..6971)
                     /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     6933..6952
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6972..7049)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7255..7274
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(7365..7381,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
       61 gagatataca tatgaaacca gaagatttcc gcgccagtac ccaacgtcct ttcaccgggg
      121 aagagtatct gaaaagcctg caggatggtc gcgagatcta tatctatggc gagcgagtga
      181 aagacgtcac cactcatccg gcatttcgta atgcggcagc gtctgttgcc cagctgtacg
      241 acgcactgca caaaccggag atgcaggact ctctgtgttg gaacaccgac accggcagcg
      301 gcggctatac ccataaattc ttccgcgtgg cgaaaagtgc cgacgacctg cgccagcaac
      361 gcgacgccat cgctgagtgg tcacgcctga gctatggctg gatgggccgt accccagact
      421 acaaagccgc tttcggttgc gcactgggcg cgaatccggg cttttacggt cagttcgagc
      481 agaacgcccg taactggtac acccgtattc aggaaactgg cctctacttt aaccacgcga
      541 ttgttaaccc accgatcgat cgtcatttgc cgaccgataa agtgaaagac gtttacatca
      601 agctggaaaa agagactgac gccgggatta tcgtcagcgg tgcgaaagtg gttgccacca
      661 actcggcgct gactcactac aacatgattg gcttcggctc ggcacaagtg atgggcgaaa
      721 acccggactt cgcactgatg ttcgttgcgc caatggatgc cgatggcgtg aaattaatct
      781 cccgcgcctc ttatgagatg gtcgcgggtg ctaccggctc gccatacgac tacccgctct
      841 ccagccgctt cgatgagaac gatgcgattc tggtgatgga taacgtgctg attccatggg
      901 aaaacgtgct gatctaccgc gattttgatc gctgccgtcg ctggacgatg gaaggcggtt
      961 ttgcccgtat gtatccgctg caagcctgtg tgcgcctggc agtgaaatta gacttcatta
     1021 cggcactgct gaaaaaatca ctcgaatgta ccggcaccct ggagttccgt ggtgtgcagg
     1081 ccgatctcgg tgaagtggta gcgtggcgca acaccttctg ggcattgagt gactcgatgt
     1141 gttcagaagc aacgccgtgg gtcaacgggg cttatttacc ggatcatgcc gcactgcaaa
     1201 cctatcgcgt actggcacca atggcctacg cgaagatcaa aaacattatc gaacgcaacg
     1261 ttaccagtgg cctgatctat ctcccttcca gtgcccgtga cctgaataat ccgcagatcg
     1321 accagtatct ggcgaagtat gtgcgcggtt cgaacggtat ggatcacgtc cagcgcatca
     1381 agatcctcaa actgatgtgg gatgctattg gcagcgaatt tggtggtcgt cacgaactgt
     1441 atgaaatcaa ctactccggt agccaggatg agattcgcct gcagtgtctg cgccaggcac
     1501 aaaactccgg caatatggac aagatgatgg cgatggttga tcgctgcctg tcggaatacg
     1561 accaggacgg ctggactgtg ccgcacctgc acaacaacga cgatatcaac atgctggata
     1621 agctgctgaa ataactagtc gcagcttaat taacctaaac tgctgccacc gctgagcaat
     1681 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctaggatcga
     1741 gatcgatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
     1801 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatataca tatgcaatta
     1861 gatgaacaac gcctgcgctt tcgtgacgcg atggccagcc tgtcggcagc ggtaaatatt
     1921 atcaccaccg agggcgacgc cggacaatgc gggattacgg caacggccgt ctgctcggtc
     1981 acggatacac caccgtcgct gatggtgtgc attaacgcca acagtgcgat gaacccggtt
     2041 tttcagggca acggcaagtt gtgcgtcaac gtcctcaacc atgagcagga actgatggca
     2101 cgccacttcg cgggcatgac aggcatggcg atggaagagc gttttagcct ctcatgctgg
     2161 caaaaaggtc cgctggcgca gccggtgcta aaaggttcgc tggccagtct tgaaggtgag
     2221 atccgcgatg tgcaggcaat tggcacacat ctggtgtatc tggtggagat taaaaacatc
     2281 atcctcagtg cagaaggtca tggacttatc tactttaaac gccgtttcca tccggtgatg
     2341 ctggaaatgg aagctgcgat ttaactagtc gcagcttaat taacctaaac tgctgccacc
     2401 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     2461 ctagcgaaag gaggagtcga ctatatccgg attggcgaat gggacgcgcc ctgtagcggc
     2521 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2581 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2641 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2701 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2761 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     2821 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     2881 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     2941 atattaacgt ttacaatttc tggcggcacg atggcatgag attatcaaaa aggatcttca
     3001 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3061 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3121 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3181 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3241 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3301 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3361 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3421 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3481 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3541 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3601 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     3661 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     3721 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     3781 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     3841 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     3901 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tcatgattga
     3961 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4021 aaacaaatag gtcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4081 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4141 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4201 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     4261 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4321 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4381 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4441 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4501 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4561 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4621 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4681 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4741 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     4801 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     4861 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4921 ttcacaccgc atatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     4981 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     5041 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     5101 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     5161 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     5221 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     5281 gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat
     5341 ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta
     5401 ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga
     5461 tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg
     5521 taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc
     5581 agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg
     5641 ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg
     5701 gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca
     5761 ggagcacgat catgctagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     5821 ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg
     5881 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5941 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc
     6001 accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc
     6061 aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc
     6121 gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat gtccgcacca
     6181 acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca
     6241 accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg
     6301 gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga
     6361 tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac
     6421 agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct
     6481 tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc
     6541 ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta
     6601 atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg
     6661 acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat
     6721 ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca
     6781 atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc
     6841 tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc
     6901 accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt
     6961 actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg
     7021 cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc
     7081 ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa
     7141 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     7201 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     7261 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     7321 tccggcgtag cctaggatcg agatcgatct cgatcccgcg aaattaatac gactcactat
     7381 a
//
