LOCUS       sequence-239709-        7449 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plamid was used for biosynthesis of Prenylated 
            4‑Hydroxybenzoate in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7449)
  AUTHORS   He BB, Bu XL, Zhou T, Li SM, Xu MJ, Xu J
  TITLE     Combinatory Biosynthesis of Prenylated 4-Hydroxybenzoate Derivatives
            by Overexpression of the Substrate-Promiscuous Prenyltransferase 
            XimB in Engineered E. coli.
  JOURNAL   ACS Synth Biol. 2018 Sep 21;7(9):2094-2104. doi: 
            10.1021/acssynbio.8b00070. Epub 2018 Aug 30.
  PUBMED    30103600
REFERENCE   2  (bases 1 to 7449)
  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"; doi: 
            "10.1021/acssynbio.8b00070"; journalName: "ACS Synth Biol"; date: 
            "2018-09-21- 21"; volume: "7"; issue: "9"; pages: "2094-2104"
COMMENT     Sequence Label: pXM03
FEATURES             Location/Qualifiers
     source          1..7449
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1116..1138
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1153..1177
                     /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)."
     promoter        complement(1178..1196)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(1226..1273)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1340..1357)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(2345..2377)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     RBS             2385..2407
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2422..2446
                     /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)."
     promoter        complement(2447..2465)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2554..2573)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2779..2856
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2857..3939
                     /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     complement(2876..2895)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4748..4939
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4933..4955)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5041..5180
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5096..5115
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5195..5212)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5366..5954)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5446..5465)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(6065..6084)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             6076..6891
                     /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             6076..6891
                     /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     complement(6149..6168)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(6983..7438)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7120..7141)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7332..7351
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggaact agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtcaaaag actctccccg
      181 caacgatggc gacgagcacg agccacccga agaaacggtt tgccatgaac ttttcccggc
      241 agtcggcagg gctgtcgatg tccaccgtca ccacctgcca gagcaggtgt gccacgcccg
      301 gcagaaacag cgcgtagaac gtccagtgca gatcggcgac ggctccggca gccacgatcc
      361 cgaccacggt cgccccgtaa aacccggcga cccagggccg ggtcgccctg ccgaggcgca
      421 gcgcgaggga cttcaccccc acctgcacgt cgtccgcctt gtcctggtgg gcgtagatcg
      481 tgtcgtaccc cagcgtccag aagacaccgg cgacgaagag caggagcgcg ggagtctcga
      541 tcctcccggc caccgcgagc caacccgcga ggatgtagca gccgaagacc atccccagcc
      601 aggcctgcgg ccagtaggtg atccgcttca tgaacgggta cgcgaccacg agcggatagg
      661 aggcggccac gaacaccgcc gccggaacgc tggccgcagc cagcacgagc agccccgcca
      721 ccgcctgcac cacggcgaag acaagcgctc ccgtgaccgt gaccgtaccg gccacgagcg
      781 gcctcccgac ggtacgtgcc acccgggcgt cgaacttgcg gtcggcaagg tcgttgacga
      841 cgcagccgaa gccgcgcacc agcaccccac cgaccgtgaa cagcagcacc tggcgccagt
      901 cgggcagccc cgccgaggcg agcgcgatcc cccacaggcc ggggaggagg tacagccaac
      961 tgcctatggg cgcgtgcata cgcgccagtt gtagataggg gaggacggcc ttcggcgcac
     1021 tcggcacgac cgacgccgcg atgctcggca cgacgaaccg atatcgcgtg ctctgctgcg
     1081 agggttgagc ggaaatatcg atcaccatat gtatatctcc ttcttaaagt taaacaaaat
     1141 tatttgtaga ggggaattgt tatccgctca caattcccct atagtgagtc gtattaattt
     1201 cgcggtttct agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
     1261 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
     1321 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagctttt
     1381 aattctgacg agatgcaatg tagtctgcta acgcaaatag aggtgctgct ttggtaggat
     1441 caaaacaact aagttcctgt tttccacggt tcagtaattc atcggcaaat tccttcgctt
     1501 tttccaaacc catgagtttg ggataggtgg ctttgtcaga aatcaaatct ttcccagcag
     1561 tctttccgag ttcttccgag gactggctta catccaaaat atcatcgaca acttggaaaa
     1621 gcaatcctac acagcgagcg taccgtctag cacgctcgat gtcgtcctcg ctggcacccc
     1681 ccataattgc gccacacacg acactgcatt ccaacaacac agccgtttta tgaatatgga
     1741 cccattccag cgtttctaag tctatactgg gatcaccttc gctttcaata tccaccactt
     1801 gaccacccat cacgccctca cttcctgtgg cgcgtccaat ttcggatagc aaccgtaaaa
     1861 taatgtcagt acctagggta cgactggtgc tcacggctac atgttcaaag gccaatgaca
     1921 ataatgcatc gccagcaatg atcgccgtgt cttcaccaaa aactttgtgg ttggtaggct
     1981 taccgcgacg caaatcatcg ttatcaatat agggcaaatc gtcatgaatc aaactcatag
     2041 tgtgaatcat ctcgatggca caagccgtag gcatggcaag ttcctcagtc ccccccatta
     2101 gctcacaggc cgcaatacat aaaattggtc gaaccctctt cccgccggct aggagggaat
     2161 agcgcatact ttcatagatt ttttgaggat agcggggggg aataacttta tctaaggcct
     2221 cattaaccgc aatggccttg gagtgcatat acttgtcaaa atcaaattcc accacgtttt
     2281 ttacttgttc ggatagatca gctagttgga ccaaggcctt actgctgcgc gtcatcggat
     2341 cccgacccat ttgctgtcca ccagtcatgc tagccatatg tatatctcct tcttaaagtt
     2401 aaacaaaatt atttgtagag gggaattgtt atccgctcac aattccccta tagtgagtcg
     2461 tattaatttc gcggtttcta gagcagatct cgatcctcta cgccggacgc atcgtggccg
     2521 gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc accgatgggg
     2581 aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt atggtggcag
     2641 gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc cttgcggcgg
     2701 cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag tcgcataagg
     2761 gagagcgtcg agatcccgga caccatcgaa tggcgcaaaa cctttcgcgg tatggcatga
     2821 tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac gttatacgat
     2881 gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc
     2941 cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt
     3001 cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc
     3061 tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat
     3121 caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa
     3181 gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg
     3241 gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt
     3301 gatgtctctg accagacacc catcaacagt attattttct cccatgaaga cggtacgcga
     3361 ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca
     3421 ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat
     3481 caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa
     3541 accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag
     3601 atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc
     3661 tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc gttaaccacc
     3721 atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct
     3781 cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa aagaaaaacc
     3841 accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     3901 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag
     3961 ttagctcact cattaggcac cgggatctcg accgatgccc ttgagagcct tcaacccagt
     4021 cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt
     4081 tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg gcgaggaccg
     4141 ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc
     4201 cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga agcaggccat
     4261 tatcgccggc atggcggccc cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg
     4321 gctaggctgg cggggttgcc ttactggtta gcagaatgaa tcaccgatac gcgagcgaac
     4381 gtgaagcgac tgctgctgca aaacgtctgc gacctgagca acaacatgaa tggtcttcgg
     4441 tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca ttatgttccg
     4501 gatctgcatc gcaggatgct gctggctacc ctgtggaaca cctacatctg tattaacgaa
     4561 gcgctggcat tgaccctgag tgatttttct ctggtcccgc cgcatccata ccgccagttg
     4621 tttaccctca caacgttcca gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag
     4681 catcctctct cgtttcatcg gtatcattac ccccatgaac agaaatcccc cttacacgga
     4741 ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca
     4801 gacattaacg cttctggaga aactcaacga gctggacgcg gatgaacagg cagacatctg
     4861 tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc gtttcggtga
     4921 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     4981 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     5041 cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca
     5101 tcagagcaga ttgtactgag agtgcaccat tgcggtgtga aataccgcac agatgcgtaa
     5161 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5221 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5281 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5341 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5401 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5461 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5521 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5581 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5641 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5701 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5761 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     5821 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5881 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5941 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6001 aaaactcacg ttaagggatt ttggtcatga acaataaaac tgtctgctta cataaacagt
     6061 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cttgctctag gccgcgatta
     6121 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     6181 tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa
     6241 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     6301 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     6361 ttactcacca ctgcgatccc cgggaaaaca gcattccagg tattagaaga atatcctgat
     6421 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     6481 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     6541 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     6601 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     6661 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     6721 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     6781 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     6841 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta agaattaatt
     6901 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6961 atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta
     7021 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     7081 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     7141 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     7201 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     7261 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     7321 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     7381 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     7441 cattcgcca
//
