LOCUS       sequence-320871-        7296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of aryl-alcohol dehydrogenase from the white-rot fungus 
            Phanerochaete chrysosporium.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7296)
  AUTHORS   Yang DD, Francois JM, de Billerbeck GM
  TITLE     Cloning, expression and characterization of an aryl-alcohol 
            dehydrogenase from the white-rot fungus Phanerochaete chrysosporium 
            strain BKM-F-1767.
  JOURNAL   BMC Microbiol. 2012 Jun 28;12:126. doi: 10.1186/1471-2180-12-126.
  PUBMED    22742413
REFERENCE   2  (bases 1 to 7296)
  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: "BMC 
            Microbiol."; date: "2012-06-28"; volume: "12:126. doi"; pages: " 
            10.1186/1471-2180-12-126"
COMMENT     Sequence Label: pGS-21a-PcAad1
FEATURES             Location/Qualifiers
     source          1..7296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             101..118
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             128..781
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             818..832
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             2024..2041
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2094..2112)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2108..2155
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2192..2647
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2279..2298)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2489..2510
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2673..2777
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2778..2846,2847..3638)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2996..3015)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3809..4397
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4298..4317
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4551..4568
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4583..4722
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4648..4667)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4808..4830
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4824..5015)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5824..6906)
                     /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     6868..6887
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6907..6984)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7190..7209
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gtctggttct catcatcatc atcatcatag
      121 cagcggtatg tcccctatac taggttattg gaaaattaag ggccttgtgc aacccactcg
      181 acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg
      241 tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc ttccttatta
      301 tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata tagctgacaa
      361 gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc
      421 ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact ttgaaactct
      481 caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag atcgtttatg
      541 tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt tgtatgacgc
      601 tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa aattagtttg
      661 ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat ccagcaagta
      721 tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc atcctccaaa
      781 atcggatctg ggccacacag gccatagatc tggtaccgac gacgacgaca aggccatggc
      841 tgatatcgga tccatgaaca tctgggcacc cgcacctgag cctccgacta agctcggccg
      901 tcacaggcag cttgctcccg gatgtggtct ccatgtatcg cccatccagc tcggcgcaat
      961 gagtatcggc gacaagtggc atccatatgg catgggcacc atggacaagg aagccagctt
     1021 caagctcctc gacgcattct acaacgctgg tggtaacttc attgataccg ctaatgtcta
     1081 ccaagacgag acatccgagg aatttattgg cgagtggatg gaggctcggg gcaaccgcga
     1141 ccagatggtc gttgctacca agtacagtct ggtctacaaa cggggtgcca gcttcgaaga
     1201 gatcccgcag aagacgcagt atgtcggcaa cagcctcaag tcgatgcaca tctcagtgca
     1261 cgactcattg cgcaagctgc gcacgagcta catcgatata ttctacgttc acttctggga
     1321 ctacacttgc acaattgagg aggtgatgaa cggcttgcac aatcttgttg cacagggcaa
     1381 ggttctctac ctcggtgtct cggacacacc cgcctgggtt gtctcgaagg cgaacaacta
     1441 tgcgcgcatg gccggcaaga cgcctttcgt catctacgag ggagagtgga acatcacgat
     1501 gcgcgacatg gagcgcgaca tcattccgat gtgcattcac gaagggatgg ccatcgcgcc
     1561 ctggaatgtg ctttgcgccg gcaagatacg cacggacgcc gaggaggagc gccgtctcaa
     1621 gtccggcgag ggtgggcgta cgctcctaca gttcgacgga tggctccgga acgagaccga
     1681 gcgcaaggtc agcaaggcac tcgagaaggt cgctgaagag atcggcgcaa agagtatcac
     1741 atccgtcgca attgcgtacc tcatgcagaa gttcccctac gtcttcccca ttgtcggtgg
     1801 acgcaaggtc gaacacctgt acgccaacct cgaggcgctc gacatctcgc tcagccccga
     1861 gcagatgcag ttcctgaacg acacagtacc cttcaacaag ggcttcccat accttctctt
     1921 cggtgatggg tccgactaca acatcgtaca caaggctgca ggtcattatg acaagtggcc
     1981 cgcgcaacag gctatccgcc cccagaaggc ggccgcactc gagcaccacc accaccacca
     2041 ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     2101 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     2161 aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     2221 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     2281 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     2341 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     2401 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     2461 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     2521 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     2581 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     2641 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2701 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2761 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2821 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2881 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2941 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     3001 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     3061 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     3121 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3181 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3241 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3301 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3361 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3421 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3481 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3541 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3601 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3661 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3721 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3781 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3841 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3901 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3961 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4021 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4081 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4141 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4201 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4261 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4321 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4381 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4441 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4501 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4561 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4621 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4681 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     4741 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4801 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     4861 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     4921 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     4981 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     5041 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     5101 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     5161 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     5221 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     5281 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     5341 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     5401 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
     5461 agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg
     5521 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat
     5581 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg
     5641 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
     5701 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     5761 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
     5821 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5881 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     5941 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     6001 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     6061 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg
     6121 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     6181 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     6241 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     6301 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     6361 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     6421 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     6481 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     6541 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     6601 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     6661 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     6721 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     6781 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     6841 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     6901 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     6961 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca
     7021 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     7081 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     7141 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     7201 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     7261 cgtagaggat cgagatcgat ctcgatcccg cgaaat
//
