LOCUS       sequence-173997-        6636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses PvdA, the ornithine hydroxylase from Pseudomonas 
            aeruginosa.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6636)
  AUTHORS   Olucha J, Meneely KM, Chilton AS, Lamb AL
  TITLE     Two structures of an N-hydroxylating flavoprotein monooxygenase: 
            ornithine hydroxylase from Pseudomonas aeruginosa.
  JOURNAL   J Biol Chem. 2011 Sep 9;286(36):31789-98. doi: 
            10.1074/jbc.M111.265876. Epub 2011 Jul 13.
  PUBMED    21757711
REFERENCE   2  (bases 1 to 6636)
  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: 
            "10.1074/jbc.M111.265876"; journalName: "J Biol Chem"; date: 
            "2011-09-9- 9"; volume: "286"; issue: "36"; pages: "31789-98"
COMMENT     Sequence Label: pET28b-pvdA WT
FEATURES             Location/Qualifiers
     source          1..6636
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             517..534
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(587..605)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      601..648
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      685..1140
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(772..791)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     982..1003
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1232..2047)
                     /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             complement(1232..2047)
                     /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     1955..1974
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2039..2058
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2169..2757
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2658..2677
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2911..2928
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2943..3082
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3008..3027)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3168..3190
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3184..3375)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4184..5266)
                     /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     5228..5247
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5267..5344)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5550..5569
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5653..5672
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5653..5671
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5672..5696
                     /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             5711..5733
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5752..5769
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5779..5796
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
ORIGIN
        1 taccacaaca ccaactattc ggtggtggat accgacctga tcgagcgcat ctacggcgtc
       61 ttctaccgcc agaaagtctc cggcatcccg cgccacgcct tccgttgcat gaccaccgtg
      121 gagcgcgcga ccgccaccgc ccagggcatc gagctggcgt tgcgcgacgc cggtagcggc
      181 gagctaagcg tagagaccta cgacgcagtg atcctggcca ccggctatga gcgccagttg
      241 caccgccaac tgctcgaacc gctggcggag tacctcggcg accacgagat cggccgcgac
      301 taccgcctgc agaccgacga gcgctgcaag gtggcgatct acgcgcaggg cttcagccag
      361 gccagccatg gcctcagcga caccctgctg tcggtgctgc cggtgcgtgc cgaggaaatc
      421 tccggctccc tgtaccagca cctgaagccc ggtactgcgg cccgcgccct gcacgagcac
      481 gccctggcca gctgaaagct tgcggccgca ctcgagcacc accaccacca ccactgagat
      541 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
      601 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
      661 actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
      721 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
      781 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
      841 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
      901 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
      961 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     1021 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     1081 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     1141 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     1201 ttcaaatatg tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa
     1261 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
     1321 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
     1381 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
     1441 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat
     1501 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
     1561 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc
     1621 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
     1681 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc
     1741 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
     1801 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
     1861 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     1921 atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     1981 aatcagcatc catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat
     2041 ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgacc
     2101 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     2161 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     2221 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2281 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     2341 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2401 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     2461 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     2521 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2581 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2641 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2701 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     2761 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     2821 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     2881 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     2941 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca
     3001 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
     3061 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     3121 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     3181 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     3241 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     3301 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     3361 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
     3421 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
     3481 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
     3541 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
     3601 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
     3661 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
     3721 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
     3781 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg
     3841 tggggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc
     3901 agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat
     3961 catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac
     4021 ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc
     4081 ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc
     4141 cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca
     4201 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4261 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
     4321 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
     4381 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg
     4441 tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg
     4501 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
     4561 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
     4621 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
     4681 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
     4741 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
     4801 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
     4861 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
     4921 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
     4981 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
     5041 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
     5101 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
     5161 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
     5221 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga
     5281 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg
     5341 tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
     5401 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc
     5461 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
     5521 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     5581 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg
     5641 atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac aattcccctc
     5701 tagaaataat tttgtttaac tttaagaagg agatatacca tgggcagcag ccatcatcat
     5761 catcatcaca gcagcggcct ggtgccgcgc ggcagccata tgactcaggc aactgcaacc
     5821 gccgtggttc acgatctcat cggtgtcggc ttcggccctt ccaatatcgc cctggcgatt
     5881 gccctccagg aacgggccca ggcgcagggc gccctggaag tgctgttcct ggacaagcag
     5941 ggcgactacc gctggcacgg caacaccctg gtgtcgcaga gcgagttgca gatctccttc
     6001 ctcaaggacc tggtgtccct gcgcaacccc accagtccgt attccttcgt caactacctg
     6061 cacaagcacg atcgtctggt cgacttcatc aacctgggca ccttctatcc ctgccggatg
     6121 gagttcaacg actacctgcg ctgggtcgcc agccacttcc aggagcagag ccgctacggc
     6181 gaagaggtcc tgcgcatcga gccgatgctg agcgccggcc aggtcgaggc gctgcgggtg
     6241 atctcgcgca acgccgacgg cgaggagctg gtgcgcacca cccgcgccct ggtggtcagt
     6301 cccggcggca ccccgcgtat cccgcaggtg ttccgtgcgc tcaagggcga cggccgggtg
     6361 ttccaccaca gccagtacct ggaacacatg gccaagcagc cctgcagcag tggcaagccg
     6421 atgaagatcg ccattatcgg cggcgggcag agcgcggcgg aggccttcat cgacctcaac
     6481 gacagctacc cgtcggtgca ggccgacatg atcctgcgtg cctcggcgct caagccggcg
     6541 gacgatagcc cgttcgtcaa cgaagtgttc gcgccgaagt tcaccgatct catctacagc
     6601 cgcgaacatg ccgaacgcga gcgtttgctg cgcgaa
//
