LOCUS       sequence-380005-        6046 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Full length wt AtPCO5 with Nt-His tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6046)
  AUTHORS   White MD, Kamps JJAG, East S, Taylor Kearney LJ, Flashman E
  TITLE     The plant cysteine oxidases from Arabidopsis thaliana are 
            kinetically tailored to act as oxygen sensors.
  JOURNAL   J Biol Chem. 2018 Jul 27;293(30):11786-11795. doi: 
            10.1074/jbc.RA118.003496. Epub 2018 May 30.
  PUBMED    29848548
REFERENCE   2  (bases 1 to 6046)
  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.RA118.003496"; journalName: "J Biol Chem"; date: 
            "2018-07-27- 27"; volume: "293"; issue: "30"; pages: "11786-11795"
COMMENT     Sequence Label: pET-28a(+)-His(6)-TEV-AtPCO5(wt)
FEATURES             Location/Qualifiers
     source          1..6046
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    418..560
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(486..505)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     646..668
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(662..853)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1662..2744)
                     /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     2706..2725
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2745..2822)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3028..3047
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3131..3150
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3131..3149
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3150..3174
                     /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             3189..3211
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3232..3249
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3274..3294
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             4037..4054
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4107..4125)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4121..4168
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4205..4660
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4292..4311)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4502..4523
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4753..5568)
                     /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(4753..5568)
                     /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     5476..5495
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5560..5579
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(5690..6046,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
      421 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat atggtgcact
      481 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
      541 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
      601 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
      661 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
      721 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
      781 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
      841 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
      901 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
      961 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
     1021 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
     1081 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
     1141 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
     1201 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
     1261 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
     1321 gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
     1381 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
     1441 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
     1501 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     1561 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
     1621 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     1681 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     1741 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
     1801 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
     1861 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta
     1921 tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg
     1981 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
     2041 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
     2101 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
     2161 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
     2221 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
     2281 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
     2341 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
     2401 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
     2461 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
     2521 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
     2581 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
     2641 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
     2701 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
     2761 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg
     2821 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag
     2881 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa
     2941 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
     3001 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     3061 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat
     3121 cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta
     3181 gaaataattt tgtttaactt taagaaggag atatacccca tgggcagcag ccaccaccac
     3241 caccaccata gcagcggcgt tgacctgggc accgagaatc tgtatttcca aagcaatgcg
     3301 atgccgtatt ttatccaacg tctgttcaac acctgcaaga gcagcctgag cccgaacggt
     3361 ccggtgagcg aggaagcgct ggacaaagtg cgtaacgttc tggagaagat caaaccgagc
     3421 gatgtgggtc tggagcagga agcgcaactg gttcgtaact ggccgggtcc gggtaacgag
     3481 cgtaacggca accaccacag cctgccggcg attaagtacc tgcagctgca cgaatgcgac
     3541 agcttcagca tcggtatctt ctgcatgccg ccgggcagca tcattccgct gcacaaccac
     3601 ccgggtatga ccgtgctgag caagctggtt tacggcagca tgcacgtgaa aagctatgac
     3661 tgggcggagc cggatcagag cgaactggat gatccgctgc aagcgcgtcc ggcgaagctg
     3721 gttaaagaca tcgatatgac cagcccgagc ccggcgacca ccctgtaccc gaccaccggt
     3781 ggcaacatcc actgcttcaa ggcgattacc cactgcgcga tctttgacat tctgagcccg
     3841 ccgtacagca gcacccacgg ccgtcactgc aactattttc gtaaaagccc gatgctggat
     3901 ctgccgggcg agatcgaagt tatgaacggc gaagtgatta gcaacgttac ctggctggag
     3961 gaatatcaac cgccggacaa tttcgttatt tggcgtgtgc cgtatcgtgg cccggtgatt
     4021 cgcaagtaaa ctcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg
     4081 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     4141 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa
     4201 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4261 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4321 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4381 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4441 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4501 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4561 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4621 tttaacgcga attttaacaa aatattaacg tttacaattt caggtggcac ttttcgggga
     4681 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4741 atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc
     4801 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc
     4861 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac
     4921 atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc
     4981 atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg
     5041 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt
     5101 cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca
     5161 aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc
     5221 tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag
     5281 taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc
     5341 cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc
     5401 atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc
     5461 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga
     5521 atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcataa caccccttgt
     5581 attactgttt atgtaagcag acagttttat tgttcatgac caaaatccct taacgtgagt
     5641 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5701 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5761 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5821 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5881 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5941 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6001 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaac
//
