LOCUS       sequence-380001-        6041 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Full length wt AtPCO4 with Nt-His tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6041)
  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 6041)
  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-AtPCO4-1(wt)
FEATURES             Location/Qualifiers
     source          1..6041
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     153..172
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     406..423
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    438..580
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(506..525)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     666..688
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(682..873)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1682..2764)
                     /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     2726..2745
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2765..2842)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3048..3067
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3151..3170
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3151..3169
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3170..3194
                     /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             3209..3231
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3250..3267
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3292..3312
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             4052..4069
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4122..4140)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4136..4183
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4220..4675
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4307..4326)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4517..4538
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4768..5583)
                     /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(4768..5583)
                     /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     5491..5510
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5575..5594
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(5705..6041,1..252)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
       61 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
      121 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
      181 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
      241 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
      301 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
      361 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
      421 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
      481 caccgcatat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
      541 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
      601 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
      661 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
      721 gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc
      781 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
      841 catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct
      901 gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga
      961 tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg
     1021 gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg
     1081 tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg
     1141 cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt
     1201 tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga
     1261 ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag
     1321 cacgatcatg cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa
     1381 acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac
     1441 cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac
     1501 ccagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc
     1561 ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa
     1621 gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
     1681 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     1741 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
     1801 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
     1861 ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat
     1921 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca
     1981 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
     2041 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
     2101 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
     2161 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
     2221 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
     2281 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
     2341 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
     2401 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
     2461 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
     2521 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
     2581 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
     2641 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
     2701 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
     2761 tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     2821 ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt
     2881 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
     2941 tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
     3001 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
     3061 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
     3121 tagaggatcg agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag
     3181 cggataacaa ttcccctcta gaaataattt tgtttaactt taagaaggag atataccatg
     3241 ggcagcagcc atcatcatca tcatcactct tctggtgtag atctgggtac cgagaacctg
     3301 tacttccaat ccaacgcgat gccttacttt gctcagaggc tttacaatac ttgcaaggcg
     3361 tctttctctt cagacggacc aataacagaa gatgctctgg agaaggttcg caatgtcttg
     3421 gagaaaatca agccatctga tgttggaatt gagcaggatg ctcaattggc acggtccagg
     3481 tctggtcctc tcaatgaacg caatggaagt aatcagtctc ccccagcaat aaagtatctt
     3541 catttgcatg agtgtgacag tttctctata ggaatcttct gtatgccacc ttcttctatg
     3601 atacctcttc ataaccatcc gggcatgacc gtgctaagca agctcgttta tggttcaatg
     3661 catgtgaagt catatgattg gctagagcct caactgaccg aaccagagga tccatcacaa
     3721 gcaagacctg ctaaactggt gaaggatact gagatgacgg ctcaaagccc agtaacaaca
     3781 ttatatccga aaagtggtgg caacattcac tgtttcaaag ccatcaccca ttgtgctatt
     3841 cttgacatct tagctccacc ttactcttca gagcatgatc ggcattgcac ttacttccgg
     3901 aaatccagaa gagaagactt accaggtgaa ttagaagtgg atggagaagt ggtcacagac
     3961 gtgacatggc ttgaggaatt tcaaccgcct gatgactttg tgatacgacg aattccgtac
     4021 agaggtcctg tcattagaac ttgaactcga gcaccaccac caccaccact gagatccggc
     4081 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     4141 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     4201 atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4261 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4321 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4381 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4441 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4501 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4561 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4621 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttcaggt
     4681 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4741 aatatgtatc cgctcatgaa ttaattctta gaaaaactca tcgagcatca aatgaaactg
     4801 caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga
     4861 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
     4921 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
     4981 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat
     5041 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc
     5101 aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt
     5161 aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc
     5221 aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg
     5281 gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg
     5341 aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc
     5401 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg
     5461 atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
     5521 agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct
     5581 cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc atgaccaaaa
     5641 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5701 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5761 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5821 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     5881 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5941 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6001 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac a
//
