LOCUS       sequence-467735-        6044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Full length wt AtPCO4-2 with Nt-His tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6044)
  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 6044)
  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-AtPCO4-2(wt)
FEATURES             Location/Qualifiers
     source          1..6044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(36..1118)
                     /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     1080..1099
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1119..1196)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1402..1421
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1505..1524
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1505..1523
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1524..1548
                     /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             1563..1585
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1604..1621
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1646..1666
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2409..2426
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2479..2497)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2493..2540
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2577..3032
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2664..2683)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2874..2895
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3125..3940)
                     /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(3125..3940)
                     /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     3848..3867
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3932..3951
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4062..4650
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4551..4570
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4804..4821
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4836..4978
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4904..4923)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5064..5086
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5080..5271)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
       61 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
      121 ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc
      181 accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga
      241 aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg
      301 tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt
      361 gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc
      421 agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct
      481 atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc
      541 gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga
      601 tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc
      661 tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg
      721 gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga
      781 ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg
      841 ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc
      901 agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt
      961 gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt
     1021 ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg
     1081 gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc
     1141 tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg
     1201 atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
     1261 gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
     1321 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
     1381 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     1441 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg
     1501 aaattaatac gactcactat aggggaattg tgagcggata acaattcccc tctagaaata
     1561 attttgttta actttaagaa ggagatatac catgggcagc agccatcatc atcatcatca
     1621 ctcttctggt gtagatctgg gtaccgagaa cctgtacttc caatccaacg cgatgcctta
     1681 ctttgctcag aggctttaca atacttgcaa ggcgtctttc tcttcagacg gaccaataac
     1741 agaagatgct ctggagaagg ttcgcaatgt cttggagaaa atcaagccat ctgatgttgg
     1801 aattgagcag gatgctcaat tggcacggtc caggtctggt cctctcaatg aacgcaatgg
     1861 aagtaatcag tctcccccag caataaagta tcttcatttg catgagtgtg acagtttctc
     1921 tataggaatc ttctgtatgc caccttcttc tatgatacct cttcataacc atccgggcat
     1981 gaccgtgcta agcaagctcg tttatggttc aatgcatgtg aagtcatatg attggctaga
     2041 gcctcaactg accgaaccag aggatccatc acaagaagca agacctgcta aactggtgaa
     2101 ggatactgag atgacggctc aaagcccagt aacaacatta tatccgaaaa gtggtggcaa
     2161 cattcactgt ttcaaagcca tcacccattg tgctattctt gacatcttag ctccacctta
     2221 ctcttcagag catgatcggc attgcactta cttccggaaa tccagaagag aagacttacc
     2281 aggtgaatta gaagtggatg gagaagtggt cacagacgtg acatggcttg aggaatttca
     2341 accgcctgat gactttgtga tacgacgaat tccgtacaga ggtcctgtca ttagaacttg
     2401 aactcgagca ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag
     2461 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     2521 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc
     2581 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2641 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2701 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2761 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2821 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2881 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2941 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3001 gaattttaac aaaatattaa cgtttacaat ttcaggtggc acttttcggg gaaatgtgcg
     3061 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta
     3121 attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     3181 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     3241 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     3301 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     3361 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag
     3421 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     3481 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     3541 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     3601 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     3661 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     3721 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     3781 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     3841 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     3901 gcggcctaga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     3961 ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc
     4021 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4081 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4141 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4201 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4261 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4321 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4381 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4441 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4501 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4561 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4621 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4681 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4741 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4801 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     4861 catctgtgcg gtatttcaca ccgcatatat ggtgcactct cagtacaatc tgctctgatg
     4921 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     4981 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     5041 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     5101 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     5161 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     5221 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
     5281 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
     5341 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     5401 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
     5461 cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg
     5521 gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc
     5581 gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt
     5641 tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg
     5701 ggtcctcaac gacaggagca cgatcatgcg cacccgtggg gccgccatgc cggcgataat
     5761 ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc
     5821 gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg
     5881 gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa
     5941 gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac
     6001 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcct
//
