LOCUS       sequence-379998-        6120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Full length wt AtPCO2 with Nt-His tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6120)
  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 6120)
  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)-AtPCO2(wt)
FEATURES             Location/Qualifiers
     source          1..6120
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     415..434
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     668..685
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    700..839
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(765..784)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     925..947
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(941..1132)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1941..3023)
                     /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     2985..3004
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3024..3101)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3307..3326
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3410..3429
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3410..3428
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3429..3453
                     /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             3468..3490
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3509..3526
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3536..3553
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             4394..4411
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4464..4482)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4478..4525
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4562..5017
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4649..4668)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4859..4880
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5109..5924)
                     /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(5109..5924)
                     /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     5832..5851
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5916..5935
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(6046..6120,1..514)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
       61 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
      121 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
      181 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
      241 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
      301 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
      361 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
      421 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
      481 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
      541 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
      601 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
      661 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
      721 tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat
      781 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
      841 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
      901 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
      961 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
     1021 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     1081 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct
     1141 ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc
     1201 tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac
     1261 aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct
     1321 tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc
     1381 ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac
     1441 cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg
     1501 ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc
     1561 gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg ccggcgataa
     1621 tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg
     1681 cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc
     1741 ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa
     1801 agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga
     1861 ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac
     1921 ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     1981 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg
     2041 gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga
     2101 gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg
     2161 gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag
     2221 atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc
     2281 tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt
     2341 tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga
     2401 ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat
     2461 gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt
     2521 cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca
     2581 agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc
     2641 agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct
     2701 ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga
     2761 tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag
     2821 gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga
     2881 atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg
     2941 ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca
     3001 tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat
     3061 catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc
     3121 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc
     3181 cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
     3241 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     3301 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     3361 ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc
     3421 actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt
     3481 aagaaggaga tataccatgg gcagcagcca tcatcatcat catcacagca gcggcctggt
     3541 gccgcgcggc agccatatgg gaactgatac agttatgtct ggacgagtaa ggaaagatct
     3601 atcaaaaacg aatccaaatg ggaatattcc tgagaatcgt tctaattcac gtaagaagat
     3661 ccaacgacgg agtaagaaaa ccctaatctg tccggttcaa aaactattcg atacttgtaa
     3721 gaaagttttc gctgatggca aatctggtac cgtcccttct caagaaaaca ttgagatgct
     3781 tcgagccgtt ttggatgaaa tcaagcctga ggatgttggc gtaaatccta agatgtcgta
     3841 ttttcgatct acagtgaccg gacgatctcc gttagtgacg tatcttcaca tctatgcatg
     3901 tcatagattc tcgatttgca ttttctgttt acctccatct ggtgttatcc ctcttcacaa
     3961 tcacccggag atgactgtgt ttagtaagct cttgtttggt acaatgcata tcaaatccta
     4021 tgattgggtc cctgattctc cccagccgag ttcagatact cgtttggcga aagtgaaagt
     4081 agattcggac tttaccgcac cttgtgatac ttctatactg tacccggctg atggagggaa
     4141 tatgcattgc ttcaccgcga aaacggcttg cgcggttctt gatgttattg gtcctccata
     4201 ctctgatccc gcaggacgtc attgtactta ctatttcgat tatccgttct ctagtttctc
     4261 ggtcgatgga gtcgtggttg ccgaggagga gaaggaaggc tatgcatggt tgaaggagag
     4321 ggaagagaag ccagaggatt taacggttac tgcattgatg tatagtggac caaccatcaa
     4381 agaatgactc gagcaccacc accaccacca ctgagatccg gctgctaaca aagcccgaaa
     4441 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     4501 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg
     4561 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4621 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4681 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4741 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4801 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4861 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4921 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4981 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     5041 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5101 attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     5161 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     5221 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
     5281 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     5341 gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca
     5401 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
     5461 cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca
     5521 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     5581 tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac
     5641 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     5701 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     5761 ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat
     5821 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     5881 aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta
     5941 ctgtttatgt aagcagacag ttttattgtt catgaccaaa atcccttaac gtgagttttc
     6001 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6061 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
//
