LOCUS       sequence-404766-        5428 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For recombinant protein production in Escherichia coli. His-tagged 
            Arabidopsis thaliana HEME OXYGENASE 1 (HO1) without the plastid 
            transit peptide under the control of the T7 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5428)
  AUTHORS   Ishikawa K, Xie X, Osaki Y, Miyawaki A, Numata K, Kodama Y
  TITLE     Bilirubin is produced nonenzymatically in plants to maintain 
            chloroplast redox status.
  JOURNAL   Sci Adv. 2023 Jun 9;9(23):eadh4787. doi: 10.1126/sciadv.adh4787. 
            Epub 2023 Jun 7.
  PUBMED    37285441
REFERENCE   2  (bases 1 to 5428)
  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"; journalName: "Sci Adv."; 
            date: "2023-06-9"; volume: "9(23):eadh4787. doi"; pages: " 
            10.1126/sciadv.adh4787. Epub 2023 Jun 7"
COMMENT     Sequence Label: pDEST17-HO1(51-231)
FEATURES             Location/Qualifiers
     source          1..5428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             71..93
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             113..130
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    140..164
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(897..921)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(974..992)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      988..1035
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1401..1406,1407..1423,1424..1429))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1456..1474)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1542..1646
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1647..1715,1716..2507)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1865..1884)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2678..3266
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3167..3186
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3420..3437
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3452..3591
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3517..3536)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3677..3699
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3693..3884)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5346..5365
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 agatctcgat cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc
       61 tagaaataat tttgtttaac tttaagaagg agatatacat atgtcgtact accatcacca
      121 tcaccatcac ctcgaatcaa caagtttgta caaaaaagca ggctttaaag gaaccaattc
      181 agtcgactta gtggtggttg cggctactac tgcggcagag aagcagaaga agaggtatcc
      241 tggagaatca aagggttttg tggaggagat gaggtttgtg gctatgagac ttcatactaa
      301 agatcaagct aaggaaggtg agaaagagac taaatctatt gaggaacgtc ctgttgctaa
      361 atgggaacct actgttgaag gttacttgag gtttcttgtg gatagtaaat tggtttatga
      421 tactcttgaa ctgattattc aagactccaa tttcccaact tatgccgagt tcaagaacac
      481 ggggctggaa agagcggaga aattatccac ggatttggag tggttcaagg aacaaggtta
      541 cgagattcca gaaccaacag ctcctggtaa aacatattct caatatttaa aggaattagc
      601 agagaaggat cctcaagcat tcatttgtca cttctacaac atctactttg ctcatagtgc
      661 tggtggacga atgattggca gaaaggtggc agagcggata ctcgataata aagaactcga
      721 gttctacaaa tgggacggcg aactttctca attgttgcag aacgttaggg agaaactgaa
      781 caaggttgca gaggagtgga ctagagaaga aaagaatcat tgtttggaag agactgagaa
      841 atcgttcaag tattctggtg agatacttcg tctcatattg tcctgaggat atctagaccc
      901 agctttcttg tacaaagtgg ttgattcgag gctgctaaca aagcccgaaa ggaagctgag
      961 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     1021 ttgaggggtt ttttgctgaa aggaggaact atatccggat atccacagga cgggtgtggt
     1081 cgccatgatc gcgtagtcga tagtggctcc aagtagcgaa gcgagcagga ctgggcggcg
     1141 gccaaagcgg tcggacagtg ctccgagaac gggtgcgcat agaaattgca tcaacgcata
     1201 tagcgctagc agcacgccat agtgactggc gatgctgtcg gaatggacga tatcccgcaa
     1261 gaggcccggc agtaccggca taaccaagcc tatgcctaca gcatccaggg tgacggtgcc
     1321 gaggatgacg atgagcgcat tgttagattt catacacggt gcctgactgc gttagcaatt
     1381 taactgtgat aaactaccgc attaaagctt atcgatgata agctgtcaaa catgagaatt
     1441 cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     1501 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     1561 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     1621 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     1681 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     1741 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     1801 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     1861 ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc
     1921 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1981 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     2041 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     2101 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2161 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     2221 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     2281 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     2341 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     2401 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     2461 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     2521 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     2581 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     2641 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     2701 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     2761 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     2821 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     2881 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     2941 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3001 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3061 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3121 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3181 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3241 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3301 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3361 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3421 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     3481 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata
     3541 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     3601 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3661 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3721 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     3781 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     3841 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     3901 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
     3961 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
     4021 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
     4081 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     4141 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     4201 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     4261 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
     4321 aacgacagga gcacgatcat gcgcacccgt ggccaggacc caacgctgcc cgagatgcgc
     4381 cgcgtgcggc tgctggagat ggcggacgcg atggatatgt tctgccaagg gttggtttgc
     4441 gcattcacag ttctccgcaa gaattgattg gctccaattc ttggagtggt gaatccgtta
     4501 gcgaggtgcc gccggcttcc attcaggtcg aggtggcccg gctccatgca ccgcgacgca
     4561 acgcggggag gcagacaagg tatagggcgg cgcctacaat ccatgccaac ccgttccatg
     4621 tgctcgccga ggcggcataa atcgccgtga cgatcagcgg tccaatgatc gaagttaggc
     4681 tggtaagagc cgcgagcgat ccttgaagct gtccctgatg gtcgtcatct acctgcctgg
     4741 acagcatggc ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga atcataatgg
     4801 ggaaggccat ccagcctcgc gtcgcgaacg ccagcaagac gtagcccagc gcgtcggccg
     4861 ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga
     4921 aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg
     4981 cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta
     5041 cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc
     5101 accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgatcg acgctctccc
     5161 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc
     5221 gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct
     5281 gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc
     5341 ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg
     5401 gccacgatgc gtccggcgta gaggatcg
//
