LOCUS       sequence-458337-        6311 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Arabidopsis thaliana chaperones: Raf1,Raf2,BSD2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6311)
  AUTHORS   Oh ZG, Robison TA, Loh DH, Ang WSL, Ng JZY, Li FW, Gunn LH
  TITLE     Unique biogenesis and kinetics of hornwort Rubiscos revealed by 
            synthetic biology systems.
  JOURNAL   Mol Plant. 2024 Dec 2;17(12):1833-1849. doi: 
            10.1016/j.molp.2024.10.013. Epub 2024 Nov 2.
  PUBMED    39491367
REFERENCE   2  (bases 1 to 6311)
  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: 
            "10.1016/j.molp.2024.10.013"; journalName: "Mol Plant"; date: 
            "2024-12-2- 2"; volume: "17"; issue: "12"; pages: "1833-1849"
COMMENT     Sequence Label: pCDF_AtRaf1/Raf2/BSD2
FEATURES             Location/Qualifiers
     source          1..6311
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     promoter        1369..1387
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1388..1412
                     /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             1427..1449
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1976..2005
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="ASMTGGQQMG"
     terminator      2073..2120
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2293..2311
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2312..2336
                     /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             2351..2373
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2630..2659
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="ASMTGGQQMG"
     terminator      2727..2774
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2992..3039
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3210..4001)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(4002..4093)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4141..4879)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(5089..6171)
                     /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     6133..6152
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6172..6249)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(6295..6311,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggtatgc aacagctcta ccaaccgttc cgaccaccgt catctcccat
      121 tccaacccaa ttccgttcac tcgactccgc cggtaaaatc gaaatcctcg ccggtcgaat
      181 ggctctctgg ttcgaatacg cacctcttat ctcttctctt tacaccgatg gtttcactcc
      241 tccaaccatc gaagaactca ccggaatctc aagcatcgaa cagaaccgtt taatcgtcgg
      301 tgcgcaagtt cgtgactcaa ttcttcaatc tatccatgaa ccggaactta tttctgcgtt
      361 cgacaccggt ggtgcagagc ttctatacga aattcgtctc cttagcacca cgcagcgtgt
      421 tgcagccgcc acattcatca ttgatcgtaa tattgattcg aaaggagcac aggatttggc
      481 acgagccatc aaggattatc ctaaccgtcg tggagacgtt ggatggttgg attttgatta
      541 caacttacca ggagattgtc tttcattctt gtattacagg cagagcagag agaataagaa
      601 tccatcggat cagagaacct cgatgcttct acaagcattg ggggttgcag aatcagaaaa
      661 ggctaagaac aggttgaaca cagagctata cggggacaag gaagcagaga aggagaagga
      721 gaagaagaag aaggaagagg aagtcaaagc tattcggatt ccagtggtga ggcttaagtt
      781 tggagaagta gcagaagcga cctcggtggt agttttaccg gtttgtaaag cagaggaagg
      841 ggaaaagaag attcttgaag ctccaatgga gatcatagcc ggaggggatt ttaaggtggt
      901 tgaggcggag aaaggatgga agagatgggt agtgcttccg tcgtggaacc cagtggcagc
      961 cattgggaaa ggcggtgtgg cggtttcttt cagggatgat agaaaagtgc tgccttggga
     1021 tggaaaggag gagcctttac tggtagtggc cgatagggtg aggaatgttg tggaagctga
     1081 tgacgggtat tatctcgtgg tggctgagaa cggacttaag ctagagaaag gatcagattt
     1141 gaaggcgaga gaggtgaagg agagtttagg gatggttgtt ttggtggtga ggccgccaag
     1201 agaagatgat gatgattggc agacaagtca tcagaactgg gactgacaat tcgagctcgg
     1261 cgcgcctgca ggtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg
     1321 gccacgatgc gtccggcgta gaggatcgag atctcgatcc cgcgaaatta atacgactca
     1381 ctatagggga attgtgagcg gataacaatt cccctctaga aataattttg tttaacttta
     1441 agaaggagat atacatatga tgtctaatct ggcgcaggat tttcttggtg acttcggagc
     1501 tcgtgaccct tacccggagg agatagcgag tcaattcgga gataaagtgt tgggatgcca
     1561 aagcactgag cacaagattt tgataccaaa cgcatctgtt ttgtctctct cccagcttca
     1621 gtgttcccct gtttcgtctt cacagcctcc tttgtccggc gatgatgcca gaactctcct
     1681 ccacaaggtt ttgggatgga gtatagtgga taatgaagcg ggtggtctga aaataaggtg
     1741 tatgtggaaa gtgagggatt ttgggtgcgg tgttgaactc ataaacagga tccataaggt
     1801 tgctgaagct tctggtcatt acccttctct tcacttggaa agtcctaccc aagttcgagc
     1861 tgaactattt acctcttcta tcggagggtt gagcatgaat gatttcataa tggcggctaa
     1921 aatagatgat atcaagactt ctgatctttc ccctaggaaa agagcttggg cgtgagctag
     1981 catgactggt ggacagcaaa tgggtcgcgg atccggctgc taacaaagcc cgaaaggaag
     2041 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     2101 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatatccc gcaagaggcc
     2161 cggcagtacc ggcataacca agcgtcgaca ctagtgtcgg cgatataggc gccagcaacc
     2221 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg
     2281 atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac aattcccctc
     2341 tagaaataat tttgtttaac tttaagaagg agatatacat atgatggccg caaacaataa
     2401 tcctcagggc actaaaccaa acagtttagt ttgtgcaaat tgtgaagggg aaggttgtgt
     2461 ggcatgttct caatgcaaag gaggtggtgt gaatttgatt gatcatttca atggtcagtt
     2521 taaagctggt gcactctgtt ggctttgcag aggtaaaaag gaggtgttgt gtggggactg
     2581 caatggagct ggtttcattg gtgggttctt gagcaccttc gatgagtagg ctagcatgac
     2641 tggtggacag caaatgggtc gcggatccgg ctgctaacaa agcccgaaag gaagctgagt
     2701 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
     2761 tgaggggttt tttgctgaaa ggaggaacta tatccggata tcccgcaaga ggcccggcag
     2821 taccggcata accaagcggt accagcggtc aattgtagtg gaggtgctag cgcatgcgga
     2881 attggatatc ggccggccac gcgatcgctg acgtcggtac cctcgagtct ggtcctctag
     2941 aaataattat acatttaatt aacctaggct gctgccaccg ctgagcaata actagcataa
     3001 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaacctca ggcatttgag
     3061 aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa
     3121 gcggctattt aacgaccctg ccctgaaccg acgaccgggt catcgtggcc ggatcttgcg
     3181 gcccctcggc ttgaacgaat tgttagacat tatttgccga ctaccttggt gatctcgcct
     3241 ttcacgtagt ggacaaattc ttccaactga tctgcgcgcg aggccaagcg atcttcttct
     3301 tgtccaagat aagcctgtct agcttcaagt atgacgggct gatactgggc cggcaggcgc
     3361 tccattgccc agtcggcagc gacatccttc ggcgcgattt tgccggttac tgcgctgtac
     3421 caaatgcggg acaacgtaag cactacattt cgctcatcgc cagcccagtc gggcggcgag
     3481 ttccatagcg ttaaggtttc atttagcgcc tcaaatagat cctgttcagg aaccggatca
     3541 aagagttcct ccgccgctgg acctaccaag gcaacgctat gttctcttgc ttttgtcagc
     3601 aagatagcca gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag aatgtcattg
     3661 cgctgccatt ctccaaattg cagttcgcgc ttagctggat aacgccacgg aatgatgtcg
     3721 tcgtgcacaa caatggtgac ttctacagcg cggagaatct cgctctctcc aggggaagcc
     3781 gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag ccttacggtc
     3841 accgtaacca gcaaatcaat atcactgtgt ggcttcaggc cgccatccac tgcggagccg
     3901 tacaaatgta cggccagcaa cgtcggttcg agatggcgct cgatgacgcc aactacctct
     3961 gatagttgag tcgatacttc ggcgatcacc gcttccctca tactcttcct ttttcaatat
     4021 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4081 aaaaataaac aaatagctag ctcactcggt cgctacgctc cgggcgtgag actgcggcgg
     4141 gcgctgcgga cacatacaaa gttacccaca gattccgtgg ataagcaggg gactaacatg
     4201 tgaggcaaaa cagcagggcc gcgccggtgg cgtttttcca taggctccgc cctcctgcca
     4261 gagttcacat aaacagacgc ttttccggtg catctgtggg agccgtgagg ctcaaccatg
     4321 aatctgacag tacgggcgaa acccgacagg acttaaagat ccccaccgtt tccggcgggt
     4381 cgctccctct tgcgctctcc tgttccgacc ctgccgttta ccggatacct gttccgcctt
     4441 tctcccttac gggaagtgtg gcgctttctc atagctcaca cactggtatc tcggctcggt
     4501 gtaggtcgtt cgctccaagc tgggctgtaa gcaagaactc cccgttcagc ccgactgctg
     4561 cgccttatcc ggtaactgtt cacttgagtc caacccggaa aagcacggta aaacgccact
     4621 ggcagcagcc attggtaact gggagttcgc agaggatttg tttagctaaa cacgcggttg
     4681 ctcttgaagt gtgcgccaaa gtccggctac actggaagga cagatttggt tgctgtgctc
     4741 tgcgaaagcc agttaccacg gttaagcagt tccccaactg acttaacctt cgatcaaacc
     4801 acctccccag gtggtttttt cgtttacagg gcaaaagatt acgcgcagaa aaaaaggatc
     4861 tcaagaagat cctttgatct tttctactga accgctctag atttcagtgc aatttatctc
     4921 ttcaaatgta gcacctgaag tcagccccat acgatataag ttgtaattct catgttagtc
     4981 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
     5041 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     5101 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     5161 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     5221 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     5281 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     5341 ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt
     5401 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     5461 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     5521 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     5581 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     5641 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     5701 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     5761 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     5821 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     5881 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     5941 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     6001 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     6061 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     6121 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     6181 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     6241 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aaattaatac
     6301 gactcactat a
//
