LOCUS       sequence-460710-        7587 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Arabidopsis thaliana chaperones: 
            Raf1,Raf2,RbcX2,BSD2,RbcX1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7587)
  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 7587)
  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/RbcX2/BSD2/RbcX1
FEATURES             Location/Qualifiers
     source          1..7587
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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        1386..1404
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1405..1429
                     /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             1444..1466
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1993..2022
                     /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      2090..2137
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2304..2322
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2323..2347
                     /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             2362..2384
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2821..2850
                     /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      2918..2965
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3132..3150
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3151..3175
                     /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             3190..3212
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3469..3498
                     /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      3566..3613
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3773..3791
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3792..3816
                     /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             3831..3853
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      4285..4332
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(4503..5294)
                     /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(5295..5386)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5434..6172)
                     /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(6382..7464)
                     /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     7426..7445
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7465..7542)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taataaggag atataccatg ggtatgcaac agctctacca accgttccga
      121 ccaccgtcat ctcccattcc aacccaattc cgttcactcg actccgccgg taaaatcgaa
      181 atcctcgccg gtcgaatggc tctctggttc gaatacgcac ctcttatctc ttctctttac
      241 accgatggtt tcactcctcc aaccatcgaa gaactcaccg gaatctcaag catcgaacag
      301 aaccgtttaa tcgtcggtgc gcaagttcgt gactcaattc ttcaatctat ccatgaaccg
      361 gaacttattt ctgcgttcga caccggtggt gcagagcttc tatacgaaat tcgtctcctt
      421 agcaccacgc agcgtgttgc agccgccaca ttcatcattg atcgtaatat tgattcgaaa
      481 ggagcacagg atttggcacg agccatcaag gattatccta accgtcgtgg agacgttgga
      541 tggttggatt ttgattacaa cttaccagga gattgtcttt cattcttgta ttacaggcag
      601 agcagagaga ataagaatcc atcggatcag agaacctcga tgcttctaca agcattgggg
      661 gttgcagaat cagaaaaggc taagaacagg ttgaacacag agctatacgg ggacaaggaa
      721 gcagagaagg agaaggagaa gaagaagaag gaagaggaag tcaaagctat tcggattcca
      781 gtggtgaggc ttaagtttgg agaagtagca gaagcgacct cggtggtagt tttaccggtt
      841 tgtaaagcag aggaagggga aaagaagatt cttgaagctc caatggagat catagccgga
      901 ggggatttta aggtggttga ggcggagaaa ggatggaaga gatgggtagt gcttccgtcg
      961 tggaacccag tggcagccat tgggaaaggc ggtgtggcgg tttctttcag ggatgataga
     1021 aaagtgctgc cttgggatgg aaaggaggag cctttactgg tagtggccga tagggtgagg
     1081 aatgttgtgg aagctgatga cgggtattat ctcgtggtgg ctgagaacgg acttaagcta
     1141 gagaaaggat cagatttgaa ggcgagagag gtgaaggaga gtttagggat ggttgttttg
     1201 gtggtgaggc cgccaagaga agatgatgat gattggcaga caagtcatca gaactgggac
     1261 tgacaattcg agctcggcgc gcctgcaggt cggcgatata ggcgccagca accgcacctg
     1321 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc
     1381 gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc ctctagaaat
     1441 aattttgttt aactttaaga aggagatata catatgatgt ctaatctggc gcaggatttt
     1501 cttggtgact tcggagctcg tgacccttac ccggaggaga tagcgagtca attcggagat
     1561 aaagtgttgg gatgccaaag cactgagcac aagattttga taccaaacgc atctgttttg
     1621 tctctctccc agcttcagtg ttcccctgtt tcgtcttcac agcctccttt gtccggcgat
     1681 gatgccagaa ctctcctcca caaggttttg ggatggagta tagtggataa tgaagcgggt
     1741 ggtctgaaaa taaggtgtat gtggaaagtg agggattttg ggtgcggtgt tgaactcata
     1801 aacaggatcc ataaggttgc tgaagcttct ggtcattacc cttctcttca cttggaaagt
     1861 cctacccaag ttcgagctga actatttacc tcttctatcg gagggttgag catgaatgat
     1921 ttcataatgg cggctaaaat agatgatatc aagacttctg atctttcccc taggaaaaga
     1981 gcttgggcgt gagctagcat gactggtgga cagcaaatgg gtcgcggatc cggctgctaa
     2041 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     2101 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     2161 atatcccgca agaggcccgg cagtaccggc ataaccaagc gtcgacgtcg gcgatatagg
     2221 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
     2281 tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
     2341 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatataca tatggtatcg
     2401 agaattgcga atcgaaagag caaaaagctc ttgattgtga atgaggatgt tgctggtaat
     2461 tacgacgata ccttcggtga tgtacaaaag caaattgtta attattttac atacaaagct
     2521 gtgaggacgg ttcttcatca gctgtacgaa atgaatcctc ctcaatacac ttggttctat
     2581 aatcatatca taacaaacag accgacggat ggcaaacgtt tcctccgtgc cctcggcaag
     2641 gagagtcagg agcttgcaga aagagtgatg atcacgcgtc ttcacttgta tggcaaatgg
     2701 atcaagaaat gcgaccatgg gaagatatac caggaaatat cggatgaaaa cttggcgttg
     2761 atgcgtgaac gcctgatgga gaccgtgata tggccttccg atgacacaaa ctcaaggtaa
     2821 gctagcatga ctggtggaca gcaaatgggt cgcggatccg gctgctaaca aagcccgaaa
     2881 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     2941 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat atcccgcaag
     3001 aggcccggca gtaccggcat aaccaagcgt cgacgtcggc gatataggcg ccagcaaccg
     3061 cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatctcga
     3121 tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct
     3181 agaaataatt ttgtttaact ttaagaagga gatatacata tgatggccgc aaacaataat
     3241 cctcagggca ctaaaccaaa cagtttagtt tgtgcaaatt gtgaagggga aggttgtgtg
     3301 gcatgttctc aatgcaaagg aggtggtgtg aatttgattg atcatttcaa tggtcagttt
     3361 aaagctggtg cactctgttg gctttgcaga ggtaaaaagg aggtgttgtg tggggactgc
     3421 aatggagctg gtttcattgg tgggttcttg agcaccttcg atgagtaggc tagcatgact
     3481 ggtggacagc aaatgggtcg cggatccggc tgctaacaaa gcccgaaagg aagctgagtt
     3541 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     3601 gaggggtttt ttgctgaaag gaggaactat atccggatat cccgcaagag gcccggcagt
     3661 accggcataa ccaagcggta ccagcggtca attgtagtgg aggtgctagc gcatgcggaa
     3721 ttggatatcg gccggccacg cgatcgctga cgtcggtacc ctcgagtctg gttaatacga
     3781 ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac
     3841 tttaagaagg agatatacat atgtatgttc ccggctttgg agaagcttca ccggaggcta
     3901 aggcagcgaa gcatcttcat gacttcttta cttacgttgc agtgaggata gtgtctgctc
     3961 agcttgagag ttataatcct gaggcttata tggagttgag agaattttta gatacaaact
     4021 ctgtaagtga cggtgataaa ttctgcgcca ctctcatgcg tcgctcttca cgtcacatga
     4081 acttagccct tcgaatttta gaggtacggt ctgcttattg taaaaacgat ttcgaatggg
     4141 ataatatgaa gcgcctcgcc ttcaagaacg tagatgattc caacacaaga ctcatgcgcg
     4201 agtacgtctt ggagactagc catgtcgaaa ccgattctga taagtgatta attaacctag
     4261 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     4321 aggggttttt tgctgaaacc tcaggcattt gagaagcaca cggtcacact gcttccggta
     4381 gtcaataaac cggtaaacca gcaatagaca taagcggcta tttaacgacc ctgccctgaa
     4441 ccgacgaccg ggtcatcgtg gccggatctt gcggcccctc ggcttgaacg aattgttaga
     4501 cattatttgc cgactacctt ggtgatctcg cctttcacgt agtggacaaa ttcttccaac
     4561 tgatctgcgc gcgaggccaa gcgatcttct tcttgtccaa gataagcctg tctagcttca
     4621 agtatgacgg gctgatactg ggccggcagg cgctccattg cccagtcggc agcgacatcc
     4681 ttcggcgcga ttttgccggt tactgcgctg taccaaatgc gggacaacgt aagcactaca
     4741 tttcgctcat cgccagccca gtcgggcggc gagttccata gcgttaaggt ttcatttagc
     4801 gcctcaaata gatcctgttc aggaaccgga tcaaagagtt cctccgccgc tggacctacc
     4861 aaggcaacgc tatgttctct tgcttttgtc agcaagatag ccagatcaat gtcgatcgtg
     4921 gctggctcga agatacctgc aagaatgtca ttgcgctgcc attctccaaa ttgcagttcg
     4981 cgcttagctg gataacgcca cggaatgatg tcgtcgtgca caacaatggt gacttctaca
     5041 gcgcggagaa tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc gttgatcaaa
     5101 gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc aatatcactg
     5161 tgtggcttca ggccgccatc cactgcggag ccgtacaaat gtacggccag caacgtcggt
     5221 tcgagatggc gctcgatgac gccaactacc tctgatagtt gagtcgatac ttcggcgatc
     5281 accgcttccc tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5341 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagc tagctcactc
     5401 ggtcgctacg ctccgggcgt gagactgcgg cgggcgctgc ggacacatac aaagttaccc
     5461 acagattccg tggataagca ggggactaac atgtgaggca aaacagcagg gccgcgccgg
     5521 tggcgttttt ccataggctc cgccctcctg ccagagttca cataaacaga cgcttttccg
     5581 gtgcatctgt gggagccgtg aggctcaacc atgaatctga cagtacgggc gaaacccgac
     5641 aggacttaaa gatccccacc gtttccggcg ggtcgctccc tcttgcgctc tcctgttccg
     5701 accctgccgt ttaccggata cctgttccgc ctttctccct tacgggaagt gtggcgcttt
     5761 ctcatagctc acacactggt atctcggctc ggtgtaggtc gttcgctcca agctgggctg
     5821 taagcaagaa ctccccgttc agcccgactg ctgcgcctta tccggtaact gttcacttga
     5881 gtccaacccg gaaaagcacg gtaaaacgcc actggcagca gccattggta actgggagtt
     5941 cgcagaggat ttgtttagct aaacacgcgg ttgctcttga agtgtgcgcc aaagtccggc
     6001 tacactggaa ggacagattt ggttgctgtg ctctgcgaaa gccagttacc acggttaagc
     6061 agttccccaa ctgacttaac cttcgatcaa accacctccc caggtggttt tttcgtttac
     6121 agggcaaaag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6181 tgaaccgctc tagatttcag tgcaatttat ctcttcaaat gtagcacctg aagtcagccc
     6241 catacgatat aagttgtaat tctcatgtta gtcatgcccc gcgcccaccg gaaggagctg
     6301 actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa
     6361 cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     6421 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt
     6481 ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg
     6541 agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat
     6601 ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga
     6661 gatgtccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat
     6721 ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt
     6781 ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg
     6841 attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa
     6901 tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag
     6961 tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc
     7021 aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc
     7081 cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc
     7141 tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg
     7201 atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga
     7261 ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg
     7321 aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg
     7381 gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac
     7441 atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta
     7501 tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc
     7561 ccttatgcga ctcctgcatt aggaaat
//
