LOCUS       sequence-460708-        6083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Anthoceros agrestis chaperones: Raf2,RbcX2,BSD2,RbcX1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6083)
  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 6083)
  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_AaRaf2/RbcX2/BSD2/RbcX1
FEATURES             Location/Qualifiers
     source          1..6083
                     /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)."
     terminator      302..349
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        376..394
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    395..419
                     /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             434..456
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      1018..1065
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1114..1132
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1133..1157
                     /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             1172..1194
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      1690..1737
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1786..1804
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1805..1829
                     /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             1844..1866
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      2182..2229
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2278..2296
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2297..2321
                     /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             2336..2358
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      2781..2828
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2999..3790)
                     /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(3791..3882)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3930..4668)
                     /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(4878..5960)
                     /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     5922..5941
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5961..6038)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taataaggag atataccatg gtgatccgga ggaagatgct tactttctgg
      121 tggaggaaaa ccagcgcttg gctttgcggt taggttcgaa accagctgcc acaaccccaa
      181 agaactcaaa gacattcggg aaagttgtag ttgtattgag accccccctt tctgaggacg
      241 aggatgcagt ttgggaatga tcgaccaggc attactagag tcacactccg ctgagcaata
      301 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttga atgcgagatc
      361 tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc
      421 ctctagaaat aattttgttt aactttaaga aggagatata catatggcac aggccgtgtc
      481 gtcggatcct ttgggagagg ttggagccag ggatccaaca tcctctgaaa tcgagagcag
      541 attcggagat aaggtgttgg gaaatgccag cacggagcac gtgatcctca ttccgccgca
      601 cctcctgggc ctcgccgacc gcgccgtcac accccttgct ccaggcacgc agcctctatc
      661 cgctgacgcg gccaaagctc tacttcgcaa ggtagttgcc tggagactta tcgaccaggg
      721 aggccttcgc atcgagtgcg agtggaagct caagagtttt ggtgctggca ttgagctgtt
      781 cgaacgtata gctgaggtgg cggaggccgc cggtcaccac cctgaccttc atctcgaaca
      841 aggcaacaga gccagagtct gcttgtcgtc tcactccgtc ggaggcttga cggagaatga
      901 cttcattgta gctgccagga tagatcgcgt gaagacatcg gatctcatat tcaagaagaa
      961 caagttctgg gcctgatcga ccaggcatta ctagagtcac actccgctga gcaataacta
     1021 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgaggtc gatgcgtccg
     1081 gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt
     1141 gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatataca
     1201 tatggtgcct tcatgtggtc aacaaggatc acggagacct cttgttgtat gtgagcttgg
     1261 tgggaattat gaggaaagtt ttgcagatgt tgacaagcac cttcttgatt atttcacgtt
     1321 caaagcggtg aagactgttt tggcacagtt gtcagagatg aaccctgagc agtattcctg
     1381 gttttataac tttgttgtga gtaatagacc tcaagatagc aaatactttg tgcgcctgtt
     1441 agtgaaggaa aggcaagagc ttggtgagag ggtcatggta actagactac atcttttcaa
     1501 cgcatgggtg aagaagtaca accatgcgca gatgcacaag gccatctcag agcagaacct
     1561 cgagattctg agggaacgtc tcatccaaac agtgaaattg acatcagacg gggatgactt
     1621 cgacaggcca cctttgaggg gccagtaatc gaccaggcat tactagagtc acactccgct
     1681 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgagg
     1741 tcgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta
     1801 taggggaatt gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga
     1861 aggagatata catatggcct ctgcgcaggg ggatgagtct aaggcgacct cgcagtcggg
     1921 agtgaagaca aagagcattc tttgcatgaa ctgcacgggg aacggggctg ttgtgtgctc
     1981 ccagtgtaag ggaaaaggct tgaattcggt ggatcacttc aacggccggt acaaagcagg
     2041 ccaaatttgc tggctgtgca ggggcaagaa acagatgctc tgcggagagt gcaacggtgc
     2101 cggattcatt ggtggctttc tgaatacgca agacgattag tcgaccaggc attactagag
     2161 tcacactccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     2221 ggttttttga ggtcgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaattaa
     2281 tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa ataattttgt
     2341 ttaactttaa gaaggagata tacatatgtt tgtgccaggt tttggggaga agtctccaga
     2401 gcggaaggcg gcagattccc tgcacaactt cttcaccttt gtggctgtca agattgttat
     2461 gtcgcagctc caggactaca acaaggaagg atatgaagag ctgagcagct ttgtggaccg
     2521 ggtgtccctc aaagatggcg accagttctg tgccgcacta ttgcgggagt ctcctcgtca
     2581 caaaggcctt gctctccgca tcatggaggt tcgatcagcg tacgctcgtc aggatttcga
     2641 gtgggacaac ttgcaagcgc tggccatgaa gcaagtggac gaggccaaca ccagcctgat
     2701 gcggaaattc gtagtagaga ctactacctt ccaggaccag tgattaatta acctaggctg
     2761 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     2821 gttttttgct gaaacctcag gcatttgaga agcacacggt cacactgctt ccggtagtca
     2881 ataaaccggt aaaccagcaa tagacataag cggctattta acgaccctgc cctgaaccga
     2941 cgaccgggtc atcgtggccg gatcttgcgg cccctcggct tgaacgaatt gttagacatt
     3001 atttgccgac taccttggtg atctcgcctt tcacgtagtg gacaaattct tccaactgat
     3061 ctgcgcgcga ggccaagcga tcttcttctt gtccaagata agcctgtcta gcttcaagta
     3121 tgacgggctg atactgggcc ggcaggcgct ccattgccca gtcggcagcg acatccttcg
     3181 gcgcgatttt gccggttact gcgctgtacc aaatgcggga caacgtaagc actacatttc
     3241 gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt taaggtttca tttagcgcct
     3301 caaatagatc ctgttcagga accggatcaa agagttcctc cgccgctgga cctaccaagg
     3361 caacgctatg ttctcttgct tttgtcagca agatagccag atcaatgtcg atcgtggctg
     3421 gctcgaagat acctgcaaga atgtcattgc gctgccattc tccaaattgc agttcgcgct
     3481 tagctggata acgccacgga atgatgtcgt cgtgcacaac aatggtgact tctacagcgc
     3541 ggagaatctc gctctctcca ggggaagccg aagtttccaa aaggtcgttg atcaaagctc
     3601 gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag caaatcaata tcactgtgtg
     3661 gcttcaggcc gccatccact gcggagccgt acaaatgtac ggccagcaac gtcggttcga
     3721 gatggcgctc gatgacgcca actacctctg atagttgagt cgatacttcg gcgatcaccg
     3781 cttccctcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     3841 tgagcggata catatttgaa tgtatttaga aaaataaaca aatagctagc tcactcggtc
     3901 gctacgctcc gggcgtgaga ctgcggcggg cgctgcggac acatacaaag ttacccacag
     3961 attccgtgga taagcagggg actaacatgt gaggcaaaac agcagggccg cgccggtggc
     4021 gtttttccat aggctccgcc ctcctgccag agttcacata aacagacgct tttccggtgc
     4081 atctgtggga gccgtgaggc tcaaccatga atctgacagt acgggcgaaa cccgacagga
     4141 cttaaagatc cccaccgttt ccggcgggtc gctccctctt gcgctctcct gttccgaccc
     4201 tgccgtttac cggatacctg ttccgccttt ctcccttacg ggaagtgtgg cgctttctca
     4261 tagctcacac actggtatct cggctcggtg taggtcgttc gctccaagct gggctgtaag
     4321 caagaactcc ccgttcagcc cgactgctgc gccttatccg gtaactgttc acttgagtcc
     4381 aacccggaaa agcacggtaa aacgccactg gcagcagcca ttggtaactg ggagttcgca
     4441 gaggatttgt ttagctaaac acgcggttgc tcttgaagtg tgcgccaaag tccggctaca
     4501 ctggaaggac agatttggtt gctgtgctct gcgaaagcca gttaccacgg ttaagcagtt
     4561 ccccaactga cttaaccttc gatcaaacca cctccccagg tggttttttc gtttacaggg
     4621 caaaagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctactgaa
     4681 ccgctctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt cagccccata
     4741 cgatataagt tgtaattctc atgttagtca tgccccgcgc ccaccggaag gagctgactg
     4801 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
     4861 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4921 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     4981 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     5041 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     5101 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg
     5161 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     5221 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     5281 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     5341 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     5401 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     5461 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     5521 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     5581 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     5641 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     5701 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     5761 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     5821 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     5881 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     5941 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     6001 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
     6061 atgcgactcc tgcattagga aat
//
