LOCUS       sequence-460670-        7092 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Anthoceros agrestis chaperones: Raf1,RbcX2,BSD2,RbcX1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7092)
  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 7092)
  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_AaRaf1/RbcX2/BSD2/RbcX1
FEATURES             Location/Qualifiers
     source          1..7092
                     /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)."
     terminator      1288..1335
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1362..1380
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1381..1405
                     /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             1420..1442
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      2010..2057
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2106..2124
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2125..2149
                     /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             2164..2186
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      2682..2729
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2778..2796
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2797..2821
                     /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             2836..2858
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      3174..3221
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3270..3288
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3289..3313
                     /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             3328..3350
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      3773..3820
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3991..4782)
                     /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(4783..4874)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4922..5660)
                     /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(5870..6952)
                     /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     6914..6933
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6953..7030)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(7076..7092,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atggggggga gggacgttta ccagcctttc cggcccccgg tcaccaacct
      121 gcctgctccg ggtaccgtgt ccccggagga gcagctgcag atcctgaggg agaggcgggg
      181 gctatggcac gagtacgcgc tggtcatccc cttcctcacc cgagcaggct acactccggc
      241 gcagatcgat gaggcaaccg gggtgaccgg ggtggagcag aacatgatcg tgggggcgtc
      301 ccaggtgcgg ctgtcgctca tagccggggg aatgggagcg gaggtgctga atttcttcga
      361 tgtgggaggg gcagacctgc tgtacgagct cagaattctg tccacggatc agaggaaggc
      421 ttcggcggag ttgatcgtgg agaaaaagat ggaccctaaa gaggccaagg agctggctcg
      481 ggctgtgaag gactacgata ggaggaagag ggaggagggg tggggatcct tcaacaacgg
      541 caggccgggg gactatcttg ccttcgcttg ttaccgccag agcaaggagg tgagattcga
      601 tgctgagaaa gagattctga tcaacaaggg gctcagctat gccgcctctg actctgcacg
      661 gaccaagttg aaggaagccc tagggttgga agcagctccg gtgtcacccg agcgggagga
      721 ggccgttagg gaggaggccg ttaggtctag gttacaggtg gtgaggcttc ctagtaagca
      781 ggacacggaa aactctcccc tgcctgtcat actgcctgta gctactgacc tggcgactgt
      841 gaaaaaagct cctgcggttg gagcttttgg tgatggtcct tttaacgttt acggtgcaca
      901 gcctagatgg aagacatggc tagctcttcc aggatggtcc tctgtcgtga cgtcccgggc
      961 tcccattgct gtggtgttcc ctgatgcaac tatcttagat gtggggtcaa ctccgggaac
     1021 tgttgcaagg cagggcagaa acgagcctat tttgttggtg gtggacaagg gaaacgtcga
     1081 tccggaggaa gatgcttact ttctggtgga ggaaaaccag cgcttggctt tgcggttagg
     1141 ttcgaaacca gctgccacaa ccccaaagaa ctcaaagaca ttcgggaaag ttgtagttgt
     1201 attgagaccc cccctttctg aggacgagga tgcagtttgg gaatgatcga ccaggcatta
     1261 ctagagtcac actccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     1321 ttgaggggtt ttttgaatgc gagatctcga tcccgcgaaa ttaatacgac tcactatagg
     1381 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
     1441 gatatacata tggcacaggc cgtgtcgtcg gatcctttgg gagaggttgg agccagggat
     1501 ccaacatcct ctgaaatcga gagcagattc ggagataagg tgttgggaaa tgccagcacg
     1561 gagcactaat aagtgatcct cattccgccg cacctcctgg gcctcgccga ccgcgccgtc
     1621 acaccccttg ctccaggcac gcagcctcta tccgctgacg cggccaaagc tctacttcgc
     1681 aaggtagttg cctggagact tatcgaccag ggaggccttc gcatcgagtg cgagtggaag
     1741 ctcaagagtt ttggtgctgg cattgagctg ttcgaacgta tagctgaggt ggcggaggcc
     1801 gccggtcacc accctgacct tcatctcgaa caaggcaaca gagccagagt ctgcttgtcg
     1861 tctcactccg tcggaggctt gacggagaat gacttcattg tagctgccag gatagatcgc
     1921 gtgaagacat cggatctcat attcaagaag aacaagttct gggcctgatc gaccaggcat
     1981 tactagagtc acactccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     2041 tcttgagggg ttttttgagg tcgatgcgtc cggcgtagag gatcgagatc tcgatcccgc
     2101 gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc ctctagaaat
     2161 aattttgttt aactttaaga aggagatata catatggtgc cttcatgtgg tcaacaagga
     2221 tcacggagac ctcttgttgt atgtgagctt ggtgggaatt atgaggaaag ttttgcagat
     2281 gttgacaagc accttcttga ttatttcacg ttcaaagcgg tgaagactgt tttggcacag
     2341 ttgtcagaga tgaaccctga gcagtattcc tggttttata actttgttgt gagtaataga
     2401 cctcaagata gcaaatactt tgtgcgcctg ttagtgaagg aaaggcaaga gcttggtgag
     2461 agggtcatgg taactagact acatcttttc aacgcatggg tgaagaagta caaccatgcg
     2521 cagatgcaca aggccatctc agagcagaac ctcgagattc tgagggaacg tctcatccaa
     2581 acagtgaaat tgacatcaga cggggatgac ttcgacaggc cacctttgag gggccagtaa
     2641 tcgaccaggc attactagag tcacactccg ctgagcaata actagcataa ccccttgggg
     2701 cctctaaacg ggtcttgagg ggttttttga ggtcgatgcg tccggcgtag aggatcgaga
     2761 tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc
     2821 ccctctagaa ataattttgt ttaactttaa gaaggagata tacatatggc ctctgcgcag
     2881 ggggatgagt ctaaggcgac ctcgcagtcg ggagtgaaga caaagagcat tctttgcatg
     2941 aactgcacgg ggaacggggc tgttgtgtgc tcccagtgta agggaaaagg cttgaattcg
     3001 gtggatcact tcaacggccg gtacaaagca ggccaaattt gctggctgtg caggggcaag
     3061 aaacagatgc tctgcggaga gtgcaacggt gccggattca ttggtggctt tctgaatacg
     3121 caagacgatt agtcgaccag gcattactag agtcacactc cgctgagcaa taactagcat
     3181 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gaggtcgatg cgtccggcgt
     3241 agaggatcga gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc
     3301 ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg
     3361 tttgtgccag gttttgggga gaagtctcca gagcggaagg cggcagattc cctgcacaac
     3421 ttcttcacct ttgtggctgt caagattgtt atgtcgcagc tccaggacta caacaaggaa
     3481 ggatatgaag agctgagcag ctttgtggac cgggtgtccc tcaaagatgg cgaccagttc
     3541 tgtgccgcac tattgcggga gtctcctcgt cacaaaggcc ttgctctccg catcatggag
     3601 gttcgatcag cgtacgctcg tcaggatttc gagtgggaca acttgcaagc gctggccatg
     3661 aagcaagtgg acgaggccaa caccagcctg atgcggaaat tcgtagtaga gactactacc
     3721 ttccaggacc agtgattaat taacctaggc tgctgccacc gctgagcaat aactagcata
     3781 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaacctc aggcatttga
     3841 gaagcacacg gtcacactgc ttccggtagt caataaaccg gtaaaccagc aatagacata
     3901 agcggctatt taacgaccct gccctgaacc gacgaccggg tcatcgtggc cggatcttgc
     3961 ggcccctcgg cttgaacgaa ttgttagaca ttatttgccg actaccttgg tgatctcgcc
     4021 tttcacgtag tggacaaatt cttccaactg atctgcgcgc gaggccaagc gatcttcttc
     4081 ttgtccaaga taagcctgtc tagcttcaag tatgacgggc tgatactggg ccggcaggcg
     4141 ctccattgcc cagtcggcag cgacatcctt cggcgcgatt ttgccggtta ctgcgctgta
     4201 ccaaatgcgg gacaacgtaa gcactacatt tcgctcatcg ccagcccagt cgggcggcga
     4261 gttccatagc gttaaggttt catttagcgc ctcaaataga tcctgttcag gaaccggatc
     4321 aaagagttcc tccgccgctg gacctaccaa ggcaacgcta tgttctcttg cttttgtcag
     4381 caagatagcc agatcaatgt cgatcgtggc tggctcgaag atacctgcaa gaatgtcatt
     4441 gcgctgccat tctccaaatt gcagttcgcg cttagctgga taacgccacg gaatgatgtc
     4501 gtcgtgcaca acaatggtga cttctacagc gcggagaatc tcgctctctc caggggaagc
     4561 cgaagtttcc aaaaggtcgt tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt
     4621 caccgtaacc agcaaatcaa tatcactgtg tggcttcagg ccgccatcca ctgcggagcc
     4681 gtacaaatgt acggccagca acgtcggttc gagatggcgc tcgatgacgc caactacctc
     4741 tgatagttga gtcgatactt cggcgatcac cgcttccctc atactcttcc tttttcaata
     4801 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4861 gaaaaataaa caaatagcta gctcactcgg tcgctacgct ccgggcgtga gactgcggcg
     4921 ggcgctgcgg acacatacaa agttacccac agattccgtg gataagcagg ggactaacat
     4981 gtgaggcaaa acagcagggc cgcgccggtg gcgtttttcc ataggctccg ccctcctgcc
     5041 agagttcaca taaacagacg cttttccggt gcatctgtgg gagccgtgag gctcaaccat
     5101 gaatctgaca gtacgggcga aacccgacag gacttaaaga tccccaccgt ttccggcggg
     5161 tcgctccctc ttgcgctctc ctgttccgac cctgccgttt accggatacc tgttccgcct
     5221 ttctccctta cgggaagtgt ggcgctttct catagctcac acactggtat ctcggctcgg
     5281 tgtaggtcgt tcgctccaag ctgggctgta agcaagaact ccccgttcag cccgactgct
     5341 gcgccttatc cggtaactgt tcacttgagt ccaacccgga aaagcacggt aaaacgccac
     5401 tggcagcagc cattggtaac tgggagttcg cagaggattt gtttagctaa acacgcggtt
     5461 gctcttgaag tgtgcgccaa agtccggcta cactggaagg acagatttgg ttgctgtgct
     5521 ctgcgaaagc cagttaccac ggttaagcag ttccccaact gacttaacct tcgatcaaac
     5581 cacctcccca ggtggttttt tcgtttacag ggcaaaagat tacgcgcaga aaaaaaggat
     5641 ctcaagaaga tcctttgatc ttttctactg aaccgctcta gatttcagtg caatttatct
     5701 cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgtaattc tcatgttagt
     5761 catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg
     5821 agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc
     5881 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5941 aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca
     6001 gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt
     6061 gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt
     6121 cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc ccggactcgg
     6181 taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa
     6241 cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc
     6301 cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca
     6361 gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac
     6421 ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac
     6481 tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag
     6541 cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc
     6601 gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca
     6661 tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt
     6721 gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc
     6781 ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca
     6841 ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct
     6901 gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca
     6961 ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt
     7021 cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag gaaattaata
     7081 cgactcacta ta
//
