LOCUS       sequence-458338-        7082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Anthoceros agrestis chaperones: 
            Raf1,Raf2,RbcX2,BSD2,RbcX1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7082)
  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 7082)
  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/Raf2/RbcX2/BSD2/RbcX1
FEATURES             Location/Qualifiers
     source          1..7082
                     /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      2004..2051
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2100..2118
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2119..2143
                     /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             2158..2180
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      2676..2723
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2768..2786
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2787..2811
                     /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             2826..2848
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      3164..3211
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3260..3278
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3279..3303
                     /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             3318..3340
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      3763..3810
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3981..4772)
                     /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(4773..4864)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4912..5650)
                     /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(5860..6942)
                     /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     6904..6923
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6943..7020)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(7066..7082,1..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 gagcacgtga tcctcattcc gccgcacctc ctgggcctcg ccgaccgcgc cgtcacaccc
     1621 cttgctccag gcacgcagcc tctatccgct gacgcggcca aagctctact tcgcaaggta
     1681 gttgcctgga gacttatcga ccagggaggc cttcgcatcg agtgcgagtg gaagctcaag
     1741 agttttggtg ctggcattga gctgttcgaa cgtatagctg aggtggcgga ggccgccggt
     1801 caccaccctg accttcatct cgaacaaggc aacagagcca gagtctgctt gtcgtctcac
     1861 tccgtcggag gcttgacgga gaatgacttc attgtagctg ccaggataga tcgcgtgaag
     1921 acatcggatc tcatattcaa gaagaacaag ttctgggcct gatcgaccag gcattactag
     1981 agtcacactc cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
     2041 ggggtttttt gaggtcgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt
     2101 aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt
     2161 gtttaacttt aagaaggaga tatacatatg gtgccttcat gtggtcaaca aggatcacgg
     2221 agacctcttg ttgtatgtga gcttggtggg aattatgagg aaagttttgc agatgttgac
     2281 aagcaccttc ttgattattt cacgttcaaa gcggtgaaga ctgttttggc acagttgtca
     2341 gagatgaacc ctgagcagta ttcctggttt tataactttg ttgtgagtaa tagacctcaa
     2401 gatagcaaat actttgtgcg cctgttagtg aaggaaaggc aagagcttgg tgagagggtc
     2461 atggtaacta gactacatct tttcaacgca tgggtgaaga agtacaacca tgcgcagatg
     2521 cacaaggcca tctcagagca gaacctcgag attctgaggg aacgtctcat ccaaacagtg
     2581 aaattgacat cagacgggga tgacttcgac aggccacctt tgaggggcca gtaatcgacc
     2641 aggcattact agagtcacac tccgctgagc aataactagc ataacccctt ggggcctcta
     2701 aacgggtctt gaggggtttt ttggcttgcg tccggcgtag aggatcgaga tctcgatccc
     2761 gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa
     2821 ataattttgt ttaactttaa gaaggagata tacatatggc ctctgcgcag ggggatgagt
     2881 ctaaggcgac ctcgcagtcg ggagtgaaga caaagagcat tctttgcatg aactgcacgg
     2941 ggaacggggc tgttgtgtgc tcccagtgta agggaaaagg cttgaattcg gtggatcact
     3001 tcaacggccg gtacaaagca ggccaaattt gctggctgtg caggggcaag aaacagatgc
     3061 tctgcggaga gtgcaacggt gccggattca ttggtggctt tctgaatacg caagacgatt
     3121 agtcgaccag gcattactag agtcacactc cgctgagcaa taactagcat aaccccttgg
     3181 ggcctctaaa cgggtcttga ggggtttttt gaggtcgatg cgtccggcgt agaggatcga
     3241 gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat
     3301 tcccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg tttgtgccag
     3361 gttttgggga gaagtctcca gagcggaagg cggcagattc cctgcacaac ttcttcacct
     3421 ttgtggctgt caagattgtt atgtcgcagc tccaggacta caacaaggaa ggatatgaag
     3481 agctgagcag ctttgtggac cgggtgtccc tcaaagatgg cgaccagttc tgtgccgcac
     3541 tattgcggga gtctcctcgt cacaaaggcc ttgctctccg catcatggag gttcgatcag
     3601 cgtacgctcg tcaggatttc gagtgggaca acttgcaagc gctggccatg aagcaagtgg
     3661 acgaggccaa caccagcctg atgcggaaat tcgtagtaga gactactacc ttccaggacc
     3721 agtgattaat taacctaggc tgctgccacc gctgagcaat aactagcata accccttggg
     3781 gcctctaaac gggtcttgag gggttttttg ctgaaacctc aggcatttga gaagcacacg
     3841 gtcacactgc ttccggtagt caataaaccg gtaaaccagc aatagacata agcggctatt
     3901 taacgaccct gccctgaacc gacgaccggg tcatcgtggc cggatcttgc ggcccctcgg
     3961 cttgaacgaa ttgttagaca ttatttgccg actaccttgg tgatctcgcc tttcacgtag
     4021 tggacaaatt cttccaactg atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga
     4081 taagcctgtc tagcttcaag tatgacgggc tgatactggg ccggcaggcg ctccattgcc
     4141 cagtcggcag cgacatcctt cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg
     4201 gacaacgtaa gcactacatt tcgctcatcg ccagcccagt cgggcggcga gttccatagc
     4261 gttaaggttt catttagcgc ctcaaataga tcctgttcag gaaccggatc aaagagttcc
     4321 tccgccgctg gacctaccaa ggcaacgcta tgttctcttg cttttgtcag caagatagcc
     4381 agatcaatgt cgatcgtggc tggctcgaag atacctgcaa gaatgtcatt gcgctgccat
     4441 tctccaaatt gcagttcgcg cttagctgga taacgccacg gaatgatgtc gtcgtgcaca
     4501 acaatggtga cttctacagc gcggagaatc tcgctctctc caggggaagc cgaagtttcc
     4561 aaaaggtcgt tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc
     4621 agcaaatcaa tatcactgtg tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt
     4681 acggccagca acgtcggttc gagatggcgc tcgatgacgc caactacctc tgatagttga
     4741 gtcgatactt cggcgatcac cgcttccctc atactcttcc tttttcaata ttattgaagc
     4801 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4861 caaatagcta gctcactcgg tcgctacgct ccgggcgtga gactgcggcg ggcgctgcgg
     4921 acacatacaa agttacccac agattccgtg gataagcagg ggactaacat gtgaggcaaa
     4981 acagcagggc cgcgccggtg gcgtttttcc ataggctccg ccctcctgcc agagttcaca
     5041 taaacagacg cttttccggt gcatctgtgg gagccgtgag gctcaaccat gaatctgaca
     5101 gtacgggcga aacccgacag gacttaaaga tccccaccgt ttccggcggg tcgctccctc
     5161 ttgcgctctc ctgttccgac cctgccgttt accggatacc tgttccgcct ttctccctta
     5221 cgggaagtgt ggcgctttct catagctcac acactggtat ctcggctcgg tgtaggtcgt
     5281 tcgctccaag ctgggctgta agcaagaact ccccgttcag cccgactgct gcgccttatc
     5341 cggtaactgt tcacttgagt ccaacccgga aaagcacggt aaaacgccac tggcagcagc
     5401 cattggtaac tgggagttcg cagaggattt gtttagctaa acacgcggtt gctcttgaag
     5461 tgtgcgccaa agtccggcta cactggaagg acagatttgg ttgctgtgct ctgcgaaagc
     5521 cagttaccac ggttaagcag ttccccaact gacttaacct tcgatcaaac cacctcccca
     5581 ggtggttttt tcgtttacag ggcaaaagat tacgcgcaga aaaaaaggat ctcaagaaga
     5641 tcctttgatc ttttctactg aaccgctcta gatttcagtg caatttatct cttcaaatgt
     5701 agcacctgaa gtcagcccca tacgatataa gttgtaattc tcatgttagt catgccccgc
     5761 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt
     5821 gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5881 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5941 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
     6001 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
     6061 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc
     6121 gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg
     6181 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
     6241 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
     6301 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
     6361 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
     6421 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     6481 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     6541 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
     6601 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     6661 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     6721 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     6781 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     6841 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     6901 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
     6961 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc
     7021 cgggatctcg acgctctccc ttatgcgact cctgcattag gaaattaata cgactcacta
     7081 ta
//
