LOCUS       sequence-458336-        6625 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Anthoceros agrestis chaperones: Raf1,Raf2,RbcX2,RbcX1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6625)
  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 6625)
  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/RbcX1
FEATURES             Location/Qualifiers
     source          1..6625
                     /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      2707..2754
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2803..2821
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2822..2846
                     /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             2861..2883
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      3306..3353
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3524..4315)
                     /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(4316..4407)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4455..5193)
                     /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(5403..6485)
                     /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     6447..6466
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6486..6563)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(6609..6625,1..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 tccggtcgac caggcattac tagagtcaca ctccgctgag
     2701 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgaggtcg
     2761 atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga ctcactatag
     2821 gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg
     2881 agatatacat atgtttgtgc caggttttgg ggagaagtct ccagagcgga aggcggcaga
     2941 ttccctgcac aacttcttca cctttgtggc tgtcaagatt gttatgtcgc agctccagga
     3001 ctacaacaag gaaggatatg aagagctgag cagctttgtg gaccgggtgt ccctcaaaga
     3061 tggcgaccag ttctgtgccg cactattgcg ggagtctcct cgtcacaaag gccttgctct
     3121 ccgcatcatg gaggttcgat cagcgtacgc tcgtcaggat ttcgagtggg acaacttgca
     3181 agcgctggcc atgaagcaag tggacgaggc caacaccagc ctgatgcgga aattcgtagt
     3241 agagactact accttccagg accagtgatt aattaaccta ggctgctgcc accgctgagc
     3301 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaac
     3361 ctcaggcatt tgagaagcac acggtcacac tgcttccggt agtcaataaa ccggtaaacc
     3421 agcaatagac ataagcggct atttaacgac cctgccctga accgacgacc gggtcatcgt
     3481 ggccggatct tgcggcccct cggcttgaac gaattgttag acattatttg ccgactacct
     3541 tggtgatctc gcctttcacg tagtggacaa attcttccaa ctgatctgcg cgcgaggcca
     3601 agcgatcttc ttcttgtcca agataagcct gtctagcttc aagtatgacg ggctgatact
     3661 gggccggcag gcgctccatt gcccagtcgg cagcgacatc cttcggcgcg attttgccgg
     3721 ttactgcgct gtaccaaatg cgggacaacg taagcactac atttcgctca tcgccagccc
     3781 agtcgggcgg cgagttccat agcgttaagg tttcatttag cgcctcaaat agatcctgtt
     3841 caggaaccgg atcaaagagt tcctccgccg ctggacctac caaggcaacg ctatgttctc
     3901 ttgcttttgt cagcaagata gccagatcaa tgtcgatcgt ggctggctcg aagatacctg
     3961 caagaatgtc attgcgctgc cattctccaa attgcagttc gcgcttagct ggataacgcc
     4021 acggaatgat gtcgtcgtgc acaacaatgg tgacttctac agcgcggaga atctcgctct
     4081 ctccagggga agccgaagtt tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat
     4141 caagccttac ggtcaccgta accagcaaat caatatcact gtgtggcttc aggccgccat
     4201 ccactgcgga gccgtacaaa tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga
     4261 cgccaactac ctctgatagt tgagtcgata cttcggcgat caccgcttcc ctcatactct
     4321 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4381 ttgaatgtat ttagaaaaat aaacaaatag ctagctcact cggtcgctac gctccgggcg
     4441 tgagactgcg gcgggcgctg cggacacata caaagttacc cacagattcc gtggataagc
     4501 aggggactaa catgtgaggc aaaacagcag ggccgcgccg gtggcgtttt tccataggct
     4561 ccgccctcct gccagagttc acataaacag acgcttttcc ggtgcatctg tgggagccgt
     4621 gaggctcaac catgaatctg acagtacggg cgaaacccga caggacttaa agatccccac
     4681 cgtttccggc gggtcgctcc ctcttgcgct ctcctgttcc gaccctgccg tttaccggat
     4741 acctgttccg cctttctccc ttacgggaag tgtggcgctt tctcatagct cacacactgg
     4801 tatctcggct cggtgtaggt cgttcgctcc aagctgggct gtaagcaaga actccccgtt
     4861 cagcccgact gctgcgcctt atccggtaac tgttcacttg agtccaaccc ggaaaagcac
     4921 ggtaaaacgc cactggcagc agccattggt aactgggagt tcgcagagga tttgtttagc
     4981 taaacacgcg gttgctcttg aagtgtgcgc caaagtccgg ctacactgga aggacagatt
     5041 tggttgctgt gctctgcgaa agccagttac cacggttaag cagttcccca actgacttaa
     5101 ccttcgatca aaccacctcc ccaggtggtt ttttcgttta cagggcaaaa gattacgcgc
     5161 agaaaaaaag gatctcaaga agatcctttg atcttttcta ctgaaccgct ctagatttca
     5221 gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc ccatacgata taagttgtaa
     5281 ttctcatgtt agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     5341 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc
     5401 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5461 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt
     5521 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg
     5581 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata
     5641 taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatgtccgc accaacgcgc
     5701 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc
     5761 atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg
     5821 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta
     5881 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg
     5941 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg
     6001 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca
     6061 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc
     6121 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg
     6181 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc
     6241 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc
     6301 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc
     6361 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg
     6421 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
     6481 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag
     6541 gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat
     6601 taggaaatta atacgactca ctata
//
