LOCUS       sequence-468479-        5859 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Anthoceros agrestis chaperones: Raf1,Raf2,BSD2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5859)
  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 5859)
  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/BSD2
FEATURES             Location/Qualifiers
     source          1..5859
                     /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        2144..2162
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2163..2187
                     /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             2202..2224
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      2540..2587
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2758..3549)
                     /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(3550..3641)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3689..4427)
                     /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(4637..5719)
                     /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     5681..5700
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5720..5797)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(5843..5859,1..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 ccgcgaaatg
     2101 cttgcgtccg gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata
     2161 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
     2221 gagatataca tatggcctct gcgcaggggg atgagtctaa ggcgacctcg cagtcgggag
     2281 tgaagacaaa gagcattctt tgcatgaact gcacggggaa cggggctgtt gtgtgctccc
     2341 agtgtaaggg aaaaggcttg aattcggtgg atcacttcaa cggccggtac aaagcaggcc
     2401 aaatttgctg gctgtgcagg ggcaagaaac agatgctctg cggagagtgc aacggtgccg
     2461 gattcattgg tggctttctg aatacgcaag acgattagtc gaccaggcat tactagagtc
     2521 acactccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     2581 ttttttgctg aaacctcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa
     2641 taaaccggta aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac
     2701 gaccgggtca tcgtggccgg atcttgcggc ccctcggctt gaacgaattg ttagacatta
     2761 tttgccgact accttggtga tctcgccttt cacgtagtgg acaaattctt ccaactgatc
     2821 tgcgcgcgag gccaagcgat cttcttcttg tccaagataa gcctgtctag cttcaagtat
     2881 gacgggctga tactgggccg gcaggcgctc cattgcccag tcggcagcga catccttcgg
     2941 cgcgattttg ccggttactg cgctgtacca aatgcgggac aacgtaagca ctacatttcg
     3001 ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat ttagcgcctc
     3061 aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc gccgctggac ctaccaaggc
     3121 aacgctatgt tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga tcgtggctgg
     3181 ctcgaagata cctgcaagaa tgtcattgcg ctgccattct ccaaattgca gttcgcgctt
     3241 agctggataa cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt ctacagcgcg
     3301 gagaatctcg ctctctccag gggaagccga agtttccaaa aggtcgttga tcaaagctcg
     3361 ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat cactgtgtgg
     3421 cttcaggccg ccatccactg cggagccgta caaatgtacg gccagcaacg tcggttcgag
     3481 atggcgctcg atgacgccaa ctacctctga tagttgagtc gatacttcgg cgatcaccgc
     3541 ttccctcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     3601 gagcggatac atatttgaat gtatttagaa aaataaacaa atagctagct cactcggtcg
     3661 ctacgctccg ggcgtgagac tgcggcgggc gctgcggaca catacaaagt tacccacaga
     3721 ttccgtggat aagcagggga ctaacatgtg aggcaaaaca gcagggccgc gccggtggcg
     3781 tttttccata ggctccgccc tcctgccaga gttcacataa acagacgctt ttccggtgca
     3841 tctgtgggag ccgtgaggct caaccatgaa tctgacagta cgggcgaaac ccgacaggac
     3901 ttaaagatcc ccaccgtttc cggcgggtcg ctccctcttg cgctctcctg ttccgaccct
     3961 gccgtttacc ggatacctgt tccgcctttc tcccttacgg gaagtgtggc gctttctcat
     4021 agctcacaca ctggtatctc ggctcggtgt aggtcgttcg ctccaagctg ggctgtaagc
     4081 aagaactccc cgttcagccc gactgctgcg ccttatccgg taactgttca cttgagtcca
     4141 acccggaaaa gcacggtaaa acgccactgg cagcagccat tggtaactgg gagttcgcag
     4201 aggatttgtt tagctaaaca cgcggttgct cttgaagtgt gcgccaaagt ccggctacac
     4261 tggaaggaca gatttggttg ctgtgctctg cgaaagccag ttaccacggt taagcagttc
     4321 cccaactgac ttaaccttcg atcaaaccac ctccccaggt ggttttttcg tttacagggc
     4381 aaaagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctactgaac
     4441 cgctctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc agccccatac
     4501 gatataagtt gtaattctca tgttagtcat gccccgcgcc caccggaagg agctgactgg
     4561 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     4621 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4681 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
     4741 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga
     4801 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg
     4861 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatgt
     4921 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat
     4981 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt
     5041 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc
     5101 gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc
     5161 ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg
     5221 taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa
     5281 ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg
     5341 gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac
     5401 aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg
     5461 cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg
     5521 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
     5581 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
     5641 cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt
     5701 ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg
     5761 ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta
     5821 tgcgactcct gcattaggaa attaatacga ctcactata
//
