LOCUS       sequence-468480-        5077 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Anthoceros agrestis chaperones: Raf1,BSD2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5077)
  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 5077)
  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/BSD2
FEATURES             Location/Qualifiers
     source          1..5077
                     /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      1758..1805
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1976..2767)
                     /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(2768..2859)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2907..3645)
                     /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(3855..4937)
                     /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     4899..4918
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4938..5015)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(5061..5077,1..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 tggcctctgc gcagggggat gagtctaagg cgacctcgca gtcgggagtg
     1501 aagacaaaga gcattctttg catgaactgc acggggaacg gggctgttgt gtgctcccag
     1561 tgtaagggaa aaggcttgaa ttcggtggat cacttcaacg gccggtacaa agcaggccaa
     1621 atttgctggc tgtgcagggg caagaaacag atgctctgcg gagagtgcaa cggtgccgga
     1681 ttcattggtg gctttctgaa tacgcaagac gattagtcga ccaggcatta ctagagtcac
     1741 actccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     1801 ttttgctgaa acctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata
     1861 aaccggtaaa ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga
     1921 ccgggtcatc gtggccggat cttgcggccc ctcggcttga acgaattgtt agacattatt
     1981 tgccgactac cttggtgatc tcgcctttca cgtagtggac aaattcttcc aactgatctg
     2041 cgcgcgaggc caagcgatct tcttcttgtc caagataagc ctgtctagct tcaagtatga
     2101 cgggctgata ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg
     2161 cgattttgcc ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct
     2221 catcgccagc ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa
     2281 atagatcctg ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa
     2341 cgctatgttc tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct
     2401 cgaagatacc tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag
     2461 ctggataacg ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga
     2521 gaatctcgct ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc
     2581 gcgttgtttc atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct
     2641 tcaggccgcc atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat
     2701 ggcgctcgat gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt
     2761 ccctcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     2821 gcggatacat atttgaatgt atttagaaaa ataaacaaat agctagctca ctcggtcgct
     2881 acgctccggg cgtgagactg cggcgggcgc tgcggacaca tacaaagtta cccacagatt
     2941 ccgtggataa gcaggggact aacatgtgag gcaaaacagc agggccgcgc cggtggcgtt
     3001 tttccatagg ctccgccctc ctgccagagt tcacataaac agacgctttt ccggtgcatc
     3061 tgtgggagcc gtgaggctca accatgaatc tgacagtacg ggcgaaaccc gacaggactt
     3121 aaagatcccc accgtttccg gcgggtcgct ccctcttgcg ctctcctgtt ccgaccctgc
     3181 cgtttaccgg atacctgttc cgcctttctc ccttacggga agtgtggcgc tttctcatag
     3241 ctcacacact ggtatctcgg ctcggtgtag gtcgttcgct ccaagctggg ctgtaagcaa
     3301 gaactccccg ttcagcccga ctgctgcgcc ttatccggta actgttcact tgagtccaac
     3361 ccggaaaagc acggtaaaac gccactggca gcagccattg gtaactggga gttcgcagag
     3421 gatttgttta gctaaacacg cggttgctct tgaagtgtgc gccaaagtcc ggctacactg
     3481 gaaggacaga tttggttgct gtgctctgcg aaagccagtt accacggtta agcagttccc
     3541 caactgactt aaccttcgat caaaccacct ccccaggtgg ttttttcgtt tacagggcaa
     3601 aagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tactgaaccg
     3661 ctctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag ccccatacga
     3721 tataagttgt aattctcatg ttagtcatgc cccgcgccca ccggaaggag ctgactgggt
     3781 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
     3841 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     3901 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt
     3961 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
     4021 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
     4081 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatgtcc
     4141 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
     4201 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
     4261 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
     4321 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
     4381 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
     4441 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
     4501 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
     4561 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
     4621 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
     4681 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
     4741 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
     4801 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
     4861 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
     4921 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     4981 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg
     5041 cgactcctgc attaggaaat taatacgact cactata
//
