LOCUS       sequence-456757-        5667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Anthoceros agrestis Rubisco large and small subunit (C 
            terminus hexahistidine).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5667)
  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 5667)
  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: pRSF_AaRbcLS
FEATURES             Location/Qualifiers
     source          1..5667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(40..1122)
                     /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     1084..1103
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1123..1200)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        1246..1264
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1265..1289
                     /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             1323..1345
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             2793..2815
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3180..3197
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        3314..3332
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3333..3357
                     /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)."
     CDS             3466..3510
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3548..3566)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3562..3609
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3839..4654)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3839..4654)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4562..4581
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4655..4746)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4762..5511)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(4772..4789)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5012..5031)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg
       61 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
      121 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
      181 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
      241 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc
      301 gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg
      361 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
      421 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
      481 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
      541 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
      601 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
      661 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
      721 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
      781 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
      841 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
      901 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
      961 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     1021 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     1081 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
     1141 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc
     1201 cgggatctcg acgctctccc ttatgcgact cctgcattag gaaattaata cgactcacta
     1261 taggggaatt gtgagcggat aacaattccc ctgtaggatc ccaattcccc tctagaaata
     1321 attttgttta actttaagaa ggagatatac atatgtcacc acaaacggag actaaagcag
     1381 gtgttggatt taaagctggt gttaaagatt atagattaac ctattatacc cctgattacg
     1441 agaccaagga tactgatatt ttggcagcgt ttcgaatgac tcctcaacca ggggtgccac
     1501 ctgaagaagc aggagccgca gtagctgctg aatcttcaac tggtacatgg actactgttt
     1561 ggaccgatgg acttaccagc cttgaccgtt acaaaggtcg atgctacgac attgaacctg
     1621 ttgctggaga ggaaaatcaa tatattgctt atgttgctta tccacttgat ttgtttgagg
     1681 agggttctgt taccaacatg ttcacttcca ttgtaggtaa tgtttttgga tttaaagctt
     1741 tacgagcttt acgtctagaa gatttacgaa ttccacccgc ttattccaaa acctttcaag
     1801 gtccacctca tggtattcaa gttgaaaggg ataaattgaa taaatatggt cgtcctttat
     1861 taggatgtac cattaaacca aagttaggtt tatctgccaa aaactatggt agagctgtat
     1921 atgaatgtct tcgtggtgga cttgatttca ccaaagatga cgaaaatgtt aattctcaac
     1981 cttttatgcg ttggagagat cgttttctat tcgtagcaga agctattttt aaatctcaag
     2041 ctgaaacagg agagattaaa gggcattact taaatgccac tgcaggtaca tgtgaggaaa
     2101 tgatgaaaag ggcacaattt gctagagaat taggaatgcc tatcgtaatg catgattatt
     2161 tgacaggagg ttttactgca aatacgaccc tggctcatta ttgtagagac aatggtttac
     2221 ttcttcatat tcatcgtgct atgcatgcag ttattgacag acaaagaaat catggtattc
     2281 actttcgtgt attagctaaa gcattacgta tgtccggggg agatcatatt cattcaggta
     2341 cagtagtagg taaattagag ggagaacgtg aagtaacttt aggttttgtg gatttacttc
     2401 gtgacgatta tattgaaaaa gatagaagtc gtgggatata ttttactcaa gactgggtct
     2461 ctatgcctgg tgttttgcct gtggcttcag gtggtattca tgtttggcat atgccagctt
     2521 taactgagat tttcggagat gattccgtac tacagtttgg cgggggaact ctaggacacc
     2581 cttggggaaa tgcacctggt gcagtagcta atcgagttgc tttagaagct tgtgtgcaag
     2641 ctcgtaatga aggacgtgat cttgctcgcg agggtaatga tattatccgt gaagctagta
     2701 aatggagtcc tgaattagca gctgcttgtg aagtatggaa agaaattaaa ttcgtttttg
     2761 agacgattga tactctataa gctagaaata attttgttta actttaagaa ggagatatac
     2821 atatgcaggt gtggaacccc attgacaacc ccaagttcga gaccctctct tacttgcccc
     2881 ctctgaccga caaccaaata gccagggaga tcgactacat gctccgcaac aagtggatcc
     2941 cctgcctcga gttcgacccg agcggtacca tcaccaccct gcccggccag cccggatact
     3001 atggcggccg atactggaca atgtggaagc tgcccatgtt cggttgcaac aacgctggct
     3061 atgtgctgag ggagattgag cactgcaaaa acgcctaccc cggctgcttc atccgtgtcc
     3121 ttggcttcga caacatcagg caggtgcagt gctgcgcctt catcgtccac aagccccagc
     3181 atcaccatca tcaccactaa agccaggatc cgaattcgag ctcggcgcgc ctgcaggtcg
     3241 acaagcttgc ggccgcataa tgcttaagtc gaacagaaag taatcgtatt gtacacggcc
     3301 gcataatcga aattaatacg actcactata ggggaattgt gagcggataa caattcccca
     3361 tcttagtata ttagttaagt ataagaagga gatatacata tggcagatct caattggata
     3421 tcggccggcc acgcgatcgc tgacgtcggt accctcgagt ctggtaaaga aaccgctgct
     3481 gcgaaatttg aacgccagca catggactcg tctactagcg cagcttaatt aacctaggct
     3541 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     3601 ggttttttgc tgaaacctca ggcatttgag aagcacacgg tcacactgct tccggtagtc
     3661 aataaaccgg taaaccagca atagacataa gcggctattt aacgaccctg ccctgaaccg
     3721 acgacaagct gacgaccgag tctccgcaag tggcactttt cggggaaatg tgcgcggaac
     3781 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga attaattctt
     3841 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
     3901 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
     3961 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
     4021 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
     4081 aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc
     4141 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
     4201 cctgagcgag acgaaatacg cggtcgctgt taaaaggaca attacaaaca ggaatcgaat
     4261 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
     4321 cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat
     4381 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
     4441 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
     4501 actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat
     4561 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
     4621 tagagcaaga cgtttcccgt tgaatatggc tcatactctt cctttttcaa tattattgaa
     4681 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4741 aacaaatagg catgcagcgc tcttccgctt cctcgctcac tgactcgcta cgctcggtcg
     4801 ttcgactgcg gcgagcggtg tcagctcact caaaagcggt aatacggtta tccacagaat
     4861 caggggataa agccggaaag aacatgtgag caaaaagcaa agcaccggaa gaagccaacg
     4921 ccgcaggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4981 agccagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5041 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5101 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgtt ggtatctcag ttcggtgtag
     5161 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5221 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5281 gcagccattg gtaactgatt tagaggactt tgtcttgaag ttatgcacct gttaaggcta
     5341 aactgaaaga acagattttg gtgagtgcgg tcctccaacc cacttacctt ggttcaaaga
     5401 gttggtagct cagcgaacct tgagaaaacc accgttggta gcggtggttt ttctttattt
     5461 atgagatgat gaatcaatcg gtctatcaag tcaacgaaca gctattccgt tactctagat
     5521 ttcagtgcaa tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt
     5581 gtaattctca tgttagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
     5641 ctcaagggca tcggtcgaga tcccggt
//
