LOCUS       sequence-457193-        5520 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Notothylas orbicularis Rubisco large and small subunit (C 
            terminus hexahistidine).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5520)
  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 5520)
  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_NoRbcLS
FEATURES             Location/Qualifiers
     source          1..5520
                     /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)."
     CDS             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        169..187
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    188..212
                     /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             246..268
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             1716..1738
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2103..2120
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2139..2157)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2153..2200
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2430..3245)
                     /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(2430..3245)
                     /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     3153..3172
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3246..3337)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3353..4102)
                     /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(3363..3380)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3603..3622)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4298..5380)
                     /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     5342..5361
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5381..5458)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(5504..5520,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcgag
      121 ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa tgcttaagta atacgactca
      181 ctatagggga attgtgagcg gataacaatt cccctgtagg atcccaattc ccctctagaa
      241 ataattttgt ttaactttaa gaaggagata tacatatgtc accacaaacg gagactaaag
      301 caggtgttgg atttaaagct ggtgttaagg attatagatt aacctactat actcctgatt
      361 atgaaaccaa ggatactgat attttggcag cgtttcgaat gactcctcaa ccaggggtgc
      421 cacctgaaga agcaggagcc gcagtagcgg ctgaatcttc aacaggtaca tggaccactg
      481 tttggactga tggacttacc agtcttgatc gttataaagg tcgatgctat gacattgaac
      541 ctgttgctgg agaagagaat caatacattg cttatgttgc ttatccttta gatctctttg
      601 aggaaggttc tgttaccaac atgttcacat ccatcgtagg taatgttttt ggatttaaag
      661 ctttacgagc cttgcgtctg gaagatttaa gaatcccacc tgcttattcc aaaactttcc
      721 aaggtccacc tcatggtatt caggttgaaa gagataaatt gaataaatat ggccgtcctt
      781 tattaggatg taccattaaa ccaaaattag gtttatctgc taaaaactat ggtagagctg
      841 tatatgaatg tcttcgtggc ggacttgatt tcactaagga tgatgagaat gttaattctc
      901 aacctttcat gcgttggaga gatcgtttct tattcgtagc agaagctctt tttaaatctc
      961 aagctgaaac aggcgagatt aaggggcatt acttgaatgc aactgcaggt acgtgtgaag
     1021 aaatgctgaa aagggcaata tttgctagag aattaggggt gcctattata atgcacgact
     1081 atttgacagg aggttttact gcaaatacga gtttggctca ctattgtaga gataatggtt
     1141 tacttctgca cattcaccgt gctatgcatg cagttatcga cagacaaaaa aatcatggta
     1201 ttcactttcg tgtattagct aaagcattac gtatgtctgg tggagatcat attcattcag
     1261 gtactgtcgt aggtaaacta gagggagaac gcgaagtaac tttaggcttt gttgatttac
     1321 ttcgtgacga ttacatcgaa aaagatcgaa gtcgtggtat ctatttcact caagactggg
     1381 tatctatgcc tggtgttctg cctgtcgctt caggaggtat tcatgtctgg catatgcctg
     1441 ctttaactga gattttcgga gatgattctg tattacaatt tggtggggga accctagggc
     1501 acccttgggg aaatgcacct ggtgcagtag ctaatagagt cgccttagaa gcgtgtgtgc
     1561 aagctcgtaa tgagggacgt gatcttgctc gccaaggtaa tgaaattatt cgcgaagctt
     1621 gtaagtggag tccggaatta gcagctgctt gcgaagtatg gaaagaaatt aaattcgaat
     1681 ttgaaacaat tgatactcta taagctagaa ataattttgt ttaactttaa gaaggagata
     1741 tacatatgca ggtgtggaac cccattaacg cacccaagtt cgagaccctg tcctacctgc
     1801 ccaacctgag gcccgatgcc gtcgccaagc agattgacta catgatcagg aatggctgga
     1861 ttccctgcct cgagttcgac gagatcggca ctatccgacg ggatttccac agcagccccg
     1921 ggtactatga tggccgatac tggacaatgt ggaagctccc catgttcggg tgcaccgact
     1981 cagctgcagt tttgagagaa atcgagaact gcaagaacga gtaccccggg tgctacatcc
     2041 gtgtgctagg atttgacagg ctgaggcagg tgcagtgcgt tagcttcatc gtacacaagc
     2101 ctcatcacca tcatcaccac taacctaggc tgctgccacc gctgagcaat aactagcata
     2161 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaacctc aggcatttga
     2221 gaagcacacg gtcacactgc ttccggtagt caataaaccg gtaaaccagc aatagacata
     2281 agcggctatt taacgaccct gccctgaacc gacgacaagc tgacgaccgg gtctccgcaa
     2341 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2401 caaatatgta tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac
     2461 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     2521 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     2581 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     2641 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc
     2701 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     2761 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcggtcgctg
     2821 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     2881 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg
     2941 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     3001 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     3061 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     3121 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     3181 tcagcatcca tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg
     3241 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     3301 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gcatgcagcg ctcttccgct
     3361 tcctcgctca ctgactcgct acgctcggtc gttcgactgc ggcgagcggt gtcagctcac
     3421 tcaaaagcgg taatacggtt atccacagaa tcaggggata aagccggaaa gaacatgtga
     3481 gcaaaaagca aagcaccgga agaagccaac gccgcaggcg tttttccata ggctccgccc
     3541 ccctgacgag catcacaaaa atcgacgctc aagccagagg tggcgaaacc cgacaggact
     3601 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3661 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3721 ctcacgctgt tggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3781 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3841 cccggtaaga cacgacttat cgccactggc agcagccatt ggtaactgat ttagaggact
     3901 ttgtcttgaa gttatgcacc tgttaaggct aaactgaaag aacagatttt ggtgagtgcg
     3961 gtcctccaac ccacttacct tggttcaaag agttggtagc tcagcgaacc ttgagaaaac
     4021 caccgttggt agcggtggtt tttctttatt tatgagatga tgaatcaatc ggtctatcaa
     4081 gtcaacgaac agctattccg ttactctaga tttcagtgca atttatctct tcaaatgtag
     4141 cacctgaagt cagccccata cgatataagt tgtaattctc atgttagtca tgccccgcgc
     4201 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc
     4261 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4321 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4381 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct
     4441 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc
     4501 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt
     4561 cgtatcccac taccgagatg tccgcaccaa cgcgcagccc ggactcggta atggcgcgca
     4621 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat
     4681 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg
     4741 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg
     4801 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca
     4861 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg
     4921 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa
     4981 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa
     5041 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca
     5101 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt
     5161 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt
     5221 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg
     5281 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac
     5341 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac
     5401 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg
     5461 ggatctcgac gctctccctt atgcgactcc tgcattagga aattaatacg actcactata
//
