LOCUS       sequence-457204-        5523 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Phaeoceros carolinianus Rubisco large and small subunit (C
            terminus hexahistidine).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5523)
  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 5523)
  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_PcRbcLS
FEATURES             Location/Qualifiers
     source          1..5523
                     /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             2106..2123
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2142..2160)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2156..2203
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2433..3248)
                     /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(2433..3248)
                     /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     3156..3175
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3249..3340)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3356..4105)
                     /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(3366..3383)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3606..3625)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4301..5383)
                     /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     5345..5364
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5384..5461)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(5507..5523,1..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 cagctgaaga agcaggagcc gcagtagctg ctgaatcttc aacaggtaca tggactactg
      481 tttggactga tggacttact agtcttgatc gttataaagg tcgatgctat gacatcgaac
      541 ctgttgctgg agaagagaat caatacattg cttatgttgc ttatccttta gatctttttg
      601 aggaaggttc tgttaccaac atgttcacat ccatcgtagg taatgttttt ggatttaaag
      661 ctttacgagc cttgcgtctg gaagatttac gaatcccacc tgcttattca aaaactttcc
      721 aaggtccacc tcatggtatt caggttgaaa gggataaatt gaataaatat ggtcgtcctt
      781 tattaggatg taccattaaa ccaaaattag gtttatctgc taaaaactat ggtagagctg
      841 tatatgaatg tcttcgtggc ggacttgatt tcactaagga tgatgagaat gttaattctc
      901 aacctttcat gcgttggaga gatcgtttct tgttcgtagc agaagctatt tttaaatctc
      961 aagctgaaac aggcgagatt aaggggcatt acttgaatgc gactgcaggt acgtgtgagg
     1021 aaatgatgaa aagggcacat tttgctagag aattaggggt gcctattgta atgcacgact
     1081 atttgacagg aggttttact gcaaatacga gtttggctca ctactgtcga gataatggtt
     1141 tacttctcca cattcatcgt gctatgcatg cagttatcga cagacaaaga aatcatggta
     1201 ttcactttcg tgtattagct aaagcattac gtatgtctgg tggagatcat attcattcag
     1261 gtactgtcgt aggtaaatta gagggagaac gtgaagtaac tttaggcttt gttgatttac
     1321 ttcgcgacga ttacatcgaa aaagatcgaa gtcgtggtat ctattttact caagactggg
     1381 tatctatgcc tggtgttttg cctgtcgctt caggaggtat tcatgtctgg catatgcctg
     1441 ctttaactga gattttcgga gatgattctg tattacaatt tggtggggga actctagggc
     1501 acccttgggg aaatgcacct ggtgcagtag ctaatcgagt tgccttagaa gcctgtgtgc
     1561 aagctcgtaa tgagggacgt gatcttgctc gccaaggtaa tgaaattatt cgcgaagctt
     1621 gtaagtggag tccagaatta gcagctgctt gcgaagtatg gaaagaaatt aaattcgaat
     1681 ttgatacaat tgatactcta taagctagaa ataattttgt ttaactttaa gaaggagata
     1741 tacatatgca ggtttggaga cccattgaca accccaggtt cgagactctg tcgtacttgc
     1801 cccctctcac ttccgaccag atcgccaagg agatcgacta catgttgctc aacaagtgga
     1861 ttccttgcct cgagttcgac ccagttggag tcatctcccg ccagtaccac accagccccg
     1921 gatactatga tggccgatac tggacaatgt ggaagttgcc catgttcgga tgcaccgatt
     1981 ctagttctgt gttgagggaa attaataact gcaagaggga gtacccccaa tgcttcatcc
     2041 gtgtgctggg attcgacaac agcaggcagg tgcagtgcgt ctccttcatc gtccacaagc
     2101 cttaccatca ccatcatcac cactaaccta ggctgctgcc accgctgagc aataactagc
     2161 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaac ctcaggcatt
     2221 tgagaagcac acggtcacac tgcttccggt agtcaataaa ccggtaaacc agcaatagac
     2281 ataagcggct atttaacgac cctgccctga accgacgaca agctgacgac cgggtctccg
     2341 caagtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     2401 attcaaatat gtatccgctc atgaattaat tcttagaaaa actcatcgag catcaaatga
     2461 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt
     2521 aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct
     2581 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
     2641 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta
     2701 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
     2761 gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcggtcg
     2821 ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
     2881 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc
     2941 ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg
     3001 gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca
     3061 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
     3121 aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
     3181 aaatcagcat ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata
     3241 tggctcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3301 agcggataca tatttgaatg tatttagaaa aataaacaaa taggcatgca gcgctcttcc
     3361 gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc ggtgtcagct
     3421 cactcaaaag cggtaatacg gttatccaca gaatcagggg ataaagccgg aaagaacatg
     3481 tgagcaaaaa gcaaagcacc ggaagaagcc aacgccgcag gcgtttttcc ataggctccg
     3541 cccccctgac gagcatcaca aaaatcgacg ctcaagccag aggtggcgaa acccgacagg
     3601 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3661 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3721 tagctcacgc tgttggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3781 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3841 caacccggta agacacgact tatcgccact ggcagcagcc attggtaact gatttagagg
     3901 actttgtctt gaagttatgc acctgttaag gctaaactga aagaacagat tttggtgagt
     3961 gcggtcctcc aacccactta ccttggttca aagagttggt agctcagcga accttgagaa
     4021 aaccaccgtt ggtagcggtg gtttttcttt atttatgaga tgatgaatca atcggtctat
     4081 caagtcaacg aacagctatt ccgttactct agatttcagt gcaatttatc tcttcaaatg
     4141 tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgttag tcatgccccg
     4201 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg
     4261 tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4321 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4381 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc
     4441 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
     4501 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
     4561 cgtcgtatcc cactaccgag atgtccgcac caacgcgcag cccggactcg gtaatggcgc
     4621 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
     4681 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
     4741 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
     4801 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
     4861 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
     4921 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
     4981 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
     5041 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
     5101 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
     5161 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
     5221 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
     5281 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
     5341 caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt
     5401 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt
     5461 ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaaattaat acgactcact
     5521 ata
//
