LOCUS       sequence-456758-        5286 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Anthoceros agrestis Rubisco large (N terminus 
            hexahistidine).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5286)
  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 5286)
  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_AaNHis_RbcL
FEATURES             Location/Qualifiers
     source          1..5286
                     /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)"
     CDS             1356..1373
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2799..2816
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        2933..2951
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2952..2976
                     /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             3085..3129
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3167..3185)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3181..3228
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3458..4273)
                     /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(3458..4273)
                     /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     4181..4200
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4274..4365)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4381..5130)
                     /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(4391..4408)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4631..4650)
                     /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 atatgcatca ccaccatcac cattcaccac
     1381 aaacggagac taaagcaggt gttggattta aagctggtgt taaagattat agattaacct
     1441 attatacccc tgattacgag accaaggata ctgatatttt ggcagcgttt cgaatgactc
     1501 ctcaaccagg ggtgccacct gaagaagcag gagccgcagt agctgctgaa tcttcaactg
     1561 gtacatggac tactgtttgg accgatggac ttaccagcct tgaccgttac aaaggtcgat
     1621 gctacgacat tgaacctgtt gctggagagg aaaatcaata tattgcttat gttgcttatc
     1681 cacttgattt gtttgaggag ggttctgtta ccaacatgtt cacttccatt gtaggtaatg
     1741 tttttggatt taaagcttta cgagctttac gtctagaaga tttacgaatt ccacccgctt
     1801 attccaaaac ctttcaaggt ccacctcatg gtattcaagt tgaaagggat aaattgaata
     1861 aatatggtcg tcctttatta ggatgtacca ttaaaccaaa gttaggttta tctgccaaaa
     1921 actatggtag agctgtatat gaatgtcttc gtggtggact tgatttcacc aaagatgacg
     1981 aaaatgttaa ttctcaacct tttatgcgtt ggagagatcg ttttctattc gtagcagaag
     2041 ctatttttaa atctcaagct gaaacaggag agattaaagg gcattactta aatgccactg
     2101 caggtacatg tgaggaaatg atgaaaaggg cacaatttgc tagagaatta ggaatgccta
     2161 tcgtaatgca tgattatttg acaggaggtt ttactgcaaa tacgaccctg gctcattatt
     2221 gtagagacaa tggtttactt cttcatattc atcgtgctat gcatgcagtt attgacagac
     2281 aaagaaatca tggtattcac tttcgtgtat tagctaaagc attacgtatg tccgggggag
     2341 atcatattca ttcaggtaca gtagtaggta aattagaggg agaacgtgaa gtaactttag
     2401 gttttgtgga tttacttcgt gacgattata ttgaaaaaga tagaagtcgt gggatatatt
     2461 ttactcaaga ctgggtctct atgcctggtg ttttgcctgt ggcttcaggt ggtattcatg
     2521 tttggcatat gccagcttta actgagattt tcggagatga ttccgtacta cagtttggcg
     2581 ggggaactct aggacaccct tggggaaatg cacctggtgc agtagctaat cgagttgctt
     2641 tagaagcttg tgtgcaagct cgtaatgaag gacgtgatct tgctcgcgag ggtaatgata
     2701 ttatccgtga agctagtaaa tggagtcctg aattagcagc tgcttgtgaa gtatggaaag
     2761 aaattaaatt cgtttttgag acgattgata ctctataaca tcaccatcat caccactaaa
     2821 gccaggatcc gaattcgagc tcggcgcgcc tgcaggtcga caagcttgcg gccgcataat
     2881 gcttaagtcg aacagaaagt aatcgtattg tacacggccg cataatcgaa attaatacga
     2941 ctcactatag gggaattgtg agcggataac aattccccat cttagtatat tagttaagta
     3001 taagaaggag atatacatat ggcagatctc aattggatat cggccggcca cgcgatcgct
     3061 gacgtcggta ccctcgagtc tggtaaagaa accgctgctg cgaaatttga acgccagcac
     3121 atggactcgt ctactagcgc agcttaatta acctaggctg ctgccaccgc tgagcaataa
     3181 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaacctcag
     3241 gcatttgaga agcacacggt cacactgctt ccggtagtca ataaaccggt aaaccagcaa
     3301 tagacataag cggctattta acgaccctgc cctgaaccga cgacaagctg acgaccgagt
     3361 ctccgcaagt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3421 aatacattca aatatgtatc cgctcatgaa ttaattctta gaaaaactca tcgagcatca
     3481 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
     3541 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc
     3601 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
     3661 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
     3721 gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
     3781 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc
     3841 ggtcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg
     3901 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
     3961 ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
     4021 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
     4081 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
     4141 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
     4201 catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt
     4261 gaatatggct catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4321 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggc atgcagcgct
     4381 cttccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg cgagcggtgt
     4441 cagctcactc aaaagcggta atacggttat ccacagaatc aggggataaa gccggaaaga
     4501 acatgtgagc aaaaagcaaa gcaccggaag aagccaacgc cgcaggcgtt tttccatagg
     4561 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gccagaggtg gcgaaacccg
     4621 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4681 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4741 tctcatagct cacgctgttg gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4801 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4861 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccattgg taactgattt
     4921 agaggacttt gtcttgaagt tatgcacctg ttaaggctaa actgaaagaa cagattttgg
     4981 tgagtgcggt cctccaaccc acttaccttg gttcaaagag ttggtagctc agcgaacctt
     5041 gagaaaacca ccgttggtag cggtggtttt tctttattta tgagatgatg aatcaatcgg
     5101 tctatcaagt caacgaacag ctattccgtt actctagatt tcagtgcaat ttatctcttc
     5161 aaatgtagca cctgaagtca gccccatacg atataagttg taattctcat gttagtcatg
     5221 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
     5281 cccggt
//
