LOCUS       sequence-318839-        7445 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of N. tabacum Rubisco small subunit (rbcS-S1), Rubisco 
            chaperone protiens rbcX, raf1 and A. thaliana rubisco chaperone 
            proteins raf2, and bsd2 in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7445)
  AUTHORS   Lin MT, Stone WD, Chaudhari V, Hanson MR
  TITLE     Small subunits can determine enzyme kinetics of tobacco Rubisco 
            expressed in Escherichia coli.
  JOURNAL   Nat Plants. 2020 Oct;6(10):1289-1299. doi: 
            10.1038/s41477-020-00761-5. Epub 2020 Sep 14.
  PUBMED    32929197
REFERENCE   2  (bases 1 to 7445)
  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: 
            "10.1038/s41477-020-00761-5"; journalName: "Nat Plants"; date: 
            "2020-10"; volume: "6"; issue: "10"; pages: "1289-1299"
COMMENT     Sequence Label: pCDF-NtXNtS1R1AtR2B2
FEATURES             Location/Qualifiers
     source          1..7445
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        455..473
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    474..498
                     /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)."
     promoter        1851..1869
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1870..1894
                     /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             1909..1931
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2458..2487
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="ASMTGGQQMG"
     terminator      2555..2602
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2769..2787
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2788..2812
                     /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             2827..2849
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3106..3135
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="ASMTGGQQMG"
     terminator      3203..3250
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             3410..3454
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     terminator      3506..3553
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3724..4515)
                     /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(4516..4607)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4655..5393)
                     /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(5603..6685)
                     /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     6647..6666
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6686..6763)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        6809..6827
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6828..6852
                     /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)."
     promoter        7366..7384
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             7416..7438
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 atgcaggtgt ggccaccaat taacaagaag aagtacgaga ctctctcata ccttcctgat
       61 ttgagccagg agcaattgct tagtgaagtt gagtaccttt tgaaaaatgg atgggttcct
      121 tgcttggaat tcgagactga gcacggattt gtctatcgtg aaaacaacaa gtcaccagga
      181 tactatgatg gcagatactg gaccatgtgg aagctaccta tgttcggatg cactgatgcc
      241 acccaagtgt tggctgaggt agaagaggcg aagaaggcat acccacaggc ctggatccgt
      301 atcattggat tcgacaacgt gcgtcaagtg cagtgcatca gtttcattgc ctacaagcct
      361 gaaggatatt aatacttggc gcgcccagtg cggccgcata atgcttaagt cgaacagaaa
      421 gtaatcgtat tgtacacggc cgcataatcg aaattaatac gactcactat aggggaattg
      481 tgagcggata acaattcccc atcttagtat attagttaag tataagaagg agatatacat
      541 atgcaacaac tttatcagcc ttttagacct cctcctccac ctctcccacc caaattccgc
      601 aatcttgaca caaatgcaaa gctcgaagtt ctatctaatc gccttggtct ttggtatgaa
      661 tatgctcctt tgatacctta tttaacaagt gaagggttta ctccttcaac tcttgaggaa
      721 atcactggcc ttactggagt tgaacagaat cggttggttg ttgctgccca agttcgtgat
      781 actcttgttg agtctgctgc tttagatgaa gaaacgttgt cgtattttga atctggaggt
      841 gctgaattat tatatgaaat tcgtttgctt agtgctaggc aacgaactga tgcagcaaca
      901 tttttagtga agaatgggtt tgatgcaaaa caagctcaag atttagctag agcaattaaa
      961 gattatccgc gtagacgtgt ggattatggt tgggacaagt ttaatggtga ttctcctggt
     1021 gattgtttag cttttatgta ctttagatta gctcaagaat atgctgcggc agcttctgag
     1081 gatttgagga ggtcgtcaat ggagaaggca ttagaagttg ttgagtctga aagtgcgagg
     1141 aatttgcttg tgatggagct agaaggacgt gaggttgcta aggagagtgt acttgatgat
     1201 ggggtgacag tgcctttggt gagaatgaag ttaggggagg tagctgagtc tacgattgta
     1261 gtcgttttgc ctgtttgtaa ggcggaggga agggacgtgg aggtggaggc ggcgccgtgg
     1321 gaatgtggtg gcgtagggga ttttggtata gtggaggcgg agaaggattg gaggagatgg
     1381 gttgttttgc ccggatggca acctatcgcg gggttagaga gaggtggagt tgcagtgtcg
     1441 ttcaagagcg ggaacttttt gccgtggagg gagaagagta agtacaagca agagccggtt
     1501 ttggtagtgg cggatagggg aaggacagaa gtggcgtcgg aggacgggtt ttacttggtg
     1561 gttgatggtg gagacgggag caatgaggag ggattgaaag tggaaagagg gtcaacattg
     1621 aagaaaagag gggttgaaca gagcttagga attgtgctta tagtagttag gccaccaagg
     1681 tgggaagatg aagaacaatt aggtgaagaa gattgggatt aaagaccaag aagcttagct
     1741 ttctagactg caggtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
     1801 cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat taatacgact
     1861 cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt
     1921 taagaaggag atatacatat gatgtctaat ctggcgcagg attttcttgg tgacttcgga
     1981 gctcgtgacc cttacccgga ggagatagcg agtcaattcg gagataaagt gttgggatgc
     2041 caaagcactg agcacaagat tttgatacca aacgcatctg ttttgtctct ctcccagctt
     2101 cagtgttccc ctgtttcgtc ttcacagcct cctttgtccg gcgatgatgc cagaactctc
     2161 ctccacaagg ttttgggatg gagtatagtg gataatgaag cgggtggtct gaaaataagg
     2221 tgtatgtgga aagtgaggga ttttgggtgc ggtgttgaac tcataaacag gatccataag
     2281 gttgctgaag cttctggtca ttacccttct cttcacttgg aaagtcctac ccaagttcga
     2341 gctgaactat ttacctcttc tatcggaggg ttgagcatga atgatttcat aatggcggct
     2401 aaaatagatg atatcaagac ttctgatctt tcccctagga aaagagcttg ggcgtgagct
     2461 agcatgactg gtggacagca aatgggtcgc ggatccggct gctaacaaag cccgaaagga
     2521 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     2581 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatatc ccgcaagagg
     2641 cccggcagta ccggcataac caagcactag tgtcggcgat ataggcgcca gcaaccgcac
     2701 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc
     2761 cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga
     2821 aataattttg tttaacttta agaaggagat atacatatga tggccgcaaa caataatcct
     2881 cagggcacta aaccaaacag tttagtttgt gcaaattgtg aaggggaagg ttgtgtggca
     2941 tgttctcaat gcaaaggagg tggtgtgaat ttgattgatc atttcaatgg tcagtttaaa
     3001 gctggtgcac tctgttggct ttgcagaggt aaaaaggagg tgttgtgtgg ggactgcaat
     3061 ggagctggtt tcattggtgg gttcttgagc accttcgatg agtaggctag catgactggt
     3121 ggacagcaaa tgggtcgcag atccggctgc taacaaagcc cgaaaggaag ctgagttggc
     3181 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     3241 gggttttttg ctgaaaggag gaactatatc cggatatccc gcaagaggcc cggcagtacc
     3301 ggcataacca agcggtacca gcggtcaatt gtagtggagg tgctagcgca tgcggaattg
     3361 gatatcggcc ggccacgcga tcgctgacgt cggtaccctc gagtctggta aagaaaccgc
     3421 tgctgcgaaa tttgaacgcc agcacatgga ctcgtctact agcgcagctt aattaaccta
     3481 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     3541 gaggggtttt ttgctgaaac ctcaggcatt tgagaagcac acggtcacac tgcttccggt
     3601 agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac cctgccctga
     3661 accgacgacc gggtcatcgt ggccggatct tgcggcccct cggcttgaac gaattgttag
     3721 acattatttg ccgactacct tggtgatctc gcctttcacg tagtggacaa attcttccaa
     3781 ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca agataagcct gtctagcttc
     3841 aagtatgacg ggctgatact gggccggcag gcgctccatt gcccagtcgg cagcgacatc
     3901 cttcggcgcg attttgccgg ttactgcgct gtaccaaatg cgggacaacg taagcactac
     3961 atttcgctca tcgccagccc agtcgggcgg cgagttccat agcgttaagg tttcatttag
     4021 cgcctcaaat agatcctgtt caggaaccgg atcaaagagt tcctccgccg ctggacctac
     4081 caaggcaacg ctatgttctc ttgcttttgt cagcaagata gccagatcaa tgtcgatcgt
     4141 ggctggctcg aagatacctg caagaatgtc attgcgctgc cattctccaa attgcagttc
     4201 gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc acaacaatgg tgacttctac
     4261 agcgcggaga atctcgctct ctccagggga agccgaagtt tccaaaaggt cgttgatcaa
     4321 agctcgccgc gttgtttcat caagccttac ggtcaccgta accagcaaat caatatcact
     4381 gtgtggcttc aggccgccat ccactgcgga gccgtacaaa tgtacggcca gcaacgtcgg
     4441 ttcgagatgg cgctcgatga cgccaactac ctctgatagt tgagtcgata cttcggcgat
     4501 caccgcttcc ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4561 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag ctagctcact
     4621 cggtcgctac gctccgggcg tgagactgcg gcgggcgctg cggacacata caaagttacc
     4681 cacagattcc gtggataagc aggggactaa catgtgaggc aaaacagcag ggccgcgccg
     4741 gtggcgtttt tccataggct ccgccctcct gccagagttc acataaacag acgcttttcc
     4801 ggtgcatctg tgggagccgt gaggctcaac catgaatctg acagtacggg cgaaacccga
     4861 caggacttaa agatccccac cgtttccggc gggtcgctcc ctcttgcgct ctcctgttcc
     4921 gaccctgccg tttaccggat acctgttccg cctttctccc ttacgggaag tgtggcgctt
     4981 tctcatagct cacacactgg tatctcggct cggtgtaggt cgttcgctcc aagctgggct
     5041 gtaagcaaga actccccgtt cagcccgact gctgcgcctt atccggtaac tgttcacttg
     5101 agtccaaccc ggaaaagcac ggtaaaacgc cactggcagc agccattggt aactgggagt
     5161 tcgcagagga tttgtttagc taaacacgcg gttgctcttg aagtgtgcgc caaagtccgg
     5221 ctacactgga aggacagatt tggttgctgt gctctgcgaa agccagttac cacggttaag
     5281 cagttcccca actgacttaa ccttcgatca aaccacctcc ccaggtggtt ttttcgttta
     5341 cagggcaaaa gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5401 ctgaaccgct ctagatttca gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc
     5461 ccatacgata taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc ggaaggagct
     5521 gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta
     5581 acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5641 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg
     5701 tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct
     5761 gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga
     5821 tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg
     5881 agatgtccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca
     5941 tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg
     6001 tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt
     6061 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta
     6121 atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca
     6181 gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat
     6241 caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat
     6301 ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg
     6361 ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt
     6421 gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg
     6481 aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg
     6541 gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt
     6601 ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga
     6661 catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct
     6721 atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct
     6781 cccttatgcg actcctgcat taggaaatta atacgactca ctatagggga attgtgagcg
     6841 gataacaatt cccctgtaga aataattttg tttaacttta ataaggagat ataccatggc
     6901 aaaagcattt aagggtaatt acaagaattt aaagaaggag aaaaagagta ggaggaatct
     6961 tgtggttgtc agtgatttag gtggacagta tgaatatggc tttgatgata tcaaaacgca
     7021 aattcttaac tacttcacat ttaaagcagt gaggactatt ttgaaccagc ttaatgaaat
     7081 ggactctcca caatatcaat ggttcaatga ttatgttaca gtaaataagc ttagggatgg
     7141 gaagcgtttt attcgcaacc ttgctaagga gaagcaagaa ctcgctgaaa aagtgatggt
     7201 aacacggctc agtctgtatg cgaaatggat aaaaaaatgt gatcacgagg agatctacaa
     7261 ccgaatatct gatcagaatg tggaagtgat gcgtgagcgg ctcatggaaa ctgtgatttg
     7321 gccatcagat gacacgaact cggagagagt tgggtgaggc gcgcgtaata cgactcacta
     7381 tagggagacc acaacggttt ccctctagaa ataattttgt ttaactttaa gaaggagata
     7441 tacat
//
