LOCUS       sequence-318842-        7421 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of N. tabacum Rubisco small subunit (rbcS-S1), Rubisco 
            chaperone protiens rbcX, raf1, bsd2 and raf2 in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7421)
  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 7421)
  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-NtXNtS1R1R2B2
FEATURES             Location/Qualifiers
     source          1..7421
                     /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             2437..2466
                     /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      2534..2581
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2748..2766
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2767..2791
                     /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             2806..2828
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3082..3111
                     /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      3179..3226
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             3386..3430
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     terminator      3482..3529
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3700..4491)
                     /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(4492..4583)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4631..5369)
                     /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(5579..6661)
                     /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     6623..6642
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6662..6739)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        6785..6803
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6804..6828
                     /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        7342..7360
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             7392..7414
                     /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 gctattgggc gattttgggg cgagagatcc attcccagct
     1981 gaaatagaga gtaaattcgg tgaaaaagtt ttgggtaata ctgacacgga gcacaaaatt
     2041 ctcatcccag ctgcttctgc tctgtccctt gctcagcaag actgcattgt catctcacct
     2101 ggtcaaactc ctctctcgga atatgaagct cgacagctcc tattcaaggt tgttggctgg
     2161 agactggcac atgaagatgg ggtgctcaag ctacaatgta catggaaatt aagagacttc
     2221 gaatgtgggg ttgaacttat caatcgaata ggcaaagtgg tggaaggcac tgggcacgtt
     2281 cccacacttc ttcacttaga gcagtccaat caagttcgtg ctgagctatg gactgcttcc
     2341 atcggaggtt tgagcataaa cgatttcatt gtagctgcta aaatagatca gataaagaca
     2401 tcggaccttg tcccaagaaa aagagtttgg gcataagcta gcatgactgg tggacagcaa
     2461 atgggtcgcg gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     2521 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     2581 gctgaaagga ggaactatat ccggatatcc cgcaagaggc ccggcagtac cggcataacc
     2641 aagcactagt gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg
     2701 ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaattaa tacgactcac
     2761 tataggggaa ttgtgagcgg ataacaattc ccctctagaa ataattttgt ttaactttaa
     2821 gaaggagata tacatatgat ggccactgac agtgataaag acacgaaagt aaggagcata
     2881 gtctgtcaaa attgtgatgg aaatggtgca gtgtcgtgct cacagtgcaa aggctctggg
     2941 gttaactctg tagatcattt caacgggcgg ttcaaagctg gtggactatg ttggttgtgc
     3001 agaggtaaaa aagatgtgtt atgtggtgac tgcaatggtg ctggatttct tggtggattt
     3061 atgagtacgt tcgatgagta ggctagcatg actggtggac agcaaatggg tcgcagatcc
     3121 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     3181 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     3241 tatatccgga tatcccgcaa gaggcccggc agtaccggca taaccaagcg gtaccagcgg
     3301 tcaattgtag tggaggtgct agcgcatgcg gaattggata tcggccggcc acgcgatcgc
     3361 tgacgtcggt accctcgagt ctggtaaaga aaccgctgct gcgaaatttg aacgccagca
     3421 catggactcg tctactagcg cagcttaatt aacctaggct gctgccaccg ctgagcaata
     3481 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaacctca
     3541 ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca
     3601 atagacataa gcggctattt aacgaccctg ccctgaaccg acgaccgggt catcgtggcc
     3661 ggatcttgcg gcccctcggc ttgaacgaat tgttagacat tatttgccga ctaccttggt
     3721 gatctcgcct ttcacgtagt ggacaaattc ttccaactga tctgcgcgcg aggccaagcg
     3781 atcttcttct tgtccaagat aagcctgtct agcttcaagt atgacgggct gatactgggc
     3841 cggcaggcgc tccattgccc agtcggcagc gacatccttc ggcgcgattt tgccggttac
     3901 tgcgctgtac caaatgcggg acaacgtaag cactacattt cgctcatcgc cagcccagtc
     3961 gggcggcgag ttccatagcg ttaaggtttc atttagcgcc tcaaatagat cctgttcagg
     4021 aaccggatca aagagttcct ccgccgctgg acctaccaag gcaacgctat gttctcttgc
     4081 ttttgtcagc aagatagcca gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag
     4141 aatgtcattg cgctgccatt ctccaaattg cagttcgcgc ttagctggat aacgccacgg
     4201 aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg cggagaatct cgctctctcc
     4261 aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag
     4321 ccttacggtc accgtaacca gcaaatcaat atcactgtgt ggcttcaggc cgccatccac
     4381 tgcggagccg tacaaatgta cggccagcaa cgtcggttcg agatggcgct cgatgacgcc
     4441 aactacctct gatagttgag tcgatacttc ggcgatcacc gcttccctca tactcttcct
     4501 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4561 atgtatttag aaaaataaac aaatagctag ctcactcggt cgctacgctc cgggcgtgag
     4621 actgcggcgg gcgctgcgga cacatacaaa gttacccaca gattccgtgg ataagcaggg
     4681 gactaacatg tgaggcaaaa cagcagggcc gcgccggtgg cgtttttcca taggctccgc
     4741 cctcctgcca gagttcacat aaacagacgc ttttccggtg catctgtggg agccgtgagg
     4801 ctcaaccatg aatctgacag tacgggcgaa acccgacagg acttaaagat ccccaccgtt
     4861 tccggcgggt cgctccctct tgcgctctcc tgttccgacc ctgccgttta ccggatacct
     4921 gttccgcctt tctcccttac gggaagtgtg gcgctttctc atagctcaca cactggtatc
     4981 tcggctcggt gtaggtcgtt cgctccaagc tgggctgtaa gcaagaactc cccgttcagc
     5041 ccgactgctg cgccttatcc ggtaactgtt cacttgagtc caacccggaa aagcacggta
     5101 aaacgccact ggcagcagcc attggtaact gggagttcgc agaggatttg tttagctaaa
     5161 cacgcggttg ctcttgaagt gtgcgccaaa gtccggctac actggaagga cagatttggt
     5221 tgctgtgctc tgcgaaagcc agttaccacg gttaagcagt tccccaactg acttaacctt
     5281 cgatcaaacc acctccccag gtggtttttt cgtttacagg gcaaaagatt acgcgcagaa
     5341 aaaaaggatc tcaagaagat cctttgatct tttctactga accgctctag atttcagtgc
     5401 aatttatctc ttcaaatgta gcacctgaag tcagccccat acgatataag ttgtaattct
     5461 catgttagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     5521 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     5581 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5641 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     5701 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     5761 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     5821 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc
     5881 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     5941 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     6001 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     6061 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     6121 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     6181 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     6241 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     6301 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     6361 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     6421 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     6481 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     6541 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     6601 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     6661 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     6721 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
     6781 aaattaatac gactcactat aggggaattg tgagcggata acaattcccc tgtagaaata
     6841 attttgttta actttaataa ggagatatac catggcaaaa gcatttaagg gtaattacaa
     6901 gaatttaaag aaggagaaaa agagtaggag gaatcttgtg gttgtcagtg atttaggtgg
     6961 acagtatgaa tatggctttg atgatatcaa aacgcaaatt cttaactact tcacatttaa
     7021 agcagtgagg actattttga accagcttaa tgaaatggac tctccacaat atcaatggtt
     7081 caatgattat gttacagtaa ataagcttag ggatgggaag cgttttattc gcaaccttgc
     7141 taaggagaag caagaactcg ctgaaaaagt gatggtaaca cggctcagtc tgtatgcgaa
     7201 atggataaaa aaatgtgatc acgaggagat ctacaaccga atatctgatc agaatgtgga
     7261 agtgatgcgt gagcggctca tggaaactgt gatttggcca tcagatgaca cgaactcgga
     7321 gagagttggg tgaggcgcgc gtaatacgac tcactatagg gagaccacaa cggtttccct
     7381 ctagaaataa ttttgtttaa ctttaagaag gagatataca t
//
