LOCUS       sequence-318479-        8934 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of N. tabacum Rubisco large subunit (rbcL) and Rubisco 
            chaperone protiens cpn60-alpha, cpn60-beta, and E. coli groES in E. 
            coli. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8934)
  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 8934)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pET-NtC60-alpha-ESNtLC60-beta
FEATURES             Location/Qualifiers
     source          1..8934
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(40..58)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        126..230
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(231..299,300..1091)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(449..468)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1262..1850
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1751..1770
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2004..2021
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2036..2175
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2101..2120)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2261..2283
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2277..2468)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        3147..3165
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3166..3190
                     /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             3205..3227
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4239..4250
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        4885..4903
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4935..4957
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5010..5063
                     /codon_start=1
                     /product="GroES chaperone mobile loop that interacts with 
                     GroEL"
                     /label=S loop
                     /translation="EVETKSAGGIVLTGSAAA"
     promoter        5276..5294
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5295..5319
                     /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             5334..5356
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        6884..6902
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6903..6927
                     /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)."
     primer_bind     complement(8679..8697)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      8693..8740
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 gataagctgt caaacatgag aattcttgaa gacgaaaggg cctcgtgata cgcctatttt
       61 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
      121 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
      181 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
      241 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
      301 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
      361 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
      421 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
      481 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
      541 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
      601 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
      661 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
      721 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
      781 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
      841 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
      901 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
      961 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     1021 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     1081 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     1141 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     1201 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     1261 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     1321 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     1381 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     1441 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     1501 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     1561 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     1621 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     1681 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     1741 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     1801 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     1861 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     1921 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     1981 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     2041 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     2101 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     2161 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     2221 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     2281 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     2341 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     2401 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     2461 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     2521 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     2581 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     2641 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     2701 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     2761 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     2821 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     2881 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag
     2941 gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg agatggcgga cgcgatggat
     3001 atgttctgcc aagggttggt ttgcgcattc acagttctcc gcaagaattg attggctcca
     3061 attcttggag tggtgaatcc gttagcgagg tgccgccggc cacgatgcgt ccggcgtaga
     3121 ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga
     3181 taacaattcc cctctagcaa taattttgtt taactttaag aaggagatat acatatggct
     3241 aaagaaattg cctttgacca gaaatctagg agtgcccttc aggctggtat cgataagctc
     3301 gctgatgctg tcggtctcac tcttggtccc agggggagga atgtagtgtt ggatgaatat
     3361 ggcaccccaa aggtggttaa tgacggagtt acaattgctc gtgctataga gctacctgat
     3421 gctatggaaa atgctggcgc tgcccttatc agggaggttg caagcaaaac taacgattca
     3481 gccggtgatg gaaccacgac tgcatctgtt cttgctcgcg aaatcattaa acttggtcta
     3541 ttgagtgtta catctggtgc aaatccagtg tctttgaaga agggcattga caagactgta
     3601 ctgggtttga ttgaagagct agaaaagaag gctagacctg ttaaaggtcg tgatgacatc
     3661 aaagctattg cttcgatctc tgctggaaat gatgaaagca ttggcaccat gattgcggat
     3721 gcaattgaca aagtcggtcc agatggtgtg ttgtccatcg agtcatcctc ctcctttgag
     3781 acaactgttc atgttgaaga aggaatggag attgatagag gatatatttc cccacaattt
     3841 gtcaccaacc ctgagaaatt aattgttgag tttgagaatg ctagagtctt agttacggat
     3901 cagaagatct cggctatcaa ggatattatc cccctattgg aaaagacaac tcagttacgc
     3961 tctcctctgc tcattattgc tgaggatgtc accggggaag ctctggctac tcttgtcgtg
     4021 aacaagctgc ggggtatact gaatgttgcc gccatcaaag ctcctggatt tggtgaaagg
     4081 aggaaggctc ttctgcaaga tattgccatt gtgacagggg ctgagtacca agcaactgac
     4141 ctgggcctgc tcattgaggg taccccagtt gaagcacttg gaattgccag aaaggtgacc
     4201 attaccaagg attcaacaac catcattgct gatgcagcat cgaaggacga gatacaggct
     4261 aggattgctc agctcaaaaa ggagttgtcc gagacagact cggtttatga ctccgagaaa
     4321 cttgctgaga gaatcgccaa gttgtctggg ggtgttgccg tcatcaaggt cggagctgct
     4381 acagaggctg agcttgagga ccgcaagctt cgcatcgagg atgcaaagaa tgcaactttt
     4441 gctgcaatcg aagagggaat agtacctggt ggtggtgctg cctttgttca tctatcaact
     4501 tatgtccctg ccattaagga caagctcgaa gatgcagatg aaaagttggg agctgacatc
     4561 attcaaaggg cattagtagc accagcatct ttgatagccc aaaatgctgg aattgaaggg
     4621 gaagtagtag tggagaaggt aagggaagct gaatgggaga tgggttataa cgcgatgaca
     4681 gacaaatacg agaatcttgt tgatgctgga gtgatcgatc cagccaaggt gacaagatgt
     4741 gcattgcaga attcagcatc agttgcagga atggtactta caacacaagc cattgtggtc
     4801 gagaaggcaa agcctaaggc agctgcacct gctgctccac aagggctctc agtgtgatac
     4861 ttggcgcgcg gataacaatt gccataatac gactcactat agggagacca caacggtttc
     4921 cctctagaaa taattttgtt taactttaag aaggagatat acatatgaat attcgtccat
     4981 tgcatgatcg cgtgatcgtc aagcgtaaag aagttgaaac taaatctgct ggcggcatcg
     5041 ttctgaccgg ctctgcagcg gctaaatcca cccgcggcga agtgctggct gtcggcaatg
     5101 gccgtatcct tgaaaatggc gaagtgaagc cgctggatgt gaaagttggc gacatcgtta
     5161 ttttcaacga tggctacggt gtgaaatctg agaagatcga caatgaagaa gtgttgatca
     5221 tgtccgaaag cgacattctg gcaattgttg aagcgtaata cttggcgcgc gaagttaata
     5281 cgactcacta taggggaatt gtgagcggat aacaattccc ctctagaaat aattttgttt
     5341 aactttaaga aggagatata catacccatg tcaccacaaa cagagactaa agcaagtgtt
     5401 ggattcaaag ctggtgttaa agagtacaaa ttgacttatt atactcctga gtaccaaacc
     5461 aaggatactg atatattggc agcattccga gtaactcctc aacctggagt tccacctgaa
     5521 gaagcagggg ccgcggtagc tgccgaatct tctactggta catggacaac tgtatggacc
     5581 gatggactta ccagccttga tcgttacaaa gggcgatgct accgcatcga gcgtgttgtt
     5641 ggagaaaaag atcaatatat tgcttatgta gcttaccctt tagacctttt tgaagaaggt
     5701 tctgttacca acatgtttac ttccattgta ggtaacgtat ttgggttcaa agccctgcgc
     5761 gctctacgtc tggaagatct gcgaatccct cctgcttatg ttaaaacttt ccaaggtccg
     5821 cctcatggga tccaagttga aagagataaa ttgaacaagt atggtcgtcc cctgttggga
     5881 tgtactatta aacctaaatt ggggttatct gctaaaaact acggtagagc tgtttatgaa
     5941 tgtcttcgcg gtggacttga ttttaccaaa gatgatgaga acgtgaactc acaaccattt
     6001 atgcgttgga gagatcgttt cttattttgt gccgaagcac tttataaagc acaggctgaa
     6061 acaggtgaaa tcaaagggca ttacttgaat gctactgcag gtacatgcga agaaatgatc
     6121 aaaagagctg tatttgctag agaattgggc gttccgatcg taatgcatga ctacttaacg
     6181 gggggattca ccgcaaatac tagcttggct cattattgcc gagataatgg tctacttctt
     6241 cacatccacc gtgcaatgca tgcggttatt gatagacaga agaatcatgg tatccacttc
     6301 cgggtattag caaaagcgtt acgtatgtct ggtggagatc atattcactc tggtaccgta
     6361 gtaggtaaac ttgaaggtga aagagacata actttgggct ttgttgattt actgcgtgat
     6421 gattttgttg aacaagatcg aagtcgcggt atttatttca ctcaagattg ggtctcttta
     6481 ccaggtgttc tacccgtggc ttcaggaggt attcacgttt ggcatatgcc tgctctgacc
     6541 gagatctttg gggatgattc cgtactacag ttcggtggag gaactttagg acatccttgg
     6601 ggtaatgcgc caggtgccgt agctaatcga gtagctctag aagcatgtgt aaaagctcgt
     6661 aatgaaggac gtgatcttgc tcaggaaggt aatgaaatta ttcgcgaggc ttgcaaatgg
     6721 agcccggaac tagctgctgc ttgtgaagta tggaaagaga tcgtatttaa ttttgcagca
     6781 gtggacgttt tggataagta atacttggcg cgcccagtgc ggccgcataa tgcttaagtc
     6841 gaacagaaag taatcgtatt gtacacggcc gcataatcga aattaatacg actcactata
     6901 ggggaattgt gagcggataa caattcccca tcttagtata ttagttaagt ataagaagga
     6961 gatatacata tacatatggc agcaaaggag ttgtatttca acaaggatgg atcagctatc
     7021 aagaggctgc aggttggtgt gaacaaactc gctgatctag ttggagttac tcttggccca
     7081 aaaggcagga atgttgttct tgagagcaag tatggtgctc caaagattgt caatgatgga
     7141 gttactgttg ccagagaggt tgagttagag gatccagtag aaaacattgg tgctaaacta
     7201 gtgagacaag ctgctgccaa aaccaatgat ttggctggtg atgggactac aacatctgtc
     7261 gtactagccc agggcctggt tgctgaaggt gttaaggtgg ttgcagctgg tgcaaaccct
     7321 gtcctgatca ccagaggaat tgagaagacc acaaaagcct tagtagctga attgaagaat
     7381 atgtcaaaag aggtagaaga cagtgaatta gcagatgttg ctgcagtaag tgctgggaac
     7441 aactctgaag tagggagtat gattgctgaa gccatgagca aggttggaag gaaaggtgtt
     7501 gtgactcttg aagagggaaa aagtgctgaa aacagtctcc gtgtggttga aggaatgcaa
     7561 tttgaccgtg gttacatctc accatacttc gttactgata gtgagaaaat gtccgttgag
     7621 tatgagaact gtaagttatt actggttgac aagaagataa caaatgcaag agatcttgtt
     7681 ggtgtcctgg aagatgctat cagaaatggt tacccaattt taattattgc cgaagatatt
     7741 gagcaagaag cactagccac ccttgttgtc aataagctta gaggtgcttt aaaggttgct
     7801 gcccttaaag ctcctggttt tggtgaacga aaaagccagt atcttgatga catagcaacc
     7861 cttactggag gcactgtgat tagggaggaa cttggcctta ccttagacaa ggctgacaag
     7921 gaagttctag gccatgcttc taaggtagtg ctgactaagg atgccaccac aattgttggt
     7981 gatggtagca ctcaggaagc agtcaataag cgtgttgtac agattaaaaa cctcatagag
     8041 gcggcagatc aagattatga aaaagaaaag ctaaatgaaa gaattgctaa attatcagga
     8101 ggtgtggctg ttatacaggt tggagcacaa actgaaactg aattgaagga aaagaagctg
     8161 agggtggaag atgctctcaa tgcaacaaag gcagctgttg aggaaggtat tgttgttggt
     8221 ggagggtgca cactcctgcg acttgcttct aaagttgatg ccatcaagga gactcttgag
     8281 aatgatgagg agaaggttgg agctgatatt gttaaaagag cgttgagcta cccattgaag
     8341 ttgattgcta aaaatgctgg tgttaatgga agtgttgtca gtgagaaggt gctctctagt
     8401 gacaatccaa aatttggata caatgctgca actggcaatt atgaagactt gatggctgct
     8461 ggtattattg acccaacaaa ggtggtgaga tgttgtctag agcatgcgtc atcagtggcg
     8521 aagacattct tgatgtcaga ttgtgtggtg gtagagatca aggagcctga accagcagtt
     8581 gctggcaacc caatggacaa ttcaggttat ggatactaat tatgttatat ctttgtaatc
     8641 tgcaacgtac cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     8701 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
     8761 cggatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc tacagcatcc
     8821 agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca cggtgcctga
     8881 ctgcgttagc aatttaactg tgataaacta ccgcattaaa gctagcttat cgat
//
