LOCUS Exported 7442 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expression of N. tabacum Rubisco small subunit (rbcS-T1), Rubisco chaperone protiens rbcX, raf1, bsd2 and A. thaliana Rubisco chaperone proteins raf2 in E. coli.. ACCESSION . VERSION . KEYWORDS pCDF-NtXNtT1R1AtR2NtB2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7442) AUTHORS Lin MT, Stone WD, Chaudhar Vi, Hanson MR TITLE Small subunits can determine enzyme kinetics of tobacco Rubisco expressed in Escherichia coli JOURNAL Nat Plants REFERENCE 2 (bases 1 to 7442) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7442 /organism="synthetic DNA construct" /mol_type="other DNA" 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 3103..3132 /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 3200..3247 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" CDS 3407..3451 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" terminator 3503..3550 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" CDS complement(3721..4512) /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(4513..4604) /gene="bla" /label=AmpR promoter rep_origin complement(4652..5390) /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(5600..6682) /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 6644..6663 /label=LacI-R /note="LacI, reverse primer" promoter complement(6683..6760) /gene="lacI" /label=lacI promoter promoter 6806..6824 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 6825..6849 /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 7363..7381 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 7413..7435 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" ORIGIN 1 atgcaggtat ggcccccata tggcaagaag aagtacgaaa ctctctcata ccttcccgat 61 ttaagcgagg agcaattgct tagtgaaatt gagtaccttt tgaaaaatgg atgggttcct 121 tgtttggaat tcgagactga gcgcggattt gtctaccgtg aaaacaacaa gtcaccagga 181 tactatgacg gcagatactg gaccatgtgg aagctgccca tgtttgggtg cactgatgcc 241 acccaagtgt tagctgaggt gggagaggcg aagaaggcat acccagaggc ctggatccgt 301 atcattggat tcgacaacgt gcgtcaagtg cagtgcatca gtttcattgc ctacaagcct 361 gaaggctact 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 tggccactga cagtgataaa 2881 gacacgaaag taaggagcat agtctgtcaa aattgtgatg gaaatggtgc agtgtcgtgc 2941 tcacagtgca aaggctctgg ggttaactct gtagatcatt tcaacgggcg gttcaaagct 3001 ggtggactat gttggttgtg cagaggtaaa aaagatgtgt tatgtggtga ctgcaatggt 3061 gctggatttc ttggtggatt tatgagtacg ttcgatgagt aggctagcat gactggtgga 3121 cagcaaatgg gtcgcagatc cggctgctaa caaagcccga aaggaagctg agttggctgc 3181 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg 3241 ttttttgctg aaaggaggaa ctatatccgg atatcccgca agaggcccgg cagtaccggc 3301 ataaccaagc ggtaccagcg gtcaattgta gtggaggtgc tagcgcatgc ggaattggat 3361 atcggccggc cacgcgatcg ctgacgtcgg taccctcgag tctggtaaag aaaccgctgc 3421 tgcgaaattt gaacgccagc acatggactc gtctactagc gcagcttaat taacctaggc 3481 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag 3541 gggttttttg ctgaaacctc aggcatttga gaagcacacg gtcacactgc ttccggtagt 3601 caataaaccg gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc 3661 gacgaccggg tcatcgtggc cggatcttgc ggcccctcgg cttgaacgaa ttgttagaca 3721 ttatttgccg actaccttgg tgatctcgcc tttcacgtag tggacaaatt cttccaactg 3781 atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga taagcctgtc tagcttcaag 3841 tatgacgggc tgatactggg ccggcaggcg ctccattgcc cagtcggcag cgacatcctt 3901 cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt 3961 tcgctcatcg ccagcccagt cgggcggcga gttccatagc gttaaggttt catttagcgc 4021 ctcaaataga tcctgttcag gaaccggatc aaagagttcc tccgccgctg gacctaccaa 4081 ggcaacgcta tgttctcttg cttttgtcag caagatagcc agatcaatgt cgatcgtggc 4141 tggctcgaag atacctgcaa gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg 4201 cttagctgga taacgccacg gaatgatgtc gtcgtgcaca acaatggtga cttctacagc 4261 gcggagaatc tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc 4321 tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc agcaaatcaa tatcactgtg 4381 tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt acggccagca acgtcggttc 4441 gagatggcgc tcgatgacgc caactacctc tgatagttga gtcgatactt cggcgatcac 4501 cgcttccctc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 4561 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagcta gctcactcgg 4621 tcgctacgct ccgggcgtga gactgcggcg ggcgctgcgg acacatacaa agttacccac 4681 agattccgtg gataagcagg ggactaacat gtgaggcaaa acagcagggc cgcgccggtg 4741 gcgtttttcc ataggctccg ccctcctgcc agagttcaca taaacagacg cttttccggt 4801 gcatctgtgg gagccgtgag gctcaaccat gaatctgaca gtacgggcga aacccgacag 4861 gacttaaaga tccccaccgt ttccggcggg tcgctccctc ttgcgctctc ctgttccgac 4921 cctgccgttt accggatacc tgttccgcct ttctccctta cgggaagtgt ggcgctttct 4981 catagctcac acactggtat ctcggctcgg tgtaggtcgt tcgctccaag ctgggctgta 5041 agcaagaact ccccgttcag cccgactgct gcgccttatc cggtaactgt tcacttgagt 5101 ccaacccgga aaagcacggt aaaacgccac tggcagcagc cattggtaac tgggagttcg 5161 cagaggattt gtttagctaa acacgcggtt gctcttgaag tgtgcgccaa agtccggcta 5221 cactggaagg acagatttgg ttgctgtgct ctgcgaaagc cagttaccac ggttaagcag 5281 ttccccaact gacttaacct tcgatcaaac cacctcccca ggtggttttt tcgtttacag 5341 ggcaaaagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctactg 5401 aaccgctcta gatttcagtg caatttatct cttcaaatgt agcacctgaa gtcagcccca 5461 tacgatataa gttgtaattc tcatgttagt catgccccgc gcccaccgga aggagctgac 5521 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact 5581 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 5641 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg 5701 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag 5761 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg 5821 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga 5881 tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct 5941 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt 6001 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat 6061 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg 6121 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc 6181 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa 6241 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca 6301 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt 6361 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat 6421 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg 6481 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa 6541 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc 6601 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat 6661 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc 6721 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc 6781 ttatgcgact cctgcattag gaaattaata cgactcacta taggggaatt gtgagcggat 6841 aacaattccc ctgtagaaat aattttgttt aactttaata aggagatata ccatggcaaa 6901 agcatttaag ggtaattaca agaatttaaa gaaggagaaa aagagtagga ggaatcttgt 6961 ggttgtcagt gatttaggtg gacagtatga atatggcttt gatgatatca aaacgcaaat 7021 tcttaactac ttcacattta aagcagtgag gactattttg aaccagctta atgaaatgga 7081 ctctccacaa tatcaatggt tcaatgatta tgttacagta aataagctta gggatgggaa 7141 gcgttttatt cgcaaccttg ctaaggagaa gcaagaactc gctgaaaaag tgatggtaac 7201 acggctcagt ctgtatgcga aatggataaa aaaatgtgat cacgaggaga tctacaaccg 7261 aatatctgat cagaatgtgg aagtgatgcg tgagcggctc atggaaactg tgatttggcc 7321 atcagatgac acgaactcgg agagagttgg gtgaggcgcg cgtaatacga ctcactatag 7381 ggagaccaca acggtttccc tctagaaata attttgttta actttaagaa ggagatataa 7441 ct //