LOCUS       sequence-472471-       10372 bp    DNA     circular SYN 13-MAY-2025
DEFINITION  expresses A. thaliana Cpn60-alpha Cpn60-beta Cpn20 Raf1 Raf2 RbcX 
            Bsd2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10372)
  TITLE     Effects of chaperone selectivity on the assembly of plant Rubisco 
            orthologs in E. coli
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10372)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pTW334e
FEATURES             Location/Qualifiers
     source          1..10372
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             86..877
                     /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        join(1194..1199,1200..1215,1216..1222)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1230..1246
                     /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             4507..4529
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             5796..5804
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     RBS             6222..6244
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    7966..7982
                     /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        complement(join(7990..7996,7997..8012,8013..8018))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     promoter        8261..8338
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             8339..9421
                     /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     complement(8358..8377)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      9578..10316
                     /direction=RIGHT
                     /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."
     promoter        join(10366..10372,1..85)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
       61 tcaataatat tgaaaaagga agagtatgag ggaagcggtg atcgccgaag tatcgactca
      121 actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca
      181 tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt
      241 tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga
      301 aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt
      361 gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg
      421 gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc
      481 tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga
      541 actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct
      601 atggaactcg ccgcccgact gggctggaga tgagcgaaat gtagtgctta cgttgtcccg
      661 catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc
      721 aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct
      781 tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta
      841 cgtgaaaggc gagatcacca aggtagtcgg caaataaacg ccatgggcat taaaaaatgc
      901 cctcttgggt tatcaagagg gtcattatat ttgctgagga gtgccgttaa tgcgcaacgc
      961 aattaatgta agttagctca ctcattaggc acaattctca tgtttgacag cttatcatcg
     1021 actgcacggt gcaccaatgc ttctggcgtc aggcagccat cggaagctgt ggtatggctg
     1081 tgcaggtcgt aaatcactgc ataattcgtg tcgctcaagg cgcactcccg ttctggataa
     1141 tgttttttgc gccgacatca taacggttct ggcaaatatt ctgaaatgag ctgttgacaa
     1201 ttaatcatcg gctcgtataa tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     1261 cagccagtcc gtttaggtgt tttcacgagc acttcaccaa caacgcgtac aactaggaga
     1321 tataccatgg gtatgcaaca gctctaccaa ccgttccgac caccgtcatc tcccattcca
     1381 acccaattcc gttcactcga ctccgccggt aaaatcgaaa tcctcgccgg tcgaatggct
     1441 ctctggttcg aatacgcacc tcttatctct tctctttaca ccgatggttt cactcctcca
     1501 accatcgaag aactcaccgg aatctcaagc atcgaacaga accgtttaat cgtcggtgcg
     1561 caagttcgtg actcaattct tcaatctatc catgaaccgg aacttatttc tgcgttcgac
     1621 accggtggtg cagagcttct atacgaaatt cgtctcctta gcaccacgca gcgtgttgca
     1681 gccgccacat tcatcattga tcgtaatatt gattcgaaag gagcacagga tttggcacga
     1741 gccatcaagg attatcctaa ccgtcgtgga gacgttggat ggttggattt tgattacaac
     1801 ttaccaggag attgtctttc attcttgtat tacaggcaga gcagagagaa taagaatcca
     1861 tcggatcaga gaacctcgat gcttctacaa gcattggggg ttgcagaatc agaaaaggct
     1921 aagaacaggt tgaacacaga gctatacggg gacaaggaag cagagaagga gaaggagaag
     1981 aagaagaagg aagaggaagt caaagctatt cggattccag tggtgaggct taagtttgga
     2041 gaagtagcag aagcgacctc ggtggtagtt ttaccggttt gtaaagcaga ggaaggggaa
     2101 aagaagattc ttgaagctcc aatggagatc atagccggag gggattttaa ggtggttgag
     2161 gcggagaaag gatggaagag atgggtagtg cttccgtcgt ggaacccagt ggcagccatt
     2221 gggaaaggcg gtgtggcggt ttctttcagg gatgatagaa aagtgctgcc ttgggatgga
     2281 aaggaggagc ctttactggt agtggccgat agggtgagga atgttgtgga ggctgatgac
     2341 gggtattatc tcgtggtggc tgagaacgga cttaagctag agaaaggatc agatttgaag
     2401 gcgagagagg tgaaggagag tttagggatg gttgttttgg tggtgaggcc gccaagagaa
     2461 gatgatgatg attggcagac aagtcatcag aactgggact gaattacaaa aattaggaga
     2521 tatacatatg atgtctaatc tggcgcagga ttttcttggt gacttcggag ctcgtgaccc
     2581 ttacccggag gagatagcga gtcaattcgg agataaagtg ttgggatgcc aaagcactga
     2641 gcacaagatt ttgataccaa acgcatctgt tttgtctctc tcccagcttc agtgttcccc
     2701 tgtttcgtct tcacagcctc ctttgtccgg cgatgatgcc agaactctcc tccacaaggt
     2761 tttgggatgg agtatagtgg ataatgaagc gggtggtctg aaaataaggt gtatgtggaa
     2821 agtgagggat tttgggtgcg gtgttgaact cataaacagg atccataagg ttgctgaagc
     2881 ttctggtcat tacccttctc ttcacttgga aagtcctacc caagttcgag ctgaactatt
     2941 tacctcttct atcggagggt tgagcatgaa tgatttcata atggcggcta aaatagatga
     3001 tatcaagact tctgatcttt cccctaggaa aagagcttgg gcgtgacggc tgaaattaat
     3061 gaggagatat acatatggta tcgagaattg cgaatcgaaa gagcaaaaag ctcttgattg
     3121 tgaatgagga tgttgctggt aattacgacg ataccttcgg tgatgtacaa aagcaaattg
     3181 ttaattattt tacatacaaa gctgtgagga cggttcttca tcagctgtac gaaatgaatc
     3241 ctcctcaata cacttggttc tataatcata tcataacaaa cagaccgacg gatggcaaac
     3301 gtttcctccg tgccctcggc aaggagagtc aggagcttgc agaaagagtg atgatcacgc
     3361 gtcttcactt gtatggcaaa tggatcaaga aatgcgacca tgggaagata taccaggaaa
     3421 tatcggatga aaacttggcg ttgatgcgtg aacgcctgat ggagacagtg atatggcctt
     3481 ccgatgacac aaactcaagg taattcagct aagacactgc acaggagata tacatatgat
     3541 ggccgcaaac aataatcctc agggcactaa accaaacagt ttagtttgtg caaattgtga
     3601 aggggaaggt tgtgtggcat gttctcaatg caaaggaggt ggtgtgaatt tgattgatca
     3661 tttcaatggt cagtttaaag ctggtgcact ctgttggctt tgcagaggta aaaaggaggt
     3721 gttgtgtggg gactgcaatg gagctggttt cattggtggg ttcttgagca ccttcgatga
     3781 gtagagtcag tagggcccta aaaaaaaacc ccgcccctga cagggcaggg ttttttttgg
     3841 cgaaaaaacc ccgctgaagc ggggtttttt gcgtacattg gctccgtcta agaaagtata
     3901 gccatcacat ctgaagctct gagggcaatg tagttggaac catctttgcc cttgaagtcg
     3961 ttaccagcat acttggagta aagtactgtg cttccggttg atactggtag aggcgtaatt
     4021 ttaccttcct cgtctaggga acccggtcca actgctatca cggtgccaat agaaggcttc
     4081 tctttggtag tctcggttaa caacaaccct ccagctgttt tctcctccgc ctcagcaacc
     4141 ttaataaaga ctcggtcatt caaaggtttg agatctttga tgtcctctgt ctcaagaatg
     4201 ccaacaatat catcttcctt gaggataaga tgcttcacat cattgaactc cacctcagtt
     4261 cctgcgtatt tggagtagat aatttgtgct ccagtaggga cagtgatatc aattttgttc
     4321 ttcccaatag ttcttccttc acccacagca acgacttcac ctccttgagg ttttgattga
     4381 gcagtggatg gaagtaagat accacctaaa gtcttctcct ctgcctcctt gatcttcacc
     4441 aaaactcgat ctcccaatgg cttaattgaa gtatacttag gggcaacaac agaggccata
     4501 tgtatatctc cttcttaaag ttaaacaaaa ttatttctag agggcaattg ttatccggtc
     4561 gacgtcgacc ttagtatccg taacctgagt tgtccattgg gttgccgacg ggaactggtt
     4621 caggttcctt gatctcaaca acaacgcagt ccgacatcaa gaaagtcttt gcaaccgaag
     4681 ctgcgtgttc caagcaacat ctcacaacct ttgttggatc gatgattcct gcagccatca
     4741 aatcctcgta cttgccggtt gcagcattgt aaccgaattt cacgttatcg ttggaaagca
     4801 ccttctcgct aactacactt ccattaacac cagcattctt ggcgattagt tttagggggt
     4861 aacttagtgc tcttttaact atatccgctc caaccttttc ttcatcatta tcaagcgtgg
     4921 ctttaatggc atcaaccttg gaagcaagac gaagcagagt gcaaccacca ccaacaacaa
     4981 taccttcctc gacagcggcc tttgttgcat taagagcatc ttcaactctc aacttcttct
     5041 ctttgagttc tgtctcagtt tgtgctccaa cctgaatcac agcaactcca ccagagagct
     5101 ttgcaattct ctcattgagt ttctccttct cgtaatcttg ctccgcttgc tcaataagat
     5161 ttttaatctg tgtaacacgc tttttcactg catcctgtgt gcttccatca cccactatgg
     5221 tcgaggtttc ctttgtgagg acaacctttg aagcattacc aagaacctct tttccagctt
     5281 tgtcaagtga aagaccaact tcctcacgga tcacagttgc tccagttaga atggcaatat
     5341 catcaaggta ttggctcttg cgctctccaa accctggagc tctgagagct gctatcttca
     5401 gtgtacctct aagcttgttt acaacaagag tcgctaaagc ttcttgttca atgtcttcag
     5461 caataattaa tattgggtat ccgcctctaa ttgcatcctc caaaacacca acaagatccc
     5521 ttgcattggt aattttcttg tcaacgagaa gtaacttgca attgtcgaac tcgacggaca
     5581 ttttctcgct gtctgtcaca aagtaaggag agatataacc acggtcaaat tgcattcctt
     5641 ctacgacgta gaggttgttc tcggcacttt taccctcctc aagggtaacc acacctttcc
     5701 tgcccacttt gctcattgct tcagcaatca tatttccaat ttcgtcgttg ttacccgcac
     5761 taacagctgc aacatctgca agttcactgt cttcaacctc cttagacatt ttcttgagct
     5821 cggtaaccaa agcctttgct gtcttctcaa tacctctggt gatcaaaaca gggtttgcac
     5881 cagcagctac caccttgaca ccctcagcga taaaaccttg tgcaagaaca acagatgttg
     5941 ttgtaccgtc accagccaag tcattggttt tggcagccgc ttgcctcaca agcttagcgc
     6001 caatgttctc aaccgggtct tccaactcaa cctctcttgc aacagtcaca ccatcattaa
     6061 caattcgtgg tgatccatac ttgctctcaa gaacaacatt tcgtcctttg ggtccaagtg
     6121 taacgccaac aagatctgcg agcttgttga caccagcttg aagtctccta atggtagtcc
     6181 catctttgtt gaaatgtaat tcctttgctg ccatatgtat atctccttct taaagttaaa
     6241 caaaattatt tctagctcct tacaccatga ggccctcagg agcagcagca gcaggagcct
     6301 tgggtttcgg tttgtcaaca acaatggcct gagtggtcag taccattcct gctacggaag
     6361 cagcgttctg tagcgcacat cttgtcactt tagccggatc aatcactcca gcttcgaaca
     6421 gattctcgta agtatcagtc attgcgttgt acccattctc ccaatcgctg aacataatct
     6481 tttccacaac aacttctcct tcaactccag cgttctgagc aataagagca gctggtgaca
     6541 gcaatgcctt ttgtactatg tcagctccca aacgttcatc agcatcctca aaagtctcct
     6601 taatggcggg aataacagtg gagagatgca ccaaagcagc accaccacct ggaactatgc
     6661 cttcctcgat agcagcaaat gttgcgttct ttgcatcctc gatacgaagc ttacggtcct
     6721 caagctcagt ttcagttgct gctccgactt taatgacagc aacacctcca gatagcttcg
     6781 caattctctc agcgagcttc tctgagtcat acacagaatc agtctcaaat agttctttct
     6841 tcagctgagc aatccgagct tgtaattcgt ccttggaagc tgcatctgca atgagggtag
     6901 tcgaatcttt gctaatcgtg acttttcggg caatacccaa ttgatctatg gttgcgtttt
     6961 caaccagtaa gctcatgtcc atggctaggt actcagctcc tgtcaagatt gcaatatctt
     7021 gaagcatggc ttttcttctt tctccaaatc ctggcgcttt aacggcaaca acattgagga
     7081 caccacgaag cttgttcacg acaagggtag ctaaggcttc accagtaaca tcctctgcaa
     7141 taatcaacaa cggagcacga agctgagtgg tcttctccaa aatagggatg atgtctttga
     7201 tcgcagtgat cttctgatca gtgatcaata ccctagcatt ctcaaactca gctagtagtt
     7261 tctcagggtt tgtaacaaac tgaggcgaga tgtaacctct gtcaatctcc atcccttctt
     7321 caacttcgac cgtggtttca aaagaagatg aagattcaat ggacaaaaca ccatcaggtc
     7381 caactttgtc gatggcatca gcaatcattg accctataag gtcatcattt ccagcagaga
     7441 tagaagccac agctctgatg tcatcacgac ctttcacggg cctagctttc ttctgaagct
     7501 cttcgatcag accttgaaca gtcttatcaa ttcccctctt gagtgaaacg ggattcgcac
     7561 cagaagtgac gctcaacaat ccatgtttga ttatctcccg agcaaggata gacgcagtgg
     7621 ttgtcccatc accagcagag tcattagtct tactcgcaac ctcacgaata agcgctgcac
     7681 cagcattttc catggcatta ggtaactcaa tagctctagc aatggtgact ccatcattca
     7741 caaccttagg acttccaaat tcatccaaca caacattcct ccctctgggg ccaagcgtaa
     7801 gaccaacaca atcagcaagc ttatcaatac cagcttgtag agcagctcta gagtgctggt
     7861 caaaagctat ttcctttaca ttcatatgta tatctcctac tgcacctatt gttggtgaag
     7921 tgctcgtgaa aacacctaaa cggactggct gtttcctgtg tgaaattgtt atccgctcac
     7981 aattccacac attatacgag ccgatgatta attgtcaaca gctcatttca gaatatttgc
     8041 cagaaccgtt atgatgtcgg cgcaaaaaac attatccaga acgggagtgc gccttgagcg
     8101 acacgaatta tgcagtgatt tacgacctgc acagccatac cacagcttcc gatggctgcc
     8161 tgacgccaga agcattggtg caccgtgcag tcgatgataa gctgtcaaac atgagaattg
     8221 tgcctaatga gtgagctaac ttacattaat tgcgttgcgc gacaccatcg aatggtgcaa
     8281 aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt
     8341 gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc
     8401 ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc
     8461 gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc
     8521 gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc
     8581 ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg
     8641 aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg
     8701 gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac
     8761 taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt
     8821 ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca
     8881 aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg
     8941 gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag
     9001 tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc
     9061 gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg
     9121 gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg
     9181 ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt
     9241 ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt
     9301 ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc
     9361 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     9421 aagggtccct aagtctcctc agcaaaacga aaggcccagt ctttcgactg agcctttcgt
     9481 tttatttgac cggatgtcct cttcctcctg ggctgacttc aggtgctaca tttgaagaga
     9541 taaattgcac tgaaatctag agcggttcag tagaaaagat caaaggatct tcttgagatc
     9601 ctttttttct gcgcgtaatc ttttgccctg taaacgaaaa aaccacctgg ggaggtggtt
     9661 tgatcgaagg ttaagtcagt tggggaactg cttaaccgtg gtaactggct ttcgcagagc
     9721 acagcaacca aatctgtcct tccagtgtag ccggactttg gcgcacactt caagagcaac
     9781 cgcgtgttta gctaaacaaa tcctctgcga actcccagtt accaatggct gctgccagtg
     9841 gcgttttacc gtgcttttcc gggttggact caagtgaaca gttaccggat aaggcgcagc
     9901 agtcgggctg aacggggagt tcttgcttac agcccagctt ggagcgaacg acctacaccg
     9961 agccgagata ccagtgtgtg agctatgaga aagcgccaca cttcccgtaa gggagaaagg
    10021 cggaacaggt atccggtaaa cggcagggtc ggaacaggag agcgcaagag ggagcgaccc
    10081 gccggaaacg gtggggatct ttaagtcctg tcgggtttcg cccgtactgt cagattcatg
    10141 gttgagcctc acggctccca cagatgcacc ggaaaagcgt ctgtttatgt gaactctggc
    10201 aggagggcgg agcctatgga aaaacgccac cggcgcggcc ctgctgtttt gcctcacatg
    10261 ttagtcccct gcttatccac ggaatctgtg ggtaactttg tatgtgtccg cagcgcccgc
    10321 cgcagtctca cgcccggagc gtagcgaccg agtgacggcc ttaatttggc tt
//
