LOCUS       sequence-298009-        5039 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Same as pLz47 but with MR repeat 1 instead of N*-peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5039)
  AUTHORS   Oltrogge LM, Chaijarasphong T, Chen AW, Bolin ER, Marqusee S, Savage
            DF
  TITLE     Multivalent interactions between CsoS2 and Rubisco mediate 
            alpha-carboxysome formation.
  JOURNAL   Nat Struct Mol Biol. 2020 Mar;27(3):281-287. doi: 
            10.1038/s41594-020-0387-7. Epub 2020 Mar 2.
  PUBMED    32123388
REFERENCE   2  (bases 1 to 5039)
  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/s41594-020-0387-7"; journalName: "Nat Struct Mol Biol"; 
            date: "2020-03"; volume: "27"; issue: "3"; pages: "281-287"
COMMENT     Sequence Label: pLz56_polyPro_CsoS2_MR_r1
FEATURES             Location/Qualifiers
     source          1..5039
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             92..109
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             110..130
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(569..587)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      583..630
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(996..1001,1002..1018,1019..1024))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1051..1069)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1137..1241
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1242..1310,1311..2102)
                     /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(1460..1479)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2273..2861
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2762..2781
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3015..3032
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3047..3186
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3112..3131)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3272..3294
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3288..3479)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4937..4956
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg acaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacca tgtcctggaa gcaccatcac catcaccatg aaaacctgta
      121 cttccagtcc gcagctgttg gtgggggatc tggaggtggg tctggtggac ctcccccgcc
      181 tccaccgcca ccgccagctc ctgcacctgc gcctgctcct ccgccacccc ctccaccgcc
      241 acctgctcct gctcctgctc cggcaccacc acctccgcct ccgccacctg gaggtggatc
      301 aggaggtggt tcaggtggag gaaaggttga aaccggtcac accctatccg gcggatctgt
      361 tacgggcacg caagtggatc gtaaatctca tgtgactgga aacgagccgg gcacttgccg
      421 agcagtcacg ggcaccgagt acgtaggtac tgagcaattc acctcttttt gcggtggcgg
      481 tagcggaggt ggtagtggtg gaagcggtta agcggttaag atccggctgc taacaaagcc
      541 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
      601 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatatcca
      661 caggacgggt gtggtcgcca tgatcgcgta gtcgatagtg gctccaagta gcgaagcgag
      721 caggactggg cggcggccaa agcggtcgga cagtgctccg agaacgggtg cgcatagaaa
      781 ttgcatcaac gcatatagcg ctagcagcac gccatagtga ctggcgatgc tgtcggaatg
      841 gacgatatcc cgcaagaggc ccggcagtac cggcataacc aagcctatgc ctacagcatc
      901 cagggtgacg gtgccgagga tgacgatgag cgcattgtta gatttcatac acggtgcctg
      961 actgcgttag caatttaact gtgataaact accgcattaa agcttatcga tgataagctg
     1021 tcaaacatga gaattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
     1081 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     1141 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     1201 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     1261 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     1321 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     1381 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     1441 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     1501 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     1561 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     1621 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     1681 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     1741 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
     1801 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     1861 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     1921 ggtttattgc tgataaatct ggagccggtg agcgtggctc tcgcggtatc attgcagcac
     1981 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     2041 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     2101 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     2161 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     2221 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     2281 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     2341 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     2401 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     2461 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     2521 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     2581 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     2641 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     2701 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     2761 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     2821 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     2881 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     2941 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     3001 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     3061 ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg
     3121 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     3181 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     3241 gcatccgctt acagacaagc tgtgaccgcc tccgggagct gcatgtgtca gaggttttca
     3301 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
     3361 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
     3421 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
     3481 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag
     3541 aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag
     3601 ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc
     3661 cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg
     3721 cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca
     3781 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg
     3841 cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag
     3901 cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggcca ggacccaacg
     3961 ctgcccgaga tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga tatgttctgc
     4021 caagggttgg tttgcgcatt cacagttctc cgcaagaatt gattggctcc aattcttgga
     4081 gtggtgaatc cgttagcgag gtgccgccgg cttccattca ggtcgaggtg gcccggctcc
     4141 atgcaccgcg acgcaacgcg gggaggcaga caaggtatag ggcggcgcct acaatccatg
     4201 ccaacccgtt ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc agcggtccaa
     4261 tgatcgaagt taggctggta agagccgcga gcgatccttg aagctgtccc tgatggtcgt
     4321 catctacctg cctggacagc atggcctgca acgcgggcat cccgatgccg ccggaagcga
     4381 gaagaatcat aatggggaag gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc
     4441 ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg
     4501 cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca
     4561 ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg
     4621 ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag
     4681 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
     4741 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     4801 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     4861 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     4921 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     4981 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaat
//
