LOCUS       sequence-298010-        4952 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Same as pLz47 but with randomized N*-peptide sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4952)
  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 4952)
  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: pLz55_polyPro_random_Npeptide
FEATURES             Location/Qualifiers
     source          1..4952
                     /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(482..500)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      496..543
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(909..914,915..931,932..937))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(964..982)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1050..1154
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1155..1223,1224..2015)
                     /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(1373..1392)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2186..2774
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2675..2694
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2928..2945
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2960..3099
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3025..3044)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3185..3207
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3201..3392)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4850..4869
                     /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 gacgccgcaa agggttgcgt gccgctggac gtgcccttca
      361 ggtagagcaa gcagatagcc gtgcaggtgg cggtagcgga ggtggtagtg gtggaagcgg
      421 ttaagcggtt aagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
      481 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
      541 ttgctgaaag gaggaactat atccggatat ccacaggacg ggtgtggtcg ccatgatcgc
      601 gtagtcgata gtggctccaa gtagcgaagc gagcaggact gggcggcggc caaagcggtc
      661 ggacagtgct ccgagaacgg gtgcgcatag aaattgcatc aacgcatata gcgctagcag
      721 cacgccatag tgactggcga tgctgtcgga atggacgata tcccgcaaga ggcccggcag
      781 taccggcata accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat
      841 gagcgcattg ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa
      901 actaccgcat taaagcttat cgatgataag ctgtcaaaca tgagaattct tgaagacgaa
      961 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     1021 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1081 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1141 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1201 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1261 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1321 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1381 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     1441 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     1501 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     1561 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     1621 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     1681 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     1741 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     1801 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     1861 gtgagcgtgg ctctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     1921 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     1981 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     2041 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     2101 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2161 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2221 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2281 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     2341 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2401 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2461 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     2521 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     2581 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     2641 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     2701 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     2761 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     2821 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     2881 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     2941 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3001 cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     3061 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     3121 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     3181 gcctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     3241 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     3301 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     3361 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     3421 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     3481 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     3541 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     3601 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     3661 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     3721 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     3781 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     3841 acgatcatgc gcacccgtgg ccaggaccca acgctgcccg agatgcgccg cgtgcggctg
     3901 ctggagatgg cggacgcgat ggatatgttc tgccaagggt tggtttgcgc attcacagtt
     3961 ctccgcaaga attgattggc tccaattctt ggagtggtga atccgttagc gaggtgccgc
     4021 cggcttccat tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac gcggggaggc
     4081 agacaaggta tagggcggcg cctacaatcc atgccaaccc gttccatgtg ctcgccgagg
     4141 cggcataaat cgccgtgacg atcagcggtc caatgatcga agttaggctg gtaagagccg
     4201 cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac ctgcctggac agcatggcct
     4261 gcaacgcggg catcccgatg ccgccggaag cgagaagaat cataatgggg aaggccatcc
     4321 agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc atgccggcga
     4381 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     4441 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     4501 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     4561 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     4621 tgactgggtt gaaggctctc aagggcatcg gtcgacgctc tcccttatgc gactcctgca
     4681 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     4741 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     4801 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     4861 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     4921 cgtagaggat cgagatctcg atcccgcgaa at
//
