LOCUS       sequence-298029-        4949 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Same as pLz47 but with 20 polar amino acids instead of N*- peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4949)
  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 4949)
  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: pLz54_polyPro_polar
FEATURES             Location/Qualifiers
     source          1..4949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(294..312)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      308..355
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(721..726,727..743,744..749))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(776..794)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        862..966
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(967..1035,1036..1827)
                     /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(1185..1204)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1998..2586
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2487..2506
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2740..2757
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2772..2911
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2837..2856)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2997..3019
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3013..3204)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4662..4681
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4765..4784
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4765..4783
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4815..4837
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4856..4873
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4874..4894
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
ORIGIN
        1 cgccaccgcc agctcctgca cctgcgcctg ctcctccgcc accccctcca ccgccacctg
       61 ctcctgctcc tgctccggca ccaccacctc cgcctccgcc acctggaggt ggatcaggag
      121 gtggttcagg tggaggaggg acagggtcca caggatccgg atcgtcgtct tccggctccg
      181 gaacatctgg gacaggcggt ggcggtagcg gaggtggtag tggtggaagc ggttaagcgg
      241 ttaagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
      301 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
      361 aggaggaact atatccggat atccacagga cgggtgtggt cgccatgatc gcgtagtcga
      421 tagtggctcc aagtagcgaa gcgagcagga ctgggcggcg gccaaagcgg tcggacagtg
      481 ctccgagaac gggtgcgcat agaaattgca tcaacgcata tagcgctagc agcacgccat
      541 agtgactggc gatgctgtcg gaatggacga tatcccgcaa gaggcccggc agtaccggca
      601 taaccaagcc tatgcctaca gcatccaggg tgacggtgcc gaggatgacg atgagcgcat
      661 tgttagattt catacacggt gcctgactgc gttagcaatt taactgtgat aaactaccgc
      721 attaaagctt atcgatgata agctgtcaaa catgagaatt cttgaagacg aaagggcctc
      781 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
      841 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
      901 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
      961 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     1021 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     1081 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     1141 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     1201 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     1261 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     1321 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     1381 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     1441 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     1501 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     1561 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     1621 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     1681 ggctctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     1741 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     1801 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     1861 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     1921 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1981 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2041 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2101 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     2161 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2221 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2281 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2341 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2401 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2461 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2521 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2581 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2641 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2701 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2761 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     2821 aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc
     2881 gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc
     2941 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgcctccgg
     3001 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta
     3061 aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag
     3121 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag
     3181 ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg
     3241 ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca
     3301 tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca
     3361 gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca
     3421 gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt
     3481 ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt
     3541 cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg
     3601 ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat
     3661 gcgcacccgt ggccaggacc caacgctgcc cgagatgcgc cgcgtgcggc tgctggagat
     3721 ggcggacgcg atggatatgt tctgccaagg gttggtttgc gcattcacag ttctccgcaa
     3781 gaattgattg gctccaattc ttggagtggt gaatccgtta gcgaggtgcc gccggcttcc
     3841 attcaggtcg aggtggcccg gctccatgca ccgcgacgca acgcggggag gcagacaagg
     3901 tatagggcgg cgcctacaat ccatgccaac ccgttccatg tgctcgccga ggcggcataa
     3961 atcgccgtga cgatcagcgg tccaatgatc gaagttaggc tggtaagagc cgcgagcgat
     4021 ccttgaagct gtccctgatg gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg
     4081 ggcatcccga tgccgccgga agcgagaaga atcataatgg ggaaggccat ccagcctcgc
     4141 gtcgcgaacg ccagcaagac gtagcccagc gcgtcggccg ccatgccggc gataatggcc
     4201 tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc
     4261 aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc
     4321 tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag
     4381 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg
     4441 ttgaaggctc tcaagggcat cggtcgacgc tctcccttat gcgactcctg cattaggaag
     4501 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag
     4561 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     4621 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     4681 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     4741 atcgagatct cgatcccgcg aaattaatac gactcactat agggacacca caacggtttc
     4801 cctctagaaa taattttgtt taactttaag aaggagatat accatgtcct ggaagcacca
     4861 tcaccatcac catgaaaacc tgtacttcca gtccgcagct gttggtgggg gatctggagg
     4921 tgggtctggt ggacctcccc cgcctccac
//
