LOCUS       sequence-449990-        4952 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli protein expression for protein purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4952)
  AUTHORS   Turnsek JB, Oltrogge LM, Savage DF
  TITLE     Conserved and repetitive motifs in an intrinsically disordered 
            protein drive ⍺-carboxysome assembly.
  JOURNAL   J Biol Chem. 2024 Jul 4;300(8):107532. doi: 
            10.1016/j.jbc.2024.107532.
  PUBMED    38971311
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"; journalName: "J Biol 
            Chem."; date: "2024-07-4"; volume: "300(8):107532. doi"; pages: " 
            10.1016/j.jbc.2024.107532"
COMMENT     Sequence Label: pLz48
FEATURES             Location/Qualifiers
     source          1..4952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..192)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1650..1669
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1753..1772
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1753..1771
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1803..1825
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1838..1861
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1862..1882
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(2234..2252)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2248..2295
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2661..2666,2667..2683,2684..2689))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2716..2734)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2802..2906
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2907..2975,2976..3767)
                     /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(3125..3144)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3938..4526
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4427..4446
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4680..4697
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4712..4851
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4777..4796)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4937..4952,1..7)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
       61 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
      121 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
      181 ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc
      241 gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact
      301 ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac
      361 tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca
      421 gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag
      481 actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt
      541 gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag
      601 gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg
      661 ccaggaccca acgctgcccg agatgcgccg cgtgcggctg ctggagatgg cggacgcgat
      721 ggatatgttc tgccaagggt tggtttgcgc attcacagtt ctccgcaaga attgattggc
      781 tccaattctt ggagtggtga atccgttagc gaggtgccgc cggcttccat tcaggtcgag
      841 gtggcccggc tccatgcacc gcgacgcaac gcggggaggc agacaaggta tagggcggcg
      901 cctacaatcc atgccaaccc gttccatgtg ctcgccgagg cggcataaat cgccgtgacg
      961 atcagcggtc caatgatcga agttaggctg gtaagagccg cgagcgatcc ttgaagctgt
     1021 ccctgatggt cgtcatctac ctgcctggac agcatggcct gcaacgcggg catcccgatg
     1081 ccgccggaag cgagaagaat cataatgggg aaggccatcc agcctcgcgt cgcgaacgcc
     1141 agcaagacgt agcccagcgc gtcggccgcc atgccggcga taatggcctg cttctcgccg
     1201 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat
     1261 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg
     1321 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
     1381 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     1441 aagggcatcg gtcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
     1501 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc
     1561 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
     1621 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     1681 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg
     1741 atcccgcgaa attaatacga ctcactatag ggacaccaca acggtttccc tctagaaata
     1801 attttgttta actttaagaa ggagatatac catgtcctgg agccacccgc agttcgaaaa
     1861 ggaaaacctg tacttccagt ccgctgatgt aactggtatt gctctgggta tgatcgaaac
     1921 acgtggctta gttcctgcga ttgaagcagc ggacgccatg actaaagcgg ctgaagtgcg
     1981 tttggtcggt cgtcaatttg ttggtggcgg ttacgtcacc gtattggttc ggggcgaaac
     2041 aggcgctgtt aacgccgctg ttcgtgctgg cgccgatgct tgcgaacgtg ttggtgatgg
     2101 cttggttgct gcgcacatca ttgcgcgtgt ccactcagaa gtagaaaaca ttctgcctaa
     2161 ggcgccacaa gcctaagcgg ttaagatccg gctgctaaca aagcccgaaa ggaagctgag
     2221 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     2281 ttgaggggtt ttttgctgaa aggaggaact atatccggat atccacagga cgggtgtggt
     2341 cgccatgatc gcgtagtcga tagtggctcc aagtagcgaa gcgagcagga ctgggcggcg
     2401 gccaaagcgg tcggacagtg ctccgagaac gggtgcgcat agaaattgca tcaacgcata
     2461 tagcgctagc agcacgccat agtgactggc gatgctgtcg gaatggacga tatcccgcaa
     2521 gaggcccggc agtaccggca taaccaagcc tatgcctaca gcatccaggg tgacggtgcc
     2581 gaggatgacg atgagcgcat tgttagattt catacacggt gcctgactgc gttagcaatt
     2641 taactgtgat aaactaccgc attaaagctt atcgatgata agctgtcaaa catgagaatt
     2701 cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     2761 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     2821 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2881 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     2941 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3001 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3061 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3121 ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc
     3181 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     3241 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3301 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     3361 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3421 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     3481 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     3541 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3601 aatctggagc cggtgagcgt ggctctcgcg gtatcattgc agcactgggg ccagatggta
     3661 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3721 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     3781 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     3841 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     3901 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3961 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4021 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4081 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4141 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4201 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4261 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4321 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4381 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4441 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4501 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4561 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     4621 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     4681 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     4741 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata
     4801 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     4861 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4921 caagctgtga ccgcctccgg gagctgcatg tg
//
