LOCUS       sequence-449981-        5700 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 5700)
  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 5700)
  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: pJB65
FEATURES             Location/Qualifiers
     source          1..5700
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     132..149
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    164..303
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(229..248)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     389..411
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(405..596)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2054..2073
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2157..2176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2157..2175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2207..2229
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2242..2259
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3301..3324
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(3386..3404)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3400..3447
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3813..3818,3819..3835,3836..3841))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3868..3886)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3954..4058
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4059..4127,4128..4919)
                     /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(4277..4296)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5090..5678
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5579..5598
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
       61 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
      121 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
      181 tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc
      241 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct
      301 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
      361 tccgcttaca gacaagctgt gaccgcctcc gggagctgca tgtgtcagag gttttcaccg
      421 tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat
      481 tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat
      541 gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat
      601 gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg
      661 atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt
      721 aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag
      781 cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag
      841 atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg
      901 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
      961 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
     1021 agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggccagga cccaacgctg
     1081 cccgagatgc gccgcgtgcg gctgctggag atggcggacg cgatggatat gttctgccaa
     1141 gggttggttt gcgcattcac agttctccgc aagaattgat tggctccaat tcttggagtg
     1201 gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc cggctccatg
     1261 caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgcctaca atccatgcca
     1321 acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc ggtccaatga
     1381 tcgaagttag gctggtaaga gccgcgagcg atccttgaag ctgtccctga tggtcgtcat
     1441 ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa
     1501 gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag acgtagccca
     1561 gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
     1621 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
     1681 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
     1741 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
     1801 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgac
     1861 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     1921 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     1981 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     2041 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     2101 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat
     2161 acgactcact atagggacac cacaacggtt tccctctaga aataattttg tttaacttta
     2221 agaaggagat ataccatgtc ccaccatcat catcatcatg cgccaaaaaa ggttgaaacc
     2281 ggtcacaccc tatccggcgg atctgcagct gcaacgcaag tggatcgtaa atctcatgct
     2341 gcagctaacg agccgggcac ttgccgagca gcagctgcta ccgagtacgt aggtactgag
     2401 caattcacct ctttttgcaa taccagcccc aagccaaatg cgacgaaggt caatgtgacc
     2461 acaacggctc gtggtcgccc tgctgcagct acggaagttt cacggaccga gaaggcagca
     2521 gctaacgaat ccggtgtctg ccgtaacgct gctgcaaccg aatacatgag taatgaagct
     2581 cacttttctc tatgtggaac agccgcaaag ccttcacaag cggataaagt catgttcggc
     2641 gccacagcac gaacgcatca agtggcagct gctagtgatg aattcaggcc ctcttctgct
     2701 gcagcaaacg aatcgggtgc aaaacgcaca attaccggct cgcagtacgc agacgaaggt
     2761 cttgcgcgac tcacgatcaa cggagcacct gcaaaagtag ccagaaccca cacctttgcg
     2821 ggctctgacg cagctgcaac ggaaatcggt cgctctactc gcgcagcagc tgatgaaagc
     2881 ggttcgtgtc gttcaatctc aggcaccgag tatctcagta acgagcaatt ccaatccttt
     2941 tgtgacacaa aacctcaacg cagcccgttc aaggttggcc aagatcgcac gaacaagggt
     3001 cagtctgcag ctgcaaactt ggttgatcgt tccgaactgg ctgcagctaa cgaaccaggt
     3061 tcatgctctc gggctgctgc atctcagtat ggccaaagca aaatctgcgg tggtggcgtg
     3121 ggaaaagtgc gctcaatgcg cacccttcgc ggcacctcag tatctggcca acagctagat
     3181 catgcaccta agatgtccgg tgacgagcgc ggcgggtgca tgcccgtcac cggtaatgag
     3241 tactacggtc gtgaacattt cgaaccgttt tgtacgagca ccccagagcc cgaagctcaa
     3301 tggagtcatc cacagttcga aaagtaaagc ggttaagatc cggctgctaa caaagcccga
     3361 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
     3421 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg atatccacag
     3481 gacgggtgtg gtcgccatga tcgcgtagtc gatagtggct ccaagtagcg aagcgagcag
     3541 gactgggcgg cggccaaagc ggtcggacag tgctccgaga acgggtgcgc atagaaattg
     3601 catcaacgca tatagcgcta gcagcacgcc atagtgactg gcgatgctgt cggaatggac
     3661 gatatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta cagcatccag
     3721 ggtgacggtg ccgaggatga cgatgagcgc attgttagat ttcatacacg gtgcctgact
     3781 gcgttagcaa tttaactgtg ataaactacc gcattaaagc ttatcgatga taagctgtca
     3841 aacatgagaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     3901 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3961 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4021 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4081 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4141 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4201 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4261 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     4321 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4381 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4441 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4501 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     4561 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt
     4621 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4681 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4741 ttattgctga taaatctgga gccggtgagc gtggctctcg cggtatcatt gcagcactgg
     4801 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     4861 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4921 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4981 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5041 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5101 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5161 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5221 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5281 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5341 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5401 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5461 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5521 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5581 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5641 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
//
