LOCUS       sequence-212944-        6143 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of Lbpro .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6143)
  AUTHORS   Guarne A, Tormo J, Kirchweger R, Pfistermueller D, Fita I, Skern T
  TITLE     Structure of the foot-and-mouth disease virus leader protease: a 
            papain-like fold adapted for self-processing and eIF4G recognition.
  JOURNAL   EMBO J. 1998 Dec 15;17(24):7469-79. doi: 10.1093/emboj/17.24.7469.
  PUBMED    9857201
REFERENCE   2  (bases 1 to 6143)
  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: 
            "10.1093/emboj/17.24"; journalName: "EMBO J"; date: "1998-12-15- 
            15"; volume: "17"; issue: "24"; pages: "7469-79"
COMMENT     Sequence Label: Lbpro (29-195)
FEATURES             Location/Qualifiers
     source          1..6143
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     106..125
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     209..228
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        209..227
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    228..252
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             267..289
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(853..871)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      867..914
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1089..1094,1095..1111,1112..1117))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1144..1162)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1230..1334
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1335..1403,1404..2195)
                     /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(1553..1572)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2366..2954
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2855..2874
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3108..3125
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3140..3279
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3205..3224)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3365..3387
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3381..3572)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5927..5946
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5966..6043)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
       61 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
      121 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
      181 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg
      241 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat ataccatgga
      301 gttaacactg tacaacggtg agaagaagac cttttactcc aggcccaata accacgacaa
      361 ctgctggttg aacgccatcc tccagttgtt caggtacgtt gaagaaccat tcttcgactg
      421 ggtctacagt tcgcctgaga acctcacgct tgaagccatc aagcagttgg aggatctcac
      481 aggacttgaa ctgcatgagg gtggaccacc tgctctcgtg atctggaaca tcaagcactt
      541 gctccacacc ggcatcggca ccgcctcgcg acccagcgag gtgtgcgtgg tggatggtac
      601 ggacatgtgc ttggctgatt tccatgctgg cattttcctt aaggggcaag aacacgctgt
      661 gtttgcgtgt gtcacctcca acgggtggta cgcgattgac gatgaggact tctacccctg
      721 gacgccggac ccgtccgacg ttctggtgtt tgtcccgtac gatcaagaac cactcaacgg
      781 ggaatggaaa gccaagtgat aaggatccgg ctgctaacaa agcccgaaag gaagctgagt
      841 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
      901 tgaggggttt tttgctgaaa ggaggaacta tatccggata tcccgcaaga ggcccggcag
      961 taccggcata accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat
     1021 gagcgcattg ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa
     1081 actaccgcat taaagcttat cgatgataag ctgtcaaaca tgagaattct tgaagacgaa
     1141 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     1201 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1261 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1321 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1381 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1441 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1501 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1561 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     1621 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     1681 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     1741 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     1801 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     1861 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     1921 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     1981 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     2041 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     2101 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     2161 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     2221 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     2281 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2341 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2401 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2461 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     2521 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2581 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2641 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     2701 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     2761 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     2821 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     2881 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     2941 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3001 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3061 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3121 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3181 cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     3241 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     3301 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     3361 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     3421 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     3481 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     3541 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     3601 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     3661 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     3721 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     3781 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     3841 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     3901 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     3961 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     4021 acgatcatgc gcacccgtgg ccaggaccca acgctgcccg agatgcgccg cgtgcggctg
     4081 ctggagatgg cggacgcgat ggatatgttc tgccaagggt tggtttgcgc attcacagtt
     4141 ctccgcaaga attgattggc tccaattctt ggagtggtga atccgttagc gaggtgccgc
     4201 cggcttccat tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac gcggggaggc
     4261 agacaaggta tagggcggcg cctacaatcc atgccaaccc gttccatgtg ctcgccgagg
     4321 cggcataaat cgccgtgacg atcagcggtc caatgatcga agttaggctg gtaagagccg
     4381 cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac ctgcctggac agcatggcct
     4441 gcaacgcggg catcccgatg ccgccggaag cgagaagaat cataatgggg aaggccatcc
     4501 agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc atgccggcga
     4561 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     4621 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     4681 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     4741 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     4801 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
     4861 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac tgtcgtgcca
     4921 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg
     4981 tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct
     5041 gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga
     5101 tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg
     5161 agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca
     5221 tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg
     5281 tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt
     5341 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta
     5401 atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca
     5461 gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat
     5521 caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat
     5581 ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg
     5641 ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt
     5701 gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg
     5761 aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg
     5821 gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt
     5881 ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga
     5941 catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct
     6001 atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct
     6061 cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc
     6121 gccgccgcaa ggaatggtgc atg
//
