LOCUS       sequence-434504-        2315 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  8xHis-tag fused to PreScission Protease cleavage site for protein 
            purification and cleavage.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2315)
  AUTHORS   Hieronimus K, Donauer T, Klein J, Hinkel B, Spanle JV, Probst A, 
            Niemeyer J, Kibrom S, Kiefer AM, Schneider L, Husemann B, Bischoff 
            E, Mohring S, Bayer N, Klein D, Engels A, Ziehmer BG, Stiebeta J, 
            Moroka P, Schroda M, Deponte M
  TITLE     A Modular Cloning Toolkit for the production of recombinant proteins
            in Leishmania tarentolae.
  JOURNAL   Microb Cell. 2024 Apr 30;11:128-142. doi: 10.15698/mic2024.04.821. 
            eCollection 2024.
  PUBMED    38799406
REFERENCE   2  (bases 1 to 2315)
  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: "Microb 
            Cell."; date: "2024-04-30"; volume: "11:128-142. doi"; pages: " 
            10.15698/mic2024.04.821. eCollection 2024"
COMMENT     Sequence Label: pMBS-849
FEATURES             Location/Qualifiers
     source          1..2315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     139..156
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             237..260
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             276..299
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(338..356)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        424..528
                     /gene="bla"
                     /label=AmpR promoter
     CDS             750..1541
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1712..2300
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2201..2220
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
       61 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
      121 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
      181 aaaccgcctc tccccgcgcg ttggccgatt cattaatcac tctgtggtct caaatgcacc
      241 atcaccacca tcatcaccat ggcagcggca gcggcctgga ggtgctgttc cagggcccag
      301 gttgagacca cgaagtggct cttcagtgga cgaaagggcc tcgtgatacg cctattttta
      361 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
      421 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
      481 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gcgctcacgc
      541 aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg
      601 gcttgttatg actgtttttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag
      661 cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg
      721 gcagtcgccc taaaacaaag ttaaacatca tgagggaagc ggtgatcgcc gaagtatcga
      781 ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg
      841 tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagcgat attgatttgc
      901 tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt
      961 tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg
     1021 ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag
     1081 aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc
     1141 tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg
     1201 aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa
     1261 cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt
     1321 cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact
     1381 gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt
     1441 atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc
     1501 attacgtaaa aggcgagatc accaaggtag tcggcaaata actgtcagac caagtttact
     1561 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1621 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1681 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1741 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1801 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1861 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1921 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1981 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2041 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2101 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2161 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2221 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2281 gggggcggag cctatggaaa aacgccagca acgcg
//
