LOCUS       62056_18590        7958 bp DNA     circular SYN 21-SEP-2020
DEFINITION  Shuttle-expression vector pPREx2, complete sequence.
ACCESSION   MT670350
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7958)
  AUTHORS   Bakkes PJ, Ramp P, Bida A, Dohmen-Olma D, Bott M, Freudl R.
  TITLE     Improved pEKEx2-derived expression vectors for tightly controlled 
            production of recombinant proteins in Corynebacterium glutamicum
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7958)
  AUTHORS   Bakkes PJ, Ramp P, Bida A, Dohmen-Olma D, Bott M, Freudl R.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-JUN-2020) Institute of Bio- and Geosciences; IBG-1: 
            Biotechnology, Forschungszentrum Juelich GmbH, Wilhelm-Johnen 
            Strasse, Juelich, NRW 52425, Germany
REFERENCE   3  (bases 1 to 7958)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (25-JUN-2020) Institute of Bio- and Geosciences"; volume: " IBG-1"; 
            pages: " Biotechnology, Forschungszentrum Juelich GmbH, 
            Wilhelm-Johnen Strasse, Juelich, NRW 52425, German"
FEATURES             Location/Qualifiers
     source          1..7958
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             48..71
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     primer_bind     complement(81..97)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      310..396
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      488..515
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        534..625
                     /label=AmpR promoter
     rep_origin      complement(1137..1725)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             1958..2770
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             4389..5672
                     /codon_start=1
                     /transl_table=11
                     /product="replicase"
                     /label=replicase
                     /note="Orf-1 from Corynebacterium glutamicum pBL1"
                     /protein_id="QNT09624.1"
                     /translation="MNTSKEPQVNEGSKVTRARAWRRQNVMYKITNSKALAGCHRWRRD
                     EAVAVSWSSNGASQFEGLQNSHSRWGSPLAELEVMGERRIELAIATKNHLAAGGALMMF
                     VGTVRHNRSQSFAQVEAGIKTAYSSMVKTSQWKKERARYGVEHTYSDYEVTDSWANGWH
                     LHRNMLLFLDRPLSDDELKAFEDSMFSRWSAGVVKAGMDAPLREHGVKLDQVSTWGGDA
                     AKMATYLAKGMSQELTGSATKTASKGSYTPFQMLDMLADQSDAGEDMDAVLVARWREYE
                     VGSKNLRSSWSRGAKRALGIDYIDADVRREMEEELYKLAGLEAPERVESTRVAVALVKP
                     DDWKLIQSDFAVRQYVLDCVDKAKDVAAAQRVANEVLASLGVDSTPCMIVMDDVDLDAV
                     LPTHGDATKRDLNAAVFAGNEQTILRTH"
     promoter        complement(6393..6421)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             complement(6470..7549)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(7550..7627)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     promoter        7861..7889
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    7897..7913
                     /label=lac operator
                     /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)."
ORIGIN
        1 aagcttgcat gcctgcagaa ggagatatac atatgacctg agctagctgg tcccacccac
       61 agttcgagaa gtaagaattc actggccgtc gttttacagc caagcttggc tgttttggcg
      121 gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa
      181 aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag
      241 aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact
      301 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
      361 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
      421 gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa
      481 attaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac tcttttgttt
      541 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
      601 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
      661 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
      721 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
      781 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
      841 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
      901 catacactat tctcagaatg acttggttga gtaattcgta atcatgtcat aggctcttcc
      961 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     1021 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     1081 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1141 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1201 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     1261 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     1321 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     1381 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     1441 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     1501 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     1561 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     1621 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     1681 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1741 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     1801 agattatcaa aaaggatctt cacctagatc cttttggggg gggggggaaa gccacgttgt
     1861 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     1921 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     1981 gtcttgctcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     2041 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga
     2101 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     2161 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     2221 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccgggaaaa cagcattcca
     2281 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     2341 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     2401 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     2461 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataagc ttttgccatt
     2521 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     2581 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     2641 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     2701 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     2761 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     2821 ttgacgggac ggcggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagatc
     2881 acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa aatcaccaac
     2941 tggtccacct acaacaaagc tctcatcaac cgtggctccc tcactttctg gctggatgat
     3001 ggggcgattc aggcctggta tgagtcagca acaccttctt cacgaggcag acctcagcgc
     3061 cccccccccc ctagcttgtc tacgtctgat gctttgaatc ggacggactt gccgatcttg
     3121 tatgcggtga tttttccctc gtttgcccac tttttaatgg tggccggggt gagagctacg
     3181 cgggcggcga cctgctgcgc tgtgatccaa tattcggggt cgttcactgg ttcccctttc
     3241 tgatttctgg catagaagaa cccccgtgaa ctgtgtggtt ccgggggttg ctgatttttg
     3301 cgagacttct cgcgcaattc cctagcttag gtgaaaacac catgaaacac tagggaaaca
     3361 cccatgaaac acccattagg gcagtagggc ggcttcttcg tctagggctt gcatttgggc
     3421 ggtgatctgg tctttagcgt gtgaaagtgt gtcgtaggtg gcgtgctcaa tgcactcgaa
     3481 cgtcacgtca tttaccgggt cacggtgggc aaagagaact agtgggttag acattgtttt
     3541 cctcgttgtc ggtggtggtg agcttttcta gccgctcggt aaacgcggcg atcatgaact
     3601 cttggaggtt ttcaccgttc tgcatgcctg cgcgcttcat gtcctcacgt agtgccaaag
     3661 gaacgcgtgc ggtgaccacg acgggcttag cctttgcctg cgcttctagt gcttcgatgg
     3721 tggcttgtgc ctgcgcttgc tgcgcctgta gtgcctgttg agcttcttgt agttgctgtt
     3781 ctagctgtgc cttggttgcc atgctttaag actctagtag ctttcctgcg atatgtcatg
     3841 cgcatgcgta gcaaacattg tcctgcaact cattcattat gtgcagtgct cctgttacta
     3901 gtcgtacata ctcatattta cctagtctgc atgcagtgca tgcacatgca gtcatgtcgt
     3961 gctaatgtgt aaaacatgta catgcagatt gctgggggtg cagggggcgg agccaccctg
     4021 tccatgcggg gtgtggggct tgccccgccg gtacagacag tgagcaccgg ggcacctagt
     4081 cgcggatacc ccccctaggt atcggacacg taaccctccc atgtcgatgc aaatctttaa
     4141 cattgagtac gggtaagctg gcacgcatag ccaagctagg cggccaccaa acaccactaa
     4201 aaattaatag ttcctagaca agacaaaccc ccgtgcgagc taccaactca aatgcacggg
     4261 ggccacataa cccgaagggg tttcaattga caaccatagc actagctaag acaacgggca
     4321 caacacccgc acaaactcgc actgcgcaac cccgcacaac atcgggtcta ggtaacactg
     4381 aaatagaagt gaacacctct aaggaaccgc aggtcaatga gggttctaag gtcactcgcg
     4441 ctagggcgtg gcgtaggcaa aacgtcatgt acaagatcac caatagtaag gctctggcgg
     4501 ggtgccatag gtggcgcagg gacgaagctg ttgcggtgtc ctggtcgtct aacggtgctt
     4561 cgcagtttga gggtctgcaa aactctcact ctcgctgggg gtcacctctg gctgaattgg
     4621 aagtcatggg cgaacgccgc attgagctgg ctattgctac taagaatcac ttggcggcgg
     4681 gtggcgcgct catgatgttt gtgggcactg ttcgacacaa ccgctcacag tcatttgcgc
     4741 aggttgaagc gggtattaag actgcgtact cttcgatggt gaaaacatct cagtggaaga
     4801 aagaacgtgc acggtacggg gtggagcaca cctatagtga ctatgaggtc acagactctt
     4861 gggcgaacgg ttggcacttg caccgcaaca tgctgttgtt cttggatcgt ccactgtctg
     4921 acgatgaact caaggcgttt gaggattcca tgttttcccg ctggtctgct ggtgtggtta
     4981 aggccggtat ggacgcgcca ctgcgtgagc acggggtcaa acttgatcag gtgtctacct
     5041 ggggtggaga cgctgcgaaa atggcaacct acctcgctaa gggcatgtct caggaactga
     5101 ctggctccgc tactaaaacc gcgtctaagg ggtcgtacac gccgtttcag atgttggata
     5161 tgttggccga tcaaagcgac gccggcgagg atatggacgc tgttttggtg gctcggtggc
     5221 gtgagtatga ggttggttct aaaaacctgc gttcgtcctg gtcacgtggg gctaagcgtg
     5281 ctttgggcat tgattacata gacgctgatg tacgtcgtga aatggaagaa gaactgtaca
     5341 agctcgccgg tctggaagca ccggaacggg tcgaatcaac ccgcgttgct gttgctttgg
     5401 tgaagcccga tgattggaaa ctgattcagt ctgatttcgc ggttaggcag tacgttctag
     5461 attgcgtgga taaggctaag gacgtggccg ctgcgcaacg tgtcgctaat gaggtgctgg
     5521 caagtctggg tgtggattcc accccgtgca tgatcgttat ggatgatgtg gacttggacg
     5581 cggttctgcc tactcatggg gacgctacta agcgtgatct gaatgcggcg gtgttcgcgg
     5641 gtaatgagca gactattctt cgcacccact aaaagcggca taaaccccgt tcgatatttt
     5701 gtgcgatgaa tttatggtca atgtcgcggg ggcaaactat gatgggtctt gttgttgaca
     5761 atggctgatt tcatcaggaa tggaactgtc atgctgttat gtgcctggct cctaatcaaa
     5821 gctggggaca atgggttgcc ccgttgatct gatctagttc ggattggcgg ggcttcactg
     5881 tatctggggg tggcatcgtg aatagattgc acaccgtagt gggcagtgtg cacaccatag
     5941 tggccatgag caccaccacc cccagggacg ccgacggcgc gaagctctgc gcctggtgcg
     6001 gctcggagat caagcaatcc ggcgtcggcc ggagccggga ctactgccgc cgctcctgcc
     6061 gccagcgggc gtacgaggcc cggcgccagc gcgaggcgat cgtgtccgcc gtggcgtcgg
     6121 cagtcgctcg ccgagatacg tcacgtgacg aaatgcagca gccttccatt ccgtcacgtg
     6181 acgaaactcg ggccgcaggt cagagcacgg ttccgcccgc tccggccctg ccggaccccc
     6241 ggcatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta cagcatccag
     6301 ggtgacggtg ccgaggatga cgatgagcgc attgttagat ttcatacacg gtgcctgact
     6361 gcgttagcaa tttaactgtg ataaactacc gcattaaagc ttatcgatga taagctgtca
     6421 aacatggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcac tgcccgcttt
     6481 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6541 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct
     6601 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc
     6661 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt
     6721 cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa
     6781 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga
     6841 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt
     6901 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac
     6961 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca
     7021 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt
     7081 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt
     7141 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt
     7201 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg
     7261 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg
     7321 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg
     7381 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt
     7441 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat
     7501 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc
     7561 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg caccattcga
     7621 tggtgtcaac gtaaatgcat gccgcttcgc cttcgcgcgc gaattgcaag ctgatccggg
     7681 cttatcgact gcacggtgca ccaatgcttc tggcgtcagg cagccatcgg aagctgtggt
     7741 atggctgtgc aggtcgtaaa tcactgcata attcgtgtcg ctcaaggcgc actcccgttc
     7801 tggataatgt tttttgcgcc gacatcataa cggttctggc aaatattctg aaatgagctg
     7861 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
     7921 caggaaacag aattaaaaga tatgaccatg attacgcc
//
