LOCUS       40924_17299        8161 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle-expression vector pEKEx2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8161)
  AUTHORS   Krumbach K, Eggeling L, Eikmanns B.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-MAR-2004) Biotechnology, Forschungszentrum, Leo-Brand,
            Juelich, NRW 52425, Germany
REFERENCE   2  (bases 1 to 8161)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8161)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (30-MAR-2004) Biotechnology, Forschungszentrum, Leo-Brand, Juelich, 
            NRW 52425, Germany"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8161
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(1..57)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(58..74)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      287..373
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      465..492
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        511..602
                     /label=AmpR promoter
     primer_bind     complement(923..939)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(947..963)
                     /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)."
     promoter        complement(971..1001)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1016..1037)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1325..1913)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             2146..2958
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             4577..5860
                     /codon_start=1
                     /product="replicase"
                     /label=replicase
                     /note="orf-1 from Corynebacterium glutamicum pBL1"
                     /protein_id="AAS99552.1"
                     /translation="MNTSKEPQVNEGSKVTRARAWRRQNVMYKITNSKALAGCHRWRRD
                     EAVAVSWSSNGASQFEGLQNSHSRWGSPLAELEVMGERRIELAIATKNHLAAGGALMMF
                     VGTVRHNRSQSFAQVEAGIKTAYSSMVKTSQWKKERARYGVEHTYSDYEVTDSWANGWH
                     LHRNMLLFLDRPLSDDELKAFEDSMFSRWSAGVVKAGMDAPLREHGVKLDQVSTWGGDA
                     AKMATYLAKGMSQELTGSATKTASKGSYTPFQMLDMLADQSDAGEDMDAVLVARWREYE
                     VGSKNLRSSWSRGAKRALGIDYIDADVRREMEEELYKLAGLEAPERVESTRVAVALVKP
                     DDWKLIQSDFAVRQYVLDCVDKAKDVAAAQRVANEVLASLGVDSTPCMIVMDDVDLDAV
                     LPTHGDATKRDLNAAVFAGNEQTILRTH"
     promoter        complement(6581..6609)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             complement(6724..7752)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ASA"
     promoter        complement(7753..7830)
                     /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        8064..8092
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    8100..8116
                     /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 gcctgcaggt cgactctaga ggatccccgg gtaccgagct cgaattcact
       61 ggccgtcgtt ttacagccaa gcttggctgt tttggcggat gagagaagat tttcagcctg
      121 atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt
      181 agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat
      241 ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa
      301 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
      361 gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg
      421 gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac
      481 ggatggcctt tttgcgtttc tacaaactct tttgtttatt tttctaaata cattcaaata
      541 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
      601 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
      661 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
      721 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
      781 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
      841 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
      901 tggttgagta attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca
      961 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     1021 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     1081 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     1141 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     1201 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     1261 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1321 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1381 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     1441 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     1501 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     1561 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     1621 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     1681 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     1741 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     1801 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     1861 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1921 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     1981 tggtcatgag attatcaaaa aggatcttca cctagatcct tttggggggg gggggaaagc
     2041 cacgttgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga
     2101 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa
     2161 cgggaaacgt cttgctcgag gccgcgatta aattccaaca tggatgctga tttatatggg
     2221 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg
     2281 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     2341 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     2401 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca
     2461 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     2521 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     2581 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     2641 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataagctt
     2701 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     2761 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     2821 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     2881 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     2941 atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt
     3001 acgctgactt gacgggacgg cggctttgtt gaataaatcg aacttttgct gagttgaagg
     3061 atcagatcac gcatcttccc gacaacgcag accgttccgt ggcaaagcaa aagttcaaaa
     3121 tcaccaactg gtccacctac aacaaagctc tcatcaaccg tggctccctc actttctggc
     3181 tggatgatgg ggcgattcag gcctggtatg agtcagcaac accttcttca cgaggcagac
     3241 ctcagcgccc ccccccccct agcttgtcta cgtctgatgc tttgaatcgg acggacttgc
     3301 cgatcttgta tgcggtgatt tttccctcgt ttgcccactt tttaatggtg gccggggtga
     3361 gagctacgcg ggcggcgacc tgctgcgctg tgatccaata ttcggggtcg ttcactggtt
     3421 cccctttctg atttctggca tagaagaacc cccgtgaact gtgtggttcc gggggttgct
     3481 gatttttgcg agacttctcg cgcaattccc tagcttaggt gaaaacacca tgaaacacta
     3541 gggaaacacc catgaaacac ccattagggc agtagggcgg cttcttcgtc tagggcttgc
     3601 atttgggcgg tgatctggtc tttagcgtgt gaaagtgtgt cgtaggtggc gtgctcaatg
     3661 cactcgaacg tcacgtcatt taccgggtca cggtgggcaa agagaactag tgggttagac
     3721 attgttttcc tcgttgtcgg tggtggtgag cttttctagc cgctcggtaa acgcggcgat
     3781 catgaactct tggaggtttt caccgttctg catgcctgcg cgcttcatgt cctcacgtag
     3841 tgccaaagga acgcgtgcgg tgaccacgac gggcttagcc tttgcctgcg cttctagtgc
     3901 ttcgatggtg gcttgtgcct gcgcttgctg cgcctgtagt gcctgttgag cttcttgtag
     3961 ttgctgttct agctgtgcct tggttgccat gctttaagac tctagtagct ttcctgcgat
     4021 atgtcatgcg catgcgtagc aaacattgtc ctgcaactca ttcattatgt gcagtgctcc
     4081 tgttactagt cgtacatact catatttacc tagtctgcat gcagtgcatg cacatgcagt
     4141 catgtcgtgc taatgtgtaa aacatgtaca tgcagattgc tgggggtgca gggggcggag
     4201 ccaccctgtc catgcggggt gtggggcttg ccccgccggt acagacagtg agcaccgggg
     4261 cacctagtcg cggatacccc ccctaggtat cggacacgta accctcccat gtcgatgcaa
     4321 atctttaaca ttgagtacgg gtaagctggc acgcatagcc aagctaggcg gccaccaaac
     4381 accactaaaa attaatagtt cctagacaag acaaaccccc gtgcgagcta ccaactcata
     4441 tgcacggggg ccacataacc cgaaggggtt tcaattgaca accatagcac tagctaagac
     4501 aacgggcaca acacccgcac aaactcgcac tgcgcaaccc cgcacaacat cgggtctagg
     4561 taacactgaa atagaagtga acacctctaa ggaaccgcag gtcaatgagg gttctaaggt
     4621 cactcgcgct agggcgtggc gtaggcaaaa cgtcatgtac aagatcacca atagtaaggc
     4681 tctggcgggg tgccataggt ggcgcaggga cgaagctgtt gcggtgtcct ggtcgtctaa
     4741 cggtgcttcg cagtttgagg gtctgcaaaa ctctcactct cgctgggggt cacctctggc
     4801 tgaattggaa gtcatgggcg aacgccgcat tgagctggct attgctacta agaatcactt
     4861 ggcggcgggt ggcgcgctca tgatgtttgt gggcactgtt cgacacaacc gctcacagtc
     4921 atttgcgcag gttgaagcgg gtattaagac tgcgtactct tcgatggtga aaacatctca
     4981 gtggaagaaa gaacgtgcac ggtacggggt ggagcacacc tatagtgact atgaggtcac
     5041 agactcttgg gcgaacggtt ggcacttgca ccgcaacatg ctgttgttct tggatcgtcc
     5101 actgtctgac gatgaactca aggcgtttga ggattccatg ttttcccgct ggtctgctgg
     5161 tgtggttaag gccggtatgg acgcgccact gcgtgagcac ggggtcaaac ttgatcaggt
     5221 gtctacctgg ggtggagacg ctgcgaaaat ggcaacctac ctcgctaagg gcatgtctca
     5281 ggaactgact ggctccgcta ctaaaaccgc gtctaagggg tcgtacacgc cgtttcagat
     5341 gttggatatg ttggccgatc aaagcgacgc cggcgaggat atggacgctg ttttggtggc
     5401 tcggtggcgt gagtatgagg ttggttctaa aaacctgcgt tcgtcctggt cacgtggggc
     5461 taagcgtgct ttgggcattg attacataga cgctgatgta cgtcgtgaaa tggaagaaga
     5521 actgtacaag ctcgccggtc tggaagcacc ggaacgggtc gaatcaaccc gcgttgctgt
     5581 tgctttggtg aagcccgatg attggaaact gattcagtct gatttcgcgg ttaggcagta
     5641 cgttctagat tgcgtggata aggctaagga cgtggccgct gcgcaacgtg tcgctaatga
     5701 ggtgctggca agtctgggtg tggattccac cccgtgcatg atcgttatgg atgatgtgga
     5761 cttggacgcg gttctgccta ctcatgggga cgctactaag cgtgatctga atgcggcggt
     5821 gttcgcgggt aatgagcaga ctattcttcg cacccactaa aagcggcata aaccccgttc
     5881 gatattttgt gcgatgaatt tatggtcaat gtcgcggggg caaactatga tgggtcttgt
     5941 tgttgacaat ggctgatttc atcaggaatg gaactgtcat gctgttatgt gcctggctcc
     6001 taatcaaagc tggggacaat gggttgcccc gttgatctga tctagttcgg attggcgggg
     6061 cttcactgta tctgggggtg gcatcgtgaa tagattgcac accgtagtgg gcagtgtgca
     6121 caccatagtg gccatgagca ccaccacccc cagggacgcc gacggcgcga agctctgcgc
     6181 ctggtgcggc tcggagatca agcaatccgg cgtcggccgg agccgggact actgccgccg
     6241 ctcctgccgc cagcgggcgt acgaggcccg gcgccagcgc gaggcgatcg tgtccgccgt
     6301 ggcgtcggca gtcgctcgcc gagatacgtc acgtgacgaa atgcagcagc cttccattcc
     6361 gtcacgtgac gaaactcggg ccgcaggtca gagcacggtt ccgcccgctc cggccctgcc
     6421 ggacccccgg catcccgcaa gaggcccggc agtaccggca taaccaagcc tatgcctaca
     6481 gcatccaggg tgacggtgcc gaggatgacg atgagcgcat tgttagattt catacacggt
     6541 gcctgactgc gttagcaatt taactgtgat aaactaccgc attaaagctt atcgatgata
     6601 agctgtcaaa catggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc
     6661 cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat
     6721 ggcggccgac gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt
     6781 caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag
     6841 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg
     6901 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc
     6961 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc
     7021 aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc
     7081 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag
     7141 atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa
     7201 cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc
     7261 ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc
     7321 cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt
     7381 aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc
     7441 gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga
     7501 tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc
     7561 aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag
     7621 ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt
     7681 caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt
     7741 tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc
     7801 gcgaaaggtt ttgcaccatt cgatggtgtc aacgtaaatg catgccgctt cgccttcgcg
     7861 cgcgaattgc aagctgatcc gggcttatcg actgcacggt gcaccaatgc ttctggcgtc
     7921 aggcagccat cggaagctgt ggtatggctg tgcaggtcgt aaatcactgc ataattcgtg
     7981 tcgctcaagg cgcactcccg ttctggataa tgttttttgc gccgacatca taacggttct
     8041 ggcaaatatt ctgaaatgag ctgttgacaa ttaatcatcg gctcgtataa tgtgtggaat
     8101 tgtgagcgga taacaatttc acacaggaaa cagaattaaa agatatgacc atgattacgc
     8161 c
//
