LOCUS       62056_8735        7054 bp DNA     circular SYN 22-JUN-2020
DEFINITION  Shuttle expression vector pEC-XK99-Gold, complete sequence.
ACCESSION   MT521917
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7054)
  AUTHORS   Walter T, Al Medani N, Burgardt A, Cankar K, Ferrer L, Kerbs A, Lee 
            JH, Mindt M, Risse JM, Wendisch VF.
  TITLE     Fermentative N-Methylanthranilate Production by Engineered 
            Corynebacterium glutamicum
  JOURNAL   Microorganisms 8 (6), E866 (2020)
  PUBMED    32521697
REFERENCE   2  (bases 1 to 7054)
  AUTHORS   Cankar K, Walter T.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-MAY-2020) Chair of Genetics of Prokaryotes, Bielefeld 
            University, Universitaetsstrasse 25, Bielefeld, NRW 33615, Germany
REFERENCE   3  (bases 1 to 7054)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Microorganisms"; date: "2020"; volume: "8"; issue: "6"; pages: 
            "E866"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (26-MAY-2020) Chair of Genetics of Prokaryotes, Bielefeld 
            University, Universitaetsstrasse 25, Bielefeld, NRW 33615, Germany"
FEATURES             Location/Qualifiers
     source          1..7054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      294..380
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      472..499
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     gene            complement(738..953)
                     /gene="per"
                     /label=per
     CDS             complement(738..953)
                     /codon_start=1
                     /transl_table=11
                     /gene="per"
                     /product="Per"
                     /label=per
                     /note="positive effector of replication"
                     /protein_id="QKU37006.1"
                     /translation="MDLSDVALESDALDAAVDLKTVIGFFRALDTTDAPASRDWASAAS
                     DLETLVADLEELADELRARQRQEDAQ"
     gene            complement(1596..3062)
                     /gene="repA"
                     /label=repA
     CDS             complement(1596..3062)
                     /codon_start=1
                     /transl_table=11
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /protein_id="QKU37007.1"
                     /translation="MTLADPQDTVTASAWKLSADLFDTHPEAMRCGSRGWTAEDRRELL
                     AHLGRESFQGSKTRDFASAWIKNPDTGETQPKLYRAGSKALTRCQYVALTHAQHAAVIV
                     LDIDVPSHQAGGKIEHVNPQVYAILEKWARLEKAPAWIGVNPLSGKCQLIWLIDPVYAA
                     AGKTSPNMRLLAATTEEMTRVFGADQAFSHRLSRWPLHVSDDPTAYKWHCQHDRVDRLA
                     DLMEIARTMTGSQKPKKYIEQDFSSGRARIEAAQRATAEAKALAILDASLPSALDASGD
                     LIDGVRVLWTNPERAARDETAFRHALTVGYQLKAAGERLKDAKIIDAYEVAYNVAQAVG
                     ADGREPDLPAMRDRLTMARRVRGYVAKGQPVVPARRVETQSSRGRKALATMGRRGAATS
                     NARRWADPESKYAQETRQRLAEANKRREMTGELLELRVKTAILDARSQSVADPSTRELA
                     GELGVSERRIQQVRKALGMEAKRGRPRAEN"
     CDS             3592..4383
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      4486..5074
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(5260..5400)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        5586..5663
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             5664..6743
                     /label=lacI
                     /note="lac repressor"
     promoter        6978..7007
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    7015..7031
                     /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 gaattccaga tgagaccgca tgcctgcaag gtctcagtat gagctcggta cccggggatc
       61 ctctagagtc gacctgcagg catgcaagct tggctgtttt ggcggatgag agaagatttt
      121 cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg
      181 cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag
      241 cgccgatggt agtgtggcgt ctccccatgc gagagtaggg aactgccagg catcaaataa
      301 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
      361 ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg
      421 gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca
      481 tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat
      541 tcaaatatgt atccgctcat gaattaattc cgctagatga cgtgcggctt cgacctcctg
      601 ggcgtggcgc ttgttggcgc gctcgcggct ggctgcggca cgacacgcgt ctgagcagta
      661 ttttgcgcgc cgtcctcgtg ggtcaggccg gggtgggatc aggccaccgc agtaggcgca
      721 gctgatgcga tcctccacta ctgcgcgtcc tcctggcgct gccgagcacg cagctcgtcg
      781 gccagctctt caaggtcggc cacaagcgtt tctaggtcgc tcgcggcact tgcccagtcg
      841 cgtgatgctg gcgcgtctgt cgtatcgagg gcgcggaaaa atccgatcac cgtttttaaa
      901 tcgacggcgg catcgagtgc gtcggactcc agcgcgacat cggagagatc caccgctgat
      961 gcttcaggcc agttttggta cttcgtcgtg aaggtcatga caccattata acgaacgttc
     1021 gttaaaaatt ctagccccaa ttctgataat ttcttccggc actcctgcga aaacctgcga
     1081 gacttcttgc ccagaaaaaa cgccaagcgc agcggttacc gcactttttt tccaggtgat
     1141 ttcaccctga ccagcgaagc ggcactttag tgcatgaggt gtgcccctgg tttcccctct
     1201 ttggagggtt caacccaaaa aagcacacaa gcaaaaatga aaatcatcat gagcaagttg
     1261 gtgcgaagca gcaacgcgct agctccaaaa acgtctccag gatctcgagg agatttttga
     1321 gggggaggga gtcgaggaag agccagagca gaaggcgggg gaaccgttct ctgccgacag
     1381 cgtgagcccc cttaaaaatc aggccgggga ggaaccgggg agggatcaga gctaggagcg
     1441 agacacccta aaggggggga accgttttct gctgacggtg tttcgtttat tagttttcag
     1501 cccgtggata gcggagggtg agggcaagtg agagccagag caaggacggg acccctaaag
     1561 gggggaaccg ttttctgctg acggtgtttc gtttattagt tttcagcccg tggacggccg
     1621 cgtttagctt ccattccaag tgcctttctg acttgttgga tgcgcctttc actgacacct
     1681 agttcgcctg caagctcacg agtcgaggga tcagcaaccg attgagaacg ggcatccagg
     1741 atcgcagttt tgacgcgaag ttcgagcaac tcgcctgtca tttctcggcg tttgtttgct
     1801 tccgctaatc gctgtcgcgt ctcctgcgca tacttacttt ctgggtcagc ccatctgcgt
     1861 gcattcgatg tagctgcgcc ccgtcgcccc atcgtcgcta gagctttccg ccctcggctg
     1921 ctctgcgttt ccacccgacg agcagggacg actggctggc ctttagccac gtagccgcgc
     1981 acacgacgcg ccatcgtcag gcgatcacgc atggcgggaa gatccggctc ccggccgtct
     2041 gcaccgaccg cctgggcaac gttgtacgcc acttcatacg cgtcgatgat cttggcatct
     2101 tttaggcgct caccagcagc tttgagctgg tatcccacgg tcaacgcgtg gcgaaacgca
     2161 gtctcgtcgc gcgctgcgcg ctctggattt gtccagagca ctcgcacgcc gtcgatcagg
     2221 tcgccggacg cgtccagggc gctcggcagg ctcgcgtcca aaatcgctag cgccttggct
     2281 tctgcggtgg cgcgttgtgc cgcttcaatg cgggcgcgtc cgctggaaaa gtcctgctca
     2341 atgtactttt tcggcttctg tgatccggtc atcgttcgag caatctccat taggtcggcc
     2401 agccgatcca cacgatcatg ctggcagtgc catttatagg ctgtcggatc gtctgagacg
     2461 tgcagcggcc accggctcag cctatgcgaa aaagcctggt cagcgccgaa aacacgagtc
     2521 atttcttccg tcgttgcagc cagcaggcgc atatttgggc tggttttacc tgctgcggca
     2581 tacaccgggt caatgagcca gatgagctgg catttcccgc tcagcggatt cacgccgatc
     2641 caagccggcg ctttttctag gcgtgcccat ttctctaaaa tcgcgtagac ctgcgggttt
     2701 acgtgctcaa tcttcccgcc ggcctggtgg ctgggcacat cgatgtcaag cacgatcacc
     2761 gcggcatgtt gcgcgtgcgt cagcgcaacg tactggcacc gcgtcagcgc ttttgagcca
     2821 gcccggtaga gctttggttg ggtttcgccg gtatccgggt ttttaatcca ggcgctcgcg
     2881 aaatctcttg tcttgctgcc ctggaagctt tcgcgtccca ggtgagcgag cagttcgcgg
     2941 cgatcttctg ccgtccagcc gcgtgagccg cagcgcatag cttcggggtg ggtgtcgaac
     3001 agatcggcgg acaatttcca cgcgctagct gtgactgtgt cctgcggatc ggctagagtc
     3061 atgtcttgag tgctttctcc cagctgatga ctgggggtta gccgacgccc tgtgagttcc
     3121 cgctcacggg gcgttcaact ttttcaggta tttgtgcagc ttatcgtgtt ttcttcgtaa
     3181 atgaacgctt aactaccttg ttaaacgtgg caaataggca ggattgatgg ggatctagct
     3241 tcacgctgcc gcaagcactc agggcgcaag ggctgctaaa ggaagcggaa cacgtagaaa
     3301 gccagtccgc agaaacggtg ctgaccccgg atgaatgtca gctactgggc tatctggaca
     3361 agggaaaacg caagcgcaaa gagaaagcag gtagcttgca gtgggcttac atggcgatag
     3421 ctagactggg cggttttatg gacagcaagc gaaccggaat tgccagctgg ggcgccctct
     3481 ggtaaggttg ggaagccctg caaagtaaac tggatggctt tcttgccgcc aaggatctga
     3541 tggcgcaggg gatcaagatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     3601 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3661 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3721 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact ccaagacgag
     3781 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3841 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3901 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3961 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4021 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4081 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc ggatgcccga cggcgaggat
     4141 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4201 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4261 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4321 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4381 ttctgagcgg gactctgggg ttcgcggaat catgaccaaa atcccttaac gtgagttttc
     4441 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4501 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4561 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4621 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4681 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4741 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4801 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4861 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4921 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4981 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5041 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5101 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5161 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     5221 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     5281 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
     5341 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     5401 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     5461 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     5521 tcaccgaaac gcgcgaggca gcagatcaat tcgcgcgcga aggcgaagcg gcatgcattt
     5581 acgttgacac catcgaatgg tgcaaaacct ttcgcggtat ggcatgatag cgcccggaag
     5641 agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg
     5701 ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga
     5761 aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg
     5821 cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc
     5881 tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca
     5941 gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca
     6001 atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg
     6061 ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc
     6121 agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc
     6181 atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct
     6241 cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga
     6301 tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc
     6361 tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg
     6421 caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat
     6481 acgacgatac cgaagacagc tcatgttata tcccgccgtc aaccaccatc aaacaggatt
     6541 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
     6601 tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca
     6661 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     6721 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gcgcgaattg
     6781 atctggtttg acagcttatc atcgactgca cggtgcacca atgcttctgg cgtcaggcag
     6841 ccatcggaag ctgtggtatg gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc
     6901 aaggcgcact cccgttctgg ataatgtttt ttgcgccgac atcataacgg ttctggcaaa
     6961 tattctgaaa tgagctgttg acaattaatc atccggctcg tataatgtgt ggaattgtga
     7021 gcggataaca atttcacaca ggaaacagac catg
//
