LOCUS       40924_16695        7509 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle expression vector pEC-XT99A, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7509)
  AUTHORS   Kirchner O, Tauch A.
  TITLE     Tools for genetic engineering in the amino acid-producing bacterium 
            Corynebacterium glutamicum
  JOURNAL   J. Biotechnol. 104 (1-3), 287-299 (2003)
  PUBMED    12948646
REFERENCE   2  (bases 1 to 7509)
  AUTHORS   Kirchner O, Tauch A.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-JAN-2003) Department of Genetics, University of 
            Bielefeld, Universitaetsstrasse 25, Bielefeld D-33615, Germany
REFERENCE   3  (bases 1 to 7509)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7509)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Biotechnol. 104 (1-3), 287-299 (2003)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (15-JAN-2003) Department of Genetics, University of Bielefeld, 
            Universitaetsstrasse 25, Bielefeld D-33615, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7509
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..57
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      260..346
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      438..465
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             769..2232
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="replication protein"
                     /protein_id="AAO65199.1"
                     /translation="MTLADPQDTVTASAWKLSADLFDTHPEAMRCGSRGWTAEDRRELL
                     AHLGRESFQGSKTRDFASAWIKNPDTGETQPKLYRAGSKALTRCQYVALTHAQHAAVIV
                     LDIDVPSHQAGGKIEHVNPQVYAILEKWARLEKAPAWIGVNPLSGKCQLIWLIDPVYAA
                     AGKTSPNMRLLAATTEEMTRVFGADQAFSHRLSRWPLHVSDDPTAYKWHCQHDRVDRLA
                     DLMEIARTMTGSQKPKKYIEQDFSSGRARIEAAQRATAEAKALAILDASLPSALDASGD
                     LIDGVRVLWTNPERARDETAFRHALTVGYQLKAAGERLKDAKIIDAYEVAYNVAQAVGA
                     DGREPDLPAMRDRLTMARRVRGYVAKGQPVVPARRVETQSSRGRKALATMGRRGAATSN
                     ARRWADPESKYAQETRQRLAEANKRREMTGELLELRVKTAILDARSQSVADPSTRELAG
                     ELGVSERRIQQVRKALGMEAKRGRPRAEN"
     gene            769..2232
                     /gene="repA"
                     /label=repA
     CDS             2875..3090
                     /codon_start=1
                     /gene="per"
                     /product="Per"
                     /label=per
                     /note="positive effector of replication"
                     /protein_id="AAO65200.1"
                     /translation="MDLSDVALESDALDAAVDLKTVIGFFRALDTTDAPASRDWASAAS
                     DLETLVADLEELADELRARQRQEDAQ"
     gene            2875..3090
                     /gene="per"
                     /label=per
     CDS             complement(3651..3662)
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     rep_origin      4941..5529
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(5715..5855)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        6041..6118
                     /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             6119..7198
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        7433..7462
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    7470..7486
                     /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 gaattcgagc tcggtacccg gggatcctct agagtcgacc tgcaggcatg caagcttggc
       61 tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc
      121 ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg
      181 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga
      241 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
      301 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga
      361 tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc
      421 caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac
      481 tctttttgtt tatttttcta aatacattca aatatgtatc cgctcatgct ccttcgtcgg
      541 tgtcgtcgcc ggatggtctg cggtggtgct cagcgtggag acgcgcaccg tcacggaccc
      601 ccatcaatcc tgcctatttg ccacgtttaa caaggtagtt aagcgttcat ttacgaagaa
      661 aacacgataa gctgcacaaa tacctgaaaa agttgaacgc cccgtgagcg ggaactcaca
      721 gggcgtcggc taacccccag tcatcagctg ggagaaagca ctcaagacat gactctagcc
      781 gatccgcagg acacagtcac agctagcgcg tggaaattgt ccgccgatct gttcgacacc
      841 caccccgaag ctatgcgctg cggctcacgc ggctggacgg cagaagatcg ccgcgaactg
      901 ctcgctcacc tgggacgcga aagcttccag ggcagcaaga caagagattt cgcgagcgcc
      961 tggattaaaa acccggatac cggcgaaacc caaccaaagc tctaccgggc tggctcaaaa
     1021 gcgctgacgc ggtgccagta cgttgcgctg acgcacgcgc aacatgccgc ggtgatcgtg
     1081 cttgacatcg atgtgcccag ccaccaggcc ggcgggaaga ttgagcacgt aaacccgcag
     1141 gtctacgcga ttttagagaa atgggcacgc ctagaaaaag cgccggcttg gatcggcgtg
     1201 aatccgctga gcgggaaatg ccagctcatc tggctcattg acccggtgta tgccgcagca
     1261 ggtaaaacca gcccaaatat gcgcctgctg gctgcaacga cggaagaaat gactcgtgtt
     1321 ttcggcgctg accaggcttt ttcgcatagg ctgagccggt ggccgctgca cgtctcagac
     1381 gatccgacag cctataaatg gcactgccag catgatcgtg tggatcggct ggccgaccta
     1441 atggagattg ctcgaacgat gaccggatca cagaagccga aaaagtacat tgagcaggac
     1501 ttttccagcg gacgcgcccg cattgaagcg gcacaacgcg ccaccgcaga agccaaggcg
     1561 ctagcgattt tggacgcgag cctgccgagc gccctggacg cgtccggcga cctgatcgac
     1621 ggcgtgcgag tgctctggac aaatccagag cgagcgcgcg acgagaccgc gtttcgccac
     1681 gcgttgaccg tgggatacca gctcaaagct gctggtgagc gcctaaaaga tgccaagatc
     1741 atcgacgcgt atgaagtggc gtacaacgtt gcccaggcgg tcggtgcaga cggccgggag
     1801 ccggatcttc ccgccatgcg tgatcgcctg acgatggcgc gtcgtgtgcg cggctacgtg
     1861 gctaaaggcc agccagtcgt ccctgctcgt cgggtggaaa cgcagagcag ccgagggcgg
     1921 aaagctctag cgacgatggg gcgacggggc gcagctacat cgaatgcacg cagatgggct
     1981 gacccagaaa gtaagtatgc gcaggagacg cgacagcgat tagcggaagc aaacaaacgc
     2041 cgagaaatga caggcgagtt gctcgaactt cgcgtcaaaa ctgcgatcct ggatgcccgt
     2101 tctcaatcgg ttgctgatcc ctcgactcgt gagcttgcag gcgaactagg tgtcagtgaa
     2161 aggcgcatcc aacaagtcag aaaggcactt ggaatggaag ctaaacgcgg ccgtccacgg
     2221 gctgaaaact aataaacgaa acaccgtcag cagaaaacgg ttcccccctt taggggtccc
     2281 gtccttgctc tggctctcac ttgccctcac cctccgctat ccacgggctg aaaactaata
     2341 aacgaaacac cgtcagcaga aaacggttcc ccccctttag ggtgtctcgc tcctagctct
     2401 gatccctccc cggttcctcc ccggcctgat ttttaagggg ggctcacgct gtcggcagag
     2461 aacggttccc cgccttctgc tctggctctt cctcgactcc ctccccctca aaaatctcct
     2521 cgagatcctg gagacctttt tggagctagc gcgttgctgc ttcgcaccaa cttgctcatg
     2581 atgattttca tttttgcttg tgtgcttttt tgggttgaac cctccaaaga ggggaaacca
     2641 ggggcacacc tcatgcacta aagtgccgct tcgctggtca gggtgaaatc acctggaaaa
     2701 aaagtgcggt aaccgctgcg cttggcgttt tttctgggca agaagtctcg caggttttcg
     2761 caggagtgcc ggaagaaatt atcagaattg gggctagaat ttttaacgaa cgttcgttat
     2821 aatggtgtca tgaccttcac gacgaagtac caaaactggc ctgaagcatc agcggtggat
     2881 ctctccgatg tcgcgctgga gtccgacgca ctcgatgccg ccgtcgattt aaaaacggtg
     2941 atcggatttt tccgcgccct cgatacgaca gacgcgccag catcacgcga ctgggcaagt
     3001 gccgcgagcg acctagaaac gcttgtggcc gaccttgaag agctggccga cgagctgcgt
     3061 gctcggcagc gccaggagga cgcgcagtag tggaggatcg catcagctgc gcctactgcg
     3121 gtggcctgat cccaccccgg cctgacccac gaggacggcg cgcaaaatac tgctcagacg
     3181 cgtgtcgtgc cgcagccagc cgcgagcgcg ccaacaagcg ccacgcccag gaggtcgaag
     3241 ccgcacgtcg accgcgtgta gtgcgtggcg gaaacttctt gcgtttcgca agagaaatgc
     3301 gtcccatttc tcgtcggact cggggaagga agcgtgatgc tctcggtcaa gcacgtcgct
     3361 cgccagcgct gcgaggagtt cggccttcgt gcggaagtgc cagtagaggc cgggctgctg
     3421 tacctgtaag tgagccgcca gcgcgcgagt ggtgaagcca tcgagcccag tctcgtcgag
     3481 cacctgccgg gccccgagca acacggacgt gcggtcgaga cgcttccggt ggtgagtcat
     3541 agttgcactt tatcatcgat aactttatct tagataaagt gactgctcgc tactctcatc
     3601 tgactgctcg ctactctcat cgtggaatcc tgacagccgt gctcatcacg gcgaccctcg
     3661 atgctgcagg gctgggcctc gtgatgccga tcttgcctac ccttctcgac caggtcggtg
     3721 cccccgacga catgatccca ctgcacgtcg gactactgac agcgctctat gcgatcatgc
     3781 agtttctttg cgccccgatc cttggccgac tctctgaccg tttcggacgc cgccgcgtgc
     3841 ttgtcgcctc cctcgcaggc gcgacgatcg actacctcgt gctcgcactg acggacacgc
     3901 tgtgggtctt ttacctcgcc cgcgcggttg caggcattac cggcgccacg aacgccgtca
     3961 ccgcgacggt gatcgccgac attactccgc cggatcagcg cgcaaaacgc tacgggtggc
     4021 tcggcgcatg ctacggcggt ggcatgatcg cgggtcccgc cattggcggt cttttcggcg
     4081 gggtctcacc gcatctgcca ttcctcgtcg ccgccgcgct cgccggaatc accctcgtac
     4141 tcagcgcgag tcttctgcgt gagacgcggc caccgggcag caacggctcg cacgcacagc
     4201 aacccggtac ggcgaagcga accgcagtgc cggggatgct tatccttctc gcagtcttcg
     4261 gcatcgtgca gttcatcggc caagcaccag gctccacctg ggtgctcttc acgcagcagc
     4321 gcctcgactg gaaccccgtc gaagtcggcg tttcgctatc catcttcgga atggtgcaag
     4381 tattcgtgca ggcggcactg accggacgca tcgtgtcccg gatcggcgag acccgggcga
     4441 tcctcgtcgg tatcgccgca gacgccattg ggctcatcgg ccttgccctc atcgccagca
     4501 catgggcgat gctaccgatc ctcgcagcgc tcggactcgg cagcatcacg ttgcccgcac
     4561 tgcagacgct gctctcgaga cgcgcgcccg agcagcagca gggacgcctg cagggaacac
     4621 ttgcaagcct gaacagcctc acctcgatca tcggcccggt caccttcacc ggcattttcg
     4681 cactcacccg aacgaatgca gacggcaccc tctggatctg cgccgcagcg ctctacgttc
     4741 tctgcgccct cctgatgatc cgtgagacat gcgcctcacg gcgatctcga taaccgcgct
     4801 aaggtgccat cccgatgcga cgggatcgct ctgccaccag tcaagtctcc cgtagccggt
     4861 atgagcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4921 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4981 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5041 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     5101 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5161 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5221 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5281 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5341 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5401 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5461 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5521 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5581 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     5641 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     5701 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     5761 cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     5821 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     5881 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     5941 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagcaga
     6001 tcaattcgcg cgcgaaggcg aagcggcatg catttacgtt gacaccatcg aatggtgcaa
     6061 aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt
     6121 gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc
     6181 ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc
     6241 gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc
     6301 gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc
     6361 ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg
     6421 aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg
     6481 gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac
     6541 taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt
     6601 ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca
     6661 aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg
     6721 gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag
     6781 tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc
     6841 gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg
     6901 gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg
     6961 ttatatcccg ccgtcaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt
     7021 ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt
     7081 ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc
     7141 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     7201 agcgcaacgc aattaatgtg agttagcgcg aattgatctg gtttgacagc ttatcatcga
     7261 ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg gtatggctgt
     7321 gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt tctggataat
     7381 gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc tgttgacaat
     7441 taatcatccg gctcgtataa tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     7501 cagaccatg
//
