LOCUS       40924_43163        7520 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle expression vector pTGR5, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7520)
  AUTHORS   Ravasi P, Peiru S, Gramajo H, Menzella HG.
  TITLE     Design and testing of a synthetic biology framework for genetic 
            engineering of Corynebacterium glutamicum
  JOURNAL   Microb. Cell Fact. 11 (1), 147 (2012)
  PUBMED    23134565
REFERENCE   2  (bases 1 to 7520)
  AUTHORS   Ravasi P, Peiru S, Gramajo H, Menzella HG.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 7520)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7520)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Microb. 
            Cell Fact."; date: "2012"; volume: "11"; issue: "1"; pages: "147"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-AUG-2012) Genetic Engineering "
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Seqbuilder v. 7
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7520
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      24..31
                     /label=from Corynebacterium glutamicum sod gene
                     /note="from Corynebacterium glutamicum sod gene"
                     /regulatory_class="ribosome_binding_site"
     CDS             41..757
                     /label=EGFP
                     /note="enhanced GFP"
     CDS             793..810
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      940..1026
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1119..1146
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             complement(1352..1567)
                     /codon_start=1
                     /gene="per"
                     /product="Per"
                     /label=per
                     /note="positive effector of replication"
                     /protein_id="AFQ97508.1"
                     /translation="MDLSDVALESDALDAAVDLKTVIGFFRALDTTDAPASRDWASAAS
                     DLETLVADLEELADELRARQRQEDAQ"
     gene            complement(1352..1567)
                     /gene="per"
                     /label=per
     CDS             complement(2210..3673)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="replication protein"
                     /protein_id="AFQ97507.1"
                     /translation="MTLADPQDTVTASAWKLSADLFDTHPEAMRCGSRGWTAEDRRELL
                     AHLGRESFQGSKTRDFASAWIKNPDTGETQPKLYRAGSKALTRCQYVALTHAQHAAVIV
                     LDIDVPSHQAGGKIEHVNPQVYAILEKWARLEKAPAWIGVNPLSGKCQLIWLIDPVYAA
                     AGKTSPNMRLLAATTEEMTRVFGADQAFSHRLSRWPLHVSDDPTAYKWHCQHDRVDRLA
                     DLMEIARTMTGSQKPKKYIEQDFSSGRARIEAAQRATAEAKALAILDASLPSALDASGD
                     LIDGVRVLWTNPERARDETAFRHALTVGYQLKAAGERLKDAKIIDAYEVAYNVAQAVGA
                     DGREPDLPAMRDRLTMARRVRGYVAKGQPVVPARRVETQSSRGRKALATMGRRGAATSN
                     ARRWADPESKYAQETRQRLAEANKRREMTGELLELRVKTAILDARSQSVADPSTRELAG
                     ELGVSERRIQQVRKALGMEAKRGRPRAEN"
     gene            complement(2210..3673)
                     /gene="repA"
                     /label=repA
     CDS             4203..4994
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      5097..5685
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(5965..5986)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(6002..7081)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(7082..7159)
                     /label=lacI promoter
     promoter        7482..7510
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
ORIGIN
        1 tgagcggata acaatttgct agcgaaagga ttttttacat atggtgagca agggcgagga
       61 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
      121 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
      181 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
      241 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
      301 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
      361 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
      421 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
      481 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
      541 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
      601 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
      661 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
      721 cgccgggatc actctcggca tggacgagct gtacaagtaa agcggccgct taaggtaccg
      781 aattctaagc ttcaccacca ccaccaccac tgacctaggt gcctggcggc agtagcgcgg
      841 tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg
      901 tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag
      961 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     1021 acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggaag ggtggcgggc
     1081 aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg
     1141 cctttttact agtatatatt aattaaatat ggtaccttta attccgctag atgacgtgcg
     1201 gcttcgacct cctgggcgtg gcgcttgttg gcgcgctcgc ggctggctgc ggcacgacac
     1261 gcgtctgagc agtattttgc gcgccgtcct cgtgggtcag gccggggtgg gatcaggcca
     1321 ccgcagtagg cgcagctgat gcgatcctcc actactgcgc gtcctcctgg cgctgccgag
     1381 cacgcagctc gtcggccagc tcttcaaggt cggccacaag cgtttctagg tcgctcgcgg
     1441 cacttgccca gtcgcgtgat gctggcgcgt ctgtcgtatc gagggcgcgg aaaaatccga
     1501 tcaccgtttt taaatcgacg gcggcatcga gtgcgtcgga ctccagcgcg acatcggaga
     1561 gatccaccgc tgatgcttca ggccagtttt ggtacttcgt cgtgaaggtc atgacaccat
     1621 tataacgaac gttcgttaaa aattctagcc ccaattctga taatttcttc cggcactcct
     1681 gcgaaaacct gcgagacttc ttgcccagaa aaaacgccaa gcgcagcggt taccgcactt
     1741 tttttccagg tgatttcacc ctgaccagcg aagcggcact ttagtgcatg aggtgtgccc
     1801 ctggtttccc ctctttggag ggttcaaccc aaaaaagcac acaagcaaaa atgaaaatca
     1861 tcatgagcaa gttggtgcga agcagcaacg cgcaagctcc aaaaaggtct ccaggatctc
     1921 gaggagattt ttgaggggga gggagtcgag gaagagccag agcagaaggc ggggaaccgt
     1981 tctctgccga cagcgtgagc cccccttaaa aatcaggccg gggaggaacc ggggagggat
     2041 cagagctagg agcgagacac cctaaagggg gggaaccgtt ttctgctgac ggtgtttcgt
     2101 ttattagttt tcagcccgtg gatagcggag ggtgagggca agtgagagcc agagcaagga
     2161 cgggacccct aaagggggga accgttttct gctgacggtg tttcgtttat tagttttcag
     2221 cccgtggacg gccgcgttta gcttccattc caagtgcctt tctgacttgt tggatgcgcc
     2281 tttcactgac acctagttcg cctgcaagct cacgagtcga gggatcagca accgattgag
     2341 aacgggcatc caggatcgca gttttgacgc gaagttcgag caactcgcct gtcatttctc
     2401 ggcgtttgtt tgcttccgct aatcgctgtc gcgtctcctg cgcatactta ctttctgggt
     2461 cagcccatct gcgtgcattc gatgtagctg cgccccgtcg ccccatcgtc gctagagctt
     2521 tccgccctcg gctgctctgc gtttccaccc gacgagcagg gacgactggc tggcctttag
     2581 ccacgtagcc gcgcacacga cgcgccatcg tcaggcgatc acgcatggcg ggaagatccg
     2641 gctcccggcc gtctgcaccg accgcctggg caacgttgta cgccacttca tacgcgtcga
     2701 tgatcttggc atcttttagg cgctcaccag cagctttgag ctggtatccc acggtcaacg
     2761 cgtggcgaaa cgcggtctcg tcgcgcgctc gctctggatt tgtccagagc actcgcacgc
     2821 cgtcgatcag gtcgccggac gcgtccaggg cgctcggcag gctcgcgtcc aaaatcgcta
     2881 gtgccttggc ttctgcggtg gcgcgttgtg ccgcttcaat gcgggcgcgt ccgctggaaa
     2941 agtcctgctc aatgtacttt ttcggcttct gtgatccggt catcgttcga gcaatctcca
     3001 ttaggtcggc cagccgatcc acacgatcat gctggcagtg ccatttatag gctgtcggat
     3061 cgtctgagac gtgcagcggc caccggctca gcctatgcga aaaagcctgg tcagcgccga
     3121 aaacacgagt catttcttcc gtcgttgcag ccagcaggcg catatttggg ctggttttac
     3181 ctgctgcggc atacaccggg tcaatgagcc agatgagctg gcatttcccg ctcagcggat
     3241 tcacgccgat ccaagccggc gctttttcta ggcgtgccca tttctctaaa atcgcgtaga
     3301 cctgcgggtt tacgtgctca atcttcccgc cggcctggtg gctgggcaca tcgatgtcaa
     3361 gcacgatcac cgcggcatgt tgcgcgtgcg tcagcgcaac gtactggcac cgcgtcagcg
     3421 cttttgagcc agcccggtag agctttggtt gggtttcgcc ggtatccggg tttttaatcc
     3481 aggcgctcgc gaaatctctt gtcttgctgc cctggaagct ctcgcgtccc aggtgagcga
     3541 gcagttcgcg gcgatcttct gccgtccagc cgcgtgagcc gcagcgcata gcttcggggt
     3601 gggtgtcgaa cagatcggcg gacaatttcc acgcgcttgc tgtgactgtg tcctgcggat
     3661 cggctagagt catgtcttga gtgctttctc ccagctgatg actgggggtt agccgacgcc
     3721 ctgtgagttc ccgctcacgg ggcgttcaac tttttcaggt atttgtgcag cttatcgtgt
     3781 tttcttcgta aatgaacgct taactacctt gttaaacgtg gcaaataggc aggattgatg
     3841 gggatctagc ttcacgctgc cgcaagcact cagggcgcaa gggctgctaa aggaagcgga
     3901 acacgtagaa agccagtccg cagaaacggt gctgaccccg gatgaatgtc agctactggg
     3961 ctatctggac aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc agtgggctta
     4021 catggcgata gctagactgg gcggttttat ggacagcaag cgaaccggaa ttgccagctg
     4081 gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc
     4141 caaggatctg atggcgcagg ggatcaaggt ctgatcaaga gacaggatga ggatcgtttc
     4201 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4261 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4321 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4381 tccaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4441 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4501 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4561 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4621 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4681 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cggatgcccg
     4741 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4801 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4861 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4921 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4981 ttgacgagtt cttctgagcg ggactctggg gttcgcggaa tcatgaccaa aatcccttaa
     5041 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5101 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5161 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5221 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     5281 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5341 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5401 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5461 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5521 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5581 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5641 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5701 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5761 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     5821 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     5881 tattttctcc ttacgcatct gtgcggtatt tcacaccgca taaaagatct ctctcaaggg
     5941 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     6001 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6061 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     6121 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     6181 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     6241 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc
     6301 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     6361 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     6421 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     6481 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     6541 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     6601 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     6661 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     6721 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     6781 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     6841 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     6901 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     6961 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     7021 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     7081 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     7141 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
     7201 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatga
     7261 tatggatcca tatatgcggc cgcatattct agaatccgga gcttatcgac tgcacggtgc
     7321 accaatgctt ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa
     7381 atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc
     7441 cgacatcata acggttctgg caaatattct gaaatgagct gttgacaatt aatcatcggc
     7501 tcgtataatg tgtggaattg
//
