LOCUS       40924_14745        6984 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pDL276, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6984)
  AUTHORS   Dunny GM, Lee LN, LeBlanc DJ.
  TITLE     Improved electroporation and cloning vector system for gram-positive
            bacteria
  JOURNAL   Appl. Environ. Microbiol. 57 (4), 1194-1201 (1991)
  PUBMED    1905518
REFERENCE   2  (bases 1 to 6984)
  AUTHORS   Dunny GM, Bae T.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-DEC-1999) Microbiology, University of Minnesota, Box 
            196 UMHC, 1460 Mayo Building, 420 Delaware St. SE, Minneapolis, MN 
            55455, USA
REFERENCE   3  (bases 1 to 6984)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6984)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "1991"; volume: "57"; issue: "4"; pages:
            "1194-1201"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (15-DEC-1999) Microbiology, University of Minnesota, Box 196 UMHC, 
            1460 Mayo Building, 420 Delaware St. SE, Minneapolis, MN 55455, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6984
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(237..825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             1746..2537
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     terminator      5406..5433
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      5565..5608
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      complement(5904..5931)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(6023..6109)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     6583..6599
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    6600..6656
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(6669..6685)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6693..6709)
                     /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(6717..6747)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6762..6783)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
ORIGIN
        1 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
       61 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      121 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      181 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      241 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      301 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      361 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      421 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      481 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      541 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      601 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      661 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      721 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      781 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
      841 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
      901 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
      961 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1021 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     1081 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     1141 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     1201 agaagtggtc cgataaaccc agcgaaccat ttgaggtgat aggtaagatt ataccgaggt
     1261 atgaaaacga gaattggacc tttacagaat tactctatga agcgccatat ttaaaaagct
     1321 accaagacga agaggatgaa gaggatgagg aggcagattg ccttgaatat attgacaata
     1381 ctgataagat aatatatctt ttatatagaa gatatcgccg tatgtaagga tttcaggggg
     1441 caaggcatag gcagcgcgct tatcaatata tctatagaat gggcaaagca taaaaacttg
     1501 catggactaa tgcttgaaac ccaggacaat aaccttatag cttgtaaatt ctatcataat
     1561 tgtggtttca aaatcggctc cgtcgatact atgttatacg ccaactttca aaacaacttt
     1621 gaaaaagctg ttttctggta tttaaggttt tagaatgcaa ggaacagtga attggagttc
     1681 gtcttgttat aattagcttc ttggggtatc tttaaatact gtagaaaaga ggaaggaaat
     1741 aataaatggc taaaatgaga atatcaccgg aattgaaaaa actgatcgaa aaataccgct
     1801 gcgtaaaaga tacggaagga atgtctcctg ctaaggtata taagctggtg ggagaaaatg
     1861 aaaacctata tttaaaaatg acggacagcc ggtataaagg gaccacctat gatgtggaac
     1921 gggaaaagga catgatgcta tggctggaag gaaagctgcc tgttccaaag gtcctgcact
     1981 ttgaacggca tgatggctgg agcaatctgc tcatgagtga ggccgatggc gtcctttgct
     2041 cggaagagta tgaagatgaa caaagccctg aaaagattat cgagctgtat gcggagtgca
     2101 tcaggctctt tcactccatc gacatatcgg attgtcccta tacgaatagc ttagacagcc
     2161 gcttagccga attggattac ttactgaata acgatctggc cgatgtggat tgcgaaaact
     2221 gggaagaaga cactccattt aaagatccgc gcgagctgta tgatttttta aagacggaaa
     2281 agcccgaaga ggaacttgtc ttttcccacg gcgacctggg agacagcaac atctttgtga
     2341 aagatggcaa agtaagtggc tttattgatc ttgggagaag cggcagggcg gacaagtggt
     2401 atgacattgc cttctgcgtc cggtcgatca gggaggatat cggggaagaa cagtatgtcg
     2461 agctattttt tgacttactg gggatcaagc ctgattggga gaaaataaaa tattatattt
     2521 tactggatga attgttttag tacctagatt tagatgtcta aaaagcttta actacaagct
     2581 ttttagacat ctaatctttt ctgaagtaca tccgcaactg tccatactct gatgttttat
     2641 atcttttcta aaagttcgct agataggggt cccgagcgcc tacgaggaat ttgtatcggt
     2701 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     2761 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     2821 tttctcgtcc atatcggaaa ctactttttt attccctttt tttcgaccga gattattttg
     2881 cgataaaatc cgattaagat actgcctact aactcccaat tcttgagcta attctgatac
     2941 tgttttactc atcagctacc tcacaaaata attctttcta tcccaattcc aaaaggcaac
     3001 aatttcacga ccagtacttt ctgcagattc ttctgctccg gtctgaacaa gattcccttc
     3061 ttcgacatca tcaagagcta actccgcctt gattttcttg aacaactttc caaaaccaat
     3121 ctgccttttt cggtgcaaac cattcgttaa atcttctgtt atccgttctt tgaccatttc
     3181 agagaataat gtcacatctt ctgtatctac atcaaaaggc ttaacaggat atttagctgt
     3241 ttcgataata gctgacttca agtttttacg tttatgagct ttgacggttc gaatatcaac
     3301 tacaggtgta taatccaatt tcatcgcctt agcccatagt ttcgtccatt cttcttggct
     3361 aatgtaattt gtattatttc cagtaaaata gctagatttc acaaataaca atacatgcaa
     3421 atgagggtga taattctcat gctcagttga ataagttacc tcagtggctc gcaaatagcc
     3481 aataacattt ttatcgactt tcttgtattt cataagacga ttaaaagctc gccctatttc
     3541 tgaaattgat ttatttaatt tttcaccact aatgttctta atcgtcaagg tcaaaaagag
     3601 aaagcgacct tttggatagc gtttcattgc ttcatttacc actaattcag cttgataagc
     3661 atacttcatt gaccgtctcc agttacaaat tgggcagagc ttatttttgc aaaaataaga
     3721 ctgatagagt tttttcgtgc cgtcttgttg ttcgataaac ttcaacactt cggcacattg
     3781 atagacccgt tcaaacgaac gatatcctaa acgctctaaa cgaccagcaa gctcaatatt
     3841 ttttaacttg cgttctcgcc actttctatc tttgccattc ttagaatagt ccttaaagac
     3901 ttgattttca gtacctgtca tgttataatt ttcttgcaaa aaaatttttt tgagtatata
     3961 aaaaagttcc tgcaattctc ctaagtttat tattttaaat tggacactta aattataaac
     4021 tttattttat taaattgcaa gggctttttt gttacctaaa aacccagcaa tatcaagggt
     4081 ttgaggtgct ttaaaacaag aaaataaaaa aactcattat ttctatatgt atcaagataa
     4141 gaaaaaaccc ttgctacgca attgggtttt taagggcttt tcagccctct ttttcgtttc
     4201 acgaaaaatt cacccctaaa acctcaaatt ttgactttct aaatccaaac gtggtataat
     4261 aattttaacg ctagaagttc taggaaacga atagcacaac gcaaaaaaaa cagtagctgg
     4321 aactactgtc tttctttttt gtcctcaaaa agaactcagg ttaatttttg tcgcgtttgt
     4381 ttcgaacata ccgaatcaac caatcagcac ttacctctgc agcaacgcca accacaaaag
     4441 gtatcaaaac aagttctagg aaagtcttca cagcgctcac ctccaatcta agttcgtaac
     4501 ttagcgaaag aacgttgcga cctagctatt ataccatgtc cgcttacttt attccatggt
     4561 ctaattgaca acaagtaacc aagggcggac gtcctttgtc cgtgtcggct cgaaacgcta
     4621 aagcctttcg gctcgtcacg tcctagcgta ctttgccctt gcttattgtc aattagcttt
     4681 cacggcataa atcgctcaaa ggcctagccc tttttcaatc actcgtttaa ctacccttac
     4741 gattggctga atagctctta actctgatat aaaatttttt aaggctttat ctggcataaa
     4801 ctctttcagt tttcctagca ttttgtcatt ttgctgtctc aaaactgagt tttcacttct
     4861 aagcgctcta ttttcgttcc taagacgctc tatgctctct cccttacttt tttctttagc
     4921 tcgttccaac tcgcttcaaa ccgtttagag cgtctcagag gcactatccg ccttaattgc
     4981 tcattctcta gctctttttg atgaacaagg tttctgcttc ggtatatcag ccctttaaat
     5041 tcttcaaact gctctttagt caattccaca tattctttac cgaacacgcc ttttttaggg
     5101 gtaatagccg ctagttcttt ctcagacatg atttttaggt cagctatttg tgtttttagt
     5161 ttttctaatt tcagcccctc gcgctctaaa cgctctattt tcgcttctaa gctcttcgaa
     5221 taattttcta aggtatttat atatccctca gctttagata gctctgagag cctctcagag
     5281 gtctcagaat cgccaggacc caacgctgcc cgactgcttt cctgatgcaa aaacgaggct
     5341 agtttaccgt atctgtgggg ggatggcttg tagatatgac gacaggaaga gtttgtagaa
     5401 acgcaaaaag gccatccgtc aggatggcct tctgcttaat ttgatgcctg gcagtttatg
     5461 gcgggcgtcc tgcccgccac cctccgggcc gttgcttcgc aacgttcaaa tccgctcccg
     5521 gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc
     5581 agtctttcga ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcatgg
     5641 ggagacccca cactaccatc ggcgctacgg cgtttcactt ctgagttcgg catggggtca
     5701 ggtgggacca ccgcgctact gccgccaggc aaattctgtt ttatcagacc gcttctgcgt
     5761 tctgatttaa tctgtatcag gctgaaaatc ttctctcatc cgccaaaaca gctgctttcc
     5821 tgatgcaaaa acgaggctag tttaccgtat ctgtgggggg atggcttgta gatatgacga
     5881 caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttaattt
     5941 gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcgcaa
     6001 cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac
     6061 agataaaacg aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctggcag
     6121 ttccctactc tcgcatgggg agaccccaca ctaccatcgg cgctacggcg tttcacttct
     6181 gagttcggca tggggtcagg tgggaccacc gcgctactgc cgccaggcaa attctgtttt
     6241 atcagaccgc ttctgcgttc tgatttaatc tgtatcaggc tgaaaatctt ctctcatccg
     6301 ccaaaacagc cgagatgcgc cgcgtgcggc tgctggagat ggcggacgcg atggatatgt
     6361 tctgccaagg gttggtttgc gcattcacag ttctccgcaa gaattgattg gctccaattc
     6421 ttggagtggt gaataattcc cggcattcgc cattcaggct gcgcaactgt tgggaagggc
     6481 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     6541 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     6601 aattcgagct cggtacccgg ggatcctcta gagtcgacct gcaggcatgc aagcttggcg
     6661 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6721 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6781 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6841 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttggagcttg ttgtaactga
     6901 aaaaggaaaa ttattgtgcc aggcagttga aagtcagcac cttttaacga gtgctgaaat
     6961 gacggctaaa tgggaaacgt attt
//
