LOCUS       40924_30575        7235 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pME8, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7235)
  AUTHORS   Audtho M, Ratlertkarn M, Wiwat C.
  TITLE     pME8, an improved shuttle vector for expression in Bacillus spp. and
            E. coli
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7235)
  AUTHORS   Audtho M, Ratlertkarn M, Wiwat C.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-FEB-2013) National Center for Genetic Engineering and 
            Biotechnology, National Science and Technology Development Agency, 
            113 Phaholyothin Road, Klong Nueng, Klong Luang, Pathum Thani 12120,
            Thailand
REFERENCE   3  (bases 1 to 7235)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7235)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (21-FEB-2013) National Center for Genetic Engineering and 
            Biotechnology, National Science and Technology Development Agency, 
            113 Phaholyothin Road, Klong Nueng, Klong Luang, Pathum Thani 12120,
            Thailand"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    7..105
                     /note="contains cloning sites SmaI, PstI, EcoRI, EcoRV,
                     HindIII, BglII, SalI, NotI, SacI, SacII, PstI, XbaI, XhoI, 
                     ApaI, and KpnI"
     promoter        complement(118..136)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(157..173)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(181..197)
                     /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(205..235)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(250..271)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(559..1147)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1321..2178)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(2179..2283)
                     /label=AmpR promoter
     rep_origin      2310..2765
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     2906..2922
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2929..2947
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_difference 3389
                     /replace="g"
                     /label=G > C to delete EcoRI site in pBC16
                     /note="G > C to delete EcoRI site in pBC16"
     CDS             3807..4808
                     /label=repB
                     /note="RepB replication protein"
     CDS             5011..6384
                     /label=TcR
                     /note="tetracycline efflux protein"
     misc_difference 6445
                     /replace="g"
                     /note="G > C to delete EcoRI site in DNA fragment from
                     pBC16"
     rep_origin      6504..6780
                     /label=ori L
                     /note="ori L"
ORIGIN
        1 cgatcccccg ggctgcagga attcgatatc aagcttagat ctaccgtcga cgcggccgcg
       61 agctcccgcg gctgcagtct agacctcgag ggggggcccg gtacccagct tttgttccct
      121 ttagtgaggg ttaatttcga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
      181 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
      241 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
      301 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
      361 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
      421 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
      481 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
      541 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
      601 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      661 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      721 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      781 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
      841 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
      901 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
      961 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     1021 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1081 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1141 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1201 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1261 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1321 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1381 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     1441 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     1501 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1561 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1621 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1681 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1741 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1801 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1861 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1921 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     1981 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2041 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2101 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2161 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2221 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     2281 gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc
     2341 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2401 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2461 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2521 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2581 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2641 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     2701 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     2761 aatttccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     2821 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     2881 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattgta atacgactca
     2941 ctatagggcg actagtgaat ggactgctta tagcgataaa gttaaatcag atttagaagt
     3001 accagcgaaa cgacacatga aaagtgttga agtgccaacg ggtgaaaagt ccatgtttgg
     3061 tttgggaaaa gaaataatga aaacagaaaa gaaaccaacc aaaaatgttg ttatatcgga
     3121 gcgtgattat aaaaacttag tgactgctgc gagagataac gataggttaa aacagcatgt
     3181 tagaaatctc atgagtactg atatggcgag agaatataaa aaattaagta aagaacatgg
     3241 gcaagttaaa gaaaaatata gtggtcttgt agagcgattt aatgaaaatg taaatgatta
     3301 taatgagttg cttgaagaaa acaagtcttt aaagtctaaa ataagcgatt taaagcgtga
     3361 tgtgagttta atctatgaaa gcactaagca attccttaag gaacgtacag acggcttaaa
     3421 agcctttaaa aacgttttta aggggtttgt agacaaggta aaggataaaa cagcacaatt
     3481 ccaagaaaaa cacgatttag aacctaaaaa gaacgaattt gaactaactc ataaccgaga
     3541 ggtaaaaaaa gaacgaagtc gagatcaggg aatgagttta taaaataaaa aaagcacctg
     3601 aaaaggtgtc tttttttgat ggttttgaac ttgttctttc ttatcttgat acatatagaa
     3661 ataacgtcat ttttatttta gttgctgaaa ggtgcgttga agtgttggta tgtatgtgtt
     3721 ttaaagtatt gaaaaccctt aaaattggtt gcacagaaaa accccatctg ttaaagttat
     3781 aagtgaccaa acaaataact aaatagatgg gggtttcttt taatattatg tgtcctaata
     3841 gtagcattta ttcagatgaa aaatcaaggg ttttagtgga caagacaaaa agtggaaaag
     3901 tgagaccatg gagagaaaag aaaatcgcta atgttgatta ctttgaactt ctgcatattc
     3961 ttgaatttaa aaaggctgaa agagtaaaag attgtgctga aatattagag tataaacaaa
     4021 atcgtgaaac aggcgaaaga aagttgtatc gagtgtggtt ttgtaaatcc aggctttgtc
     4081 caatgtgcaa ctggaggaga gcaatgaaac atggcattca gtcacaaaag gttgttgctg
     4141 aagttattaa acaaaagcca acagttcgtt ggttgtttct cacattaaca gttaaaaatg
     4201 tttatgatgg cgaagaatta aataagagtt tgtcagatat ggctcaagga tttcgccgaa
     4261 tgatgcaata taaaaaaatt aataaaaatc ttgttggttt tatgcgtgca acggaagtga
     4321 caataaataa taaagataat tcttataatc agcacatgca tgtattggta tgtgtggaac
     4381 caacttattt taagaataca gaaaactacg tgaatcaaaa acaatggatt caattttgga
     4441 aaaaggcaat gaaattagac tatgatccaa atgtaaaagt tcaaatgatt cgaccgaaaa
     4501 ataaatataa atcggatata caatcggcaa ttgacgaaac tgcaaaatat cctgtaaagg
     4561 atacggattt tatgaccgat gatgaagaaa agaatttgaa acgtttgtct gatttggagg
     4621 aaggtttaca ccgtaaaagg ttaatctcct atggtggttt gttaaaagaa atacataaaa
     4681 aattaaacct tgatgacaca gaagaaggcg atttgattca tacagatgat gacgaaaaag
     4741 ccgatgaaga tggattttct attattgcaa tgtggaattg ggaacggaaa aattatttta
     4801 ttaaagagta gttcaacaaa cgggccatat tgttgtataa gtgatgaaat actgaattta
     4861 aaacttagtt tatatgtggt aaaatgtttt aatcaagttt aggaggaatt aattatgaag
     4921 tgtaatgaat gtaacagggt tcaattaaaa gagggaagcg tatcattaac cctataaact
     4981 acgtctgccc tcattattgg agggtgacat atgaatacat cctattcaca atcgaattta
     5041 cgacacaacc aaattttaat ttggctttgc attttatctt tttttagcgt attaaatgaa
     5101 atggttttga acgtctcatt acctgatatt gcaaatgatt ttaataaacc acctgcgagt
     5161 acaaactggg tgaacacagc ctttatgtta accttttcca ttggaacagc tgtatatgga
     5221 aagctatctg atcaattagg catcaaaagg ttactcctat ttggaattat aataaattgt
     5281 ttcgggtcgg taattgggtt tgttggccat tctttctttt ccttacttat tatggctcgt
     5341 tttattcaag gggctggtgc agctgcattt ccagcactcg taatggttgt agttgcgcgc
     5401 tatattccaa aggaaaatag gggtaaagca tttggtctta ttggatcgat agtagccatg
     5461 ggagaaggag tcggtccagc gattggtgga atgatagccc attatattca ttggtcctat
     5521 cttctactca ttcctatgat aacaattatc actgttccgt ttcttatgaa attattaaag
     5581 aaagaagtaa ggataaaagg tcattttgat atcaaaggaa ttatactaat gtctgtaggc
     5641 attgtatttt ttatgttgtt tacaacatca tatagcattt cttttcttat cgttagcgtg
     5701 ctgtcattcc tgatatttgt aaaacatatc aggaaagtaa cagatccttt tgttgatccc
     5761 ggattaggga aaaatatacc ttttatgatt ggagttcttt gtgggggaat tatatttgga
     5821 acagtagcag ggtttgtctc tatggttcct tatatgatga aagatgttca ccagctaagt
     5881 actgccgaaa tcggaagtgt aattattttc cctggaacaa tgagtgtcat tattttcggc
     5941 tacattggtg ggatacttgt tgatagaaga ggtcctttat acgtgttaaa catcggagtt
     6001 acatttcttt ctgttagctt tttaactgct tcctttcttt tagaaacaac atcatggttc
     6061 atgacaatta taatcgtatt tgttttaggt gggctttcgt tcaccaaaac agttatatca
     6121 acaattgttt caagtagctt gaaacagcag gaagctggtg ctggaatgag tttgcttaac
     6181 tttaccagct ttttatcaga gggaacaggt attgcaattg taggtggttt attatccata
     6241 cccttacttg atcaaaggtt gttacctatg gaagttgatc agtcaactta tctgtatagt
     6301 aatttgttat tacttttttc aggaatcatt gtcattagtt ggctggttac cttgaatgta
     6361 tataaacatt ctcaaaggga tttctaaatc gttaagggat caactttggg agagagttca
     6421 aaattgatcc tttttttata acagcaattc aaatcttttt gttccattaa agggcgcgat
     6481 tgctgaataa aagatacgag agacctctct tgtatctttt ttattttgag tggttttgtc
     6541 cgttacacta gaaaaccgaa agacaataaa aattttattc ttgctgagtc tggctttcgg
     6601 taagctagac aaaacggaca aaataaaaat tggcaagggt ttaaaggtgg agattttttg
     6661 agtgatcttc tcaaaaaata ctacctgtcc cttgctgatt tttaaacgag cacgagagca
     6721 aaacccccct ttgctgaggt ggcagagggc aggttttttt gtttcttttt tctcgtaaaa
     6781 aaaagaaagg tcttaaaggt tttatggttt tggtcggcac tgccgacagc ctcgcagagc
     6841 acacacttta tgaatataaa gtatagtgtg ttatacttta cttggaagtg gttgccggaa
     6901 agagcgaaaa tgcctcacat ttgtgccacc taaaaaggag cgatttacat atgagttatg
     6961 cagtttgtag aatgcaaaaa gtgaaatcag ctggactaaa aggcatgcaa tttcataatc
     7021 aaagagagcg aaaaagtaga acgaatgatg atattgacca tgagcgaaca cgtgaaaatt
     7081 atgatttgaa aaatgataaa aatattgatt acaacgaacg tgtcaaagaa attattgaat
     7141 cacaaaaaac aggtacaaga aaaacgagga aagatgctgt tcttgtaaat gagttgctag
     7201 taacatctga ccgagatttt tttgagcaac tggat
//
