LOCUS       40924_32708        5500 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMU749, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5500)
  AUTHORS   Olson DG.
  TITLE     Transformation of Clostridium thermocellum by electroporation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5500)
  AUTHORS   Olson DG.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-OCT-2011) Thayer School of Engineering, Dartmouth 
            College, 8000 Cummings, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 5500)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5500)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-OCT-2011) Thayer School of Engineering, Dartmouth College, 8000 
            Cummings, Hanover, NH 03755, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5500
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      826..1280
                     /label=cth ori
                     /note="cth ori"
     CDS             1281..2282
                     /codon_start=1
                     /label=repB
                     /note="RepB replication protein"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     promoter        2400..2504
                     /label=AmpR promoter
     CDS             2505..3362
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     protein_bind    3524..3540
                     /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)."
     primer_bind     3548..3564
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(3631..4134)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        4162..4382
                     /label=URA3 promoter
     CDS             4383..5183
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagtatgg gaaacaaaat
      841 attgcgtatg cgactgttca catggacgag aaaacccctc acatgcattt aggagttgtt
      901 cctatgcgcc tagagggctt ttcgtgcgtc agcatgagcg atcggaagaa aagaagaata
      961 ctttttcgct tcaagatgtt ttgcagcgtg atcgagaact tcgtgagcaa agaaaagcaa
     1021 agaggaaaaa atcgcatgat ttggagcgat aagaaaaagc actcgaatga gtgctttttt
     1081 tgcgttttga gcgtagcgaa aaacgagttc tttctattct tgatacatat agaaataacg
     1141 tcatttttat tttagttgct gaaaggtgcg ttgaagtgtt ggtatgtatg tgttttaaag
     1201 tattgaaaac ccttaaaatt ggttgcacag aaaaacccca tctgttaaag ttataagtga
     1261 ccaaacaaat aactaaatag atgggggttt cttttaatat tatgtgtcct aatagtagca
     1321 tttattcaga tgaaaaatca agggttttag tggacaagac aaaaagtgga aaagtgagac
     1381 catggagaga aaagaaaatc gctaatgttg attactttga acttctgcat attcttgaat
     1441 ttaaaaaggc tgaaagagta aaagattgtg ctgaaatatt agagtataaa caaaatcgtg
     1501 aaacaggcga aagaaagttg tatcgagtgt ggttttgtaa atccaggctt tgtccaatgt
     1561 gcaactggag gagagcaatg aaacatggca ttcagtcaca aaaggttgtt gctgaagtta
     1621 ttaaacaaaa gccaacagtt cgttggttgt ttctcacatt aacagttaaa aatgtttatg
     1681 atggcgaaga attaaataag agtttgtcag atatggctca aggatttcgc cgaatgatgc
     1741 aatataaaaa aattaataaa aatcttgttg gttttatgcg tgcaacggaa gtgacaataa
     1801 ataataaaga taattcttat aatcagcaca tgcatgtatt ggtatgtgtg gaaccaactt
     1861 attttaagaa tacagaaaac tacgtgaatc aaaaacaatg gattcaattt tggaaaaagg
     1921 caatgaaatt agactatgat ccaaatgtaa aagttcaaat gattcgaccg aaaaataaat
     1981 ataaatcgga tatacaatcg gcaattgacg aaactgcaaa atatcctgta aaggatacgg
     2041 attttatgac cgatgatgaa gaaaagaatt tgaaacgttt gtctgatttg gaggaaggtt
     2101 tacaccgtaa aaggttaatc tcctatggtg gtttgttaaa agaaatacat aaaaaattaa
     2161 accttgatga cacagaagaa ggcgatttga ttcatacaga tgatgacgaa aaagccgatg
     2221 aagatggatt ttctattatt gcaatgtgga attgggaacg gaaaaattat tttattaaag
     2281 agtagttcaa caaacgggat tgacttttaa aaaaggattg attctaatga agaaagcaga
     2341 caagtaagcc tccgaattcg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     2401 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2461 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     2521 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2581 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2641 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2701 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     2761 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2821 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2881 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2941 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3001 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     3061 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     3121 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3181 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3241 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3301 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3361 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3421 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3481 gtgagttttc gttccactga gcgtcaagat ctgttgtgtg gaattgtgag cggataacaa
     3541 tttcacacag gaaacagcta tgaccatgat tacgccaagc ttgcatgccc tgcaggtcga
     3601 ctctagagga tccccgggta ccgcggccgc gatcgcttgc ctgtaactta cacgcgcctc
     3661 gtatctttta atgatggaat aatttgggaa tttactctgt gtttatttat ttttatgttt
     3721 tgtatttgga ttttagaaag taaataaaga aggtagaaga gttacggaat gaagaaaaaa
     3781 aaataaacaa aggtttaaaa aatttcaaca aaaagcgtac tttacatata tatttattag
     3841 acaagaaaag cagattaaat agatatacat tcgattaacg ataagtaaaa tgtaaaatca
     3901 caggattttc gtgtgtggtc ttctacacag acaagatgaa acaattcggc attaatacct
     3961 gagagcagga agagcaagat aaaaggtagt atttgttggc gatcccccta gagtctttta
     4021 catcttcgga aaacaaaaac tattttttct ttaatttctt tttttacttt ctatttttaa
     4081 tttatatatt tatattaaaa aatttaaatt ataattattt ttatagcacg tgatgaaaag
     4141 gacccatcga taagctagct tttcaattca attcatcatt ttttttttat tctttttttt
     4201 gatttcggtt tctttgaaat ttttttgatt cggtaatctc cgaacagaag gaagaacgaa
     4261 ggaaggagca cagacttaga ttggtatata tacgcatatg tagtgttgaa gaaacatgaa
     4321 attgcccagt attcttaacc caactgcaca gaacaaaaac ctgcaggaaa cgaagataaa
     4381 tcatgtcgaa agctacatat aaggaacgtg ctgctactca tcctagtcct gttgctgcca
     4441 agctatttaa tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta
     4501 ccaccaagga attactggag ttagttgaag cattaggtcc caaaatttgt ttactaaaaa
     4561 cacatgtgga tatcttgact gatttttcca tggagggcac agttaagccg ctaaaggcat
     4621 tatccgccaa gtacaatttt ttactcttcg aagacagaaa atttgctgac attggtaata
     4681 cagtcaaatt gcagtactct gcgggtgtat acagaatagc agaatgggca gacattacga
     4741 atgcacacgg tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg gcagaagaag
     4801 taacaaagga acctagaggc cttttgatgt tagcagaatt gtcatgcaag ggctccctat
     4861 ctactggaga atatactaag ggtactgttg acattgcgaa gagcgacaaa gattttgtta
     4921 tcggctttat tgctcaaaga gacatgggtg gaagagatga aggttacgat tggttgatta
     4981 tgacacccgg tgtgggttta gatgacaagg gagacgcatt gggtcaacag tatagaaccg
     5041 tggatgatgt ggtctctaca ggatctgaca ttattattgt tggaagagga ctatttgcaa
     5101 agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg gaagcatatt
     5161 tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca tgtatactaa
     5221 actcacaaat tagagcttca atttaattat atcagttatt acccactttt cggcggccgc
     5281 gaattgacag atcgctgaga taggtgcctc actggattaa gcattggtaa ctgtcagacc
     5341 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5401 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5461 actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
