LOCUS       sequence-100798-        3568 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning and expression vector for Clostridium thermocellum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3568)
  AUTHORS   Olson DG, Lynd LR
  TITLE     Computational design and characterization of a temperature-sensitive
            plasmid replicon for gram positive thermophiles.
  JOURNAL   J Biol Eng. 2012 May 11;6(1):5. doi: 10.1186/1754-1611-6-5.
  PUBMED    22578246
REFERENCE   2  (bases 1 to 3568)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Eng."; date: "2012-05-11"; volume: "6(1):5. doi"; pages: " 
            10.1186/1754-1611-6-5"
COMMENT     Sequence Label: pMU102
FEATURES             Location/Qualifiers
     source          1..3568
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             1281..2285
                     /codon_start=1
                     /gene="repB"
                     /product="RepB replication protein"
                     /label=repB
                     /note="from Enterococcus faecalis plasmid pAM-alpha-1"
                     /db_xref="GI:22652809"
                     /protein_id="AAN03827.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     protein_bind    3172..3193
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3208..3213,3214..3231,3232..3238)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3246..3262
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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     3251..3273
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3270..3286
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3270..3286
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
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 tcctaaattc actttagata aaaatttagg aggcatatca aatgaacttt
     2401 aataaaattg atttagacaa ttggaagaga aaagagatat ttaatcatta tttgaaccaa
     2461 caaacgactt ttagtataac cacagaaatt gatattagtg ttttataccg aaacataaaa
     2521 caagaaggat ataaatttta ccctgcattt attttcttag tgacaagggt gataaactca
     2581 aatacagctt ttagaactgg ttacaatagc gacggagagt taggttattg ggataagtta
     2641 gagccacttt atacaatttt tgatggtgta tctaaaacat tctctggtat ttggactcct
     2701 gtaaagaatg acttcaaaga gttttatgat ttataccttt ctgatgtaga gaaatataat
     2761 ggttcgggga aattgtttcc caaaacacct atacctgaaa atgctttttc tctttctatt
     2821 attccatgga cttcatttac tgggtttaac ttaaatatca ataataatag taattacctt
     2881 ctacccatta ttacagcagg aaaattcatt aataaaggta attcaatata tttaccgcta
     2941 tctttacagg tacatcattc tgtttgtgat ggttatcatg caggattgtt tatgaactct
     3001 attcaggaat tgtcagatag gcctaatgac tggcttttat aatatgagat aatgccgact
     3061 gtacttttta cagtcggttt tctaatgtca ctagggctcg cctttgggaa gtttgaaggg
     3121 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     3181 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     3241 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     3301 gcttgcatgc ctgcaggtcg actctagagg atccccgggt accgagctcg aattgacaga
     3361 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3421 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3481 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3541 accccgtaga aaagatcaaa ggatcttc
//
