LOCUS       sequence-117172-        6403 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shuttle vector with gram-positive origin of replication isolated 
            from a native plasmid of Thermoanaerobacterium saccharolyticum 
            B6A-RI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6403)
  AUTHORS   Shaw AJ, Hogsett DA, Lynd LR
  TITLE     Natural competence in Thermoanaerobacter and Thermoanaerobacterium 
            species.
  JOURNAL   Appl Environ Microbiol. 2010 Jul;76(14):4713-9. doi: 
            10.1128/AEM.00402-10. Epub 2010 May 14.
  PUBMED    20472726
REFERENCE   2  (bases 1 to 6403)
  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"; doi: 
            "10.1128/AEM.00402-10"; journalName: "Appl Environ Microbiol"; date:
            "2010-07"; volume: "76"; issue: "14"; pages: "4713-9"
COMMENT     Sequence Label: pMU131
FEATURES             Location/Qualifiers
     source          1..6403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             829..837
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             2951..3745
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     primer_bind     complement(3786..3802)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3786..3802)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3799..3821)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3810..3826
                     /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)."
     promoter        complement(join(3834..3840,3841..3858,3859..3864))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3879..3900
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4017..4034)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4188..4776)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4268..4287)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4947..5738,5739..5807))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5570..5589
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5808..5912)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5980..5998
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6036..6058)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6158..6177
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6371..6393
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6385..6402
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6386..6402
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aattgacaaa gttttctatt tgtgttaaca ttgtttatat aatagtgaac agtgttaaga
       61 ttaaatgtga ggtgtttgta tggatattaa tgattataaa gagaagggac tttatttatt
      121 aagtagtatg gatgatttta ttaaaattaa tgatttgttt atgggtaaag ttgtttctcc
      181 tggctatgtt gcttcggttt ttggtgtttc caggtctact gttacacaat ggattcaaag
      241 acgtaaaatt agagctttta agtataaagg taaggaaggt gactatatgg ttatacctat
      301 tgctgatatt attgattaca aaagattgag taataatgat tttatttatg ataagttagt
      361 gaggtgattt attttatgtt tgacgatagc tatgttgtta atgagtgttc gtctaatgtt
      421 agtgaaaatg atagagattt ttgtagtttg gttggtcgtt ttatgattat taatggtata
      481 gataagttgg ttattaagat taatagaaaa tttaatagga aatctttaag tttagatttt
      541 agtgttgatt tattcccttc tatcaaagtt tctgaattag ttttttttga tgagtttaac
      601 aaaacgtgtg gtttttattt ttcttttaat tcttttacaa tttttaaggc ttttagagat
      661 gttcataatc ataataaaat atcattttat tttgcataat ttcgggtctg ggccgcagac
      721 caggcccagt gctaacaata ttaattttta atgttaggaa ttgtttaatt cttaattgtg
      781 tttttaaagg tagaataatt acccattcgc cctttagcca acaaaaatta aggaggtata
      841 aacatggata aaatggattt gattcttcaa gatgaaagac tgggtgagat atttaaagat
      901 atagatttaa cagataatga aaagagatat cttaaatggt tatggaaatg ggattatgaa
      961 acacgtgata cttttgtatc aatttttttg aagctaaaaa atggtggaaa atgatttttt
     1021 tcttatcttg atatattaga aaaaagcgta ctcacgaagt aagaatttgt aaaaaaagaa
     1081 ggggggattt ttttggatga gagtttgtac aagcagattt taagtaatat tattattact
     1141 cgtgattatt gtaaaaatgt tttagataat ataaagttca atgaaaaaat aattgattat
     1201 tatgttatgt tacaaaatga tgtttttatt gattttacta ataaaataaa ttcaataagg
     1261 gattgtaata aatattggta tttggatgtt tataaaaagc agaaaataaa ggattttaaa
     1321 aagactaatt tgtgtaaaga taagttctgt aataattgta agaaagttaa acaggcttca
     1381 agaatgcaaa aatatattcc tgaattacag aaatacaaag atggcttata tcattttata
     1441 tttactgttg aaaatgtgcc aggtagtgaa ttaagagata ctattgatag gttgtttaag
     1501 tcttttaagt catttacaag gtatttaagt ggtaatctta aaataaaagg tgttaatttt
     1561 gataaatggg gttataaagg ctgtgtaagg tctttagagg taacttatag tatgattgat
     1621 aatcatatta tgtatcatcc acacttgcat gttgcgatga tattagatcc tttttacgat
     1681 ggttttaatg ttgaaaggat gcatataatt aataagttta gttatagcta tggtgtttta
     1741 aaaaggttgt ttactgatga tgaattatta attcaaaaaa tttggtattt attgtttaat
     1801 aatattgagg ttaacatggc caatataaat aatttagagg atggttattc ttgtttagtt
     1861 aataagttta gtgattatga ttatgcggag ctgtttaagt atatttgtaa aaatactgat
     1921 gaacaaggtt tacttatgac ttatgatatt tttaaagatt tatattttgc attacataat
     1981 gttcatcaga tacaaggcta tggttgttta tataatataa gagatgatac tcaattagat
     2041 ttaaaggttg atgacattta taatgatttg attgatttat tacaagttac agaaaatcct
     2101 atacagtcta tggaaactgt acaggattta ttaaaggata ctgaatatac aataataagc
     2161 cgtaagcgta tatttaagta tctaacacaa ttatatcata aggattgata tttataccgt
     2221 ctgtcggact catgcggagg gggacttgag ggggtctccc ctcgcattgt acgacagacg
     2281 gtattattat tatacaaatt ttttttatgt aatttttttt gtgtaatttt tttatacaaa
     2341 taatatttca attcgagctc ggtacccggg gatcctctag agtcgacctg caggcatgca
     2401 agcttggctg caggtcgata aacccagcga accatttgag gtgataggta agattatacc
     2461 gaggtatgaa aacgagaatt ggacctttac agaattactc tatgaagcgc catatttaaa
     2521 aagctaccaa gacgaagagg atgaagagga tgaggaggca gattgccttg aatatattga
     2581 caatactgat aagataatat atcttttata tagaagatat cgccgtatgt aaggatttca
     2641 gggggcaagg cataggcagc gcgcttatca atatatctat agaatgggca aagcataaaa
     2701 acttgcatgg actaatgctt gaaacccagg acaataacct tatagcttgt aaattctatc
     2761 ataattgtgg tttcaaaatc ggctccgtcg atactatgtt atacgccaac tttgaaaaca
     2821 actttgaaaa agctgttttc tggtatttaa ggttttagaa tgcaaggaac agtgaattgg
     2881 agttcgtctt gttataatta gcttcttggg gtatctttaa atactgtaga aaagaggaag
     2941 gaaataataa atggctaaaa tgagaatatc accggaattg aaaaaactga tcgaaaaata
     3001 ccgctgcgta aaagatacgg aaggaatgtc tcctgctaag gtatataagc tggtgggaga
     3061 aaatgaaaac ctatatttaa aaatgacgga cagccggtat aaagggacca cctatgatgt
     3121 ggaacgggaa aaggacatga tgctatggct ggaaggaaag ctgcctgttc caaaggtcct
     3181 gcactttgaa cggcatgatg gctggagcaa tctgctcatg agtgaggccg atggcgtcct
     3241 ttgctcggaa gagtatgaag atgaacaaag ccctgaaaag attatcgagc tgtatgcgga
     3301 gtgcatcagg ctctttcact ccatcgacat atcggattgt ccctatacga atagcttaga
     3361 cagccgctta gccgaattgg attacttact gaataacgat ctggccgatg tggattgcga
     3421 aaactgggaa gaagacactc catttaaaga tccgcgcgag ctgtatgatt ttttaaagac
     3481 ggaaaagccc gaagaggaac ttgtcttttc ccacggcgac ctgggagaca gcaacatctt
     3541 tgtgaaagat ggcaaagtaa gtggctttat tgatcttggg agaagcggca gggcggacaa
     3601 gtggtatgac attgccttct gcgtccggtc gatcagggag gatatcgggg aagaacagta
     3661 tgtcgagcta ttttttgact tactggggat caagcctgat tgggagaaaa taaaatatta
     3721 tattttactg gatgaattgt tttagtacct agatttagat gtctaaaaag cttggcgtaa
     3781 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     3841 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     3901 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3961 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4021 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4081 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4141 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4201 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4261 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4321 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4381 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4441 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4501 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4561 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4621 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4681 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4741 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4801 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4861 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4921 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4981 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5041 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5101 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5161 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5221 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5281 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5341 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5401 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5461 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5521 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5581 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5641 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5701 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5761 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5821 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5881 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5941 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     6001 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     6061 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     6121 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     6181 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     6241 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     6301 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     6361 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtg
//
