LOCUS       sequence-89822-e        3849 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shuttle vector: pUC ori, pTHT15 ori for use in E. coli/B. subtilis. 
            Cannot modify orientation of cat gene; promoter too weak in E. coli,
            probably transcribed via repB promoter in bicistronic manner..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3849)
  AUTHORS   Wright O, Delmans M, Stan GB, Ellis T
  TITLE     GeneGuard: a Modular Plasmid System Designed for Biosafety.
  JOURNAL   ACS Synth Biol. 2014 May 13.
  PUBMED    24847673
REFERENCE   2  (bases 1 to 3849)
  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: "ACS Synth 
            Biol. 2014 May 13."
COMMENT     Sequence Label: pSEVA3b61
FEATURES             Location/Qualifiers
     source          1..3849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..31
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    56..112
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      155..249
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     oriT            1259..1367
                     /note="incP origin of transfer"
     CDS             complement(1419..2423)
                     /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"
     primer_bind     complement(2944..2961)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3115..3703)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3195..3214)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3757..3843)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaattaaag cggataacaa tttcacacag gaggccgcct aggccgcggc cgcgcgaatt
       61 cgagctcggt acccggggat cctctagagt cgacctgcag gcatgcaagc ttgcggccgc
      121 gtcgtgactg ggaaaaccct ggcgactagt cttggactcc tgttgataga tccagtaatg
      181 acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt
      241 ggtgagaatc caggggtccc caataattac gatttaaatc cttcaaactt cccaaaggcg
      301 agccctagtg acattagaaa accgactgta aaaagtacag tcggcattat ctcatattat
      361 aaaagccagt cattaggcct atctgacaat tcctgaatag agttcataaa caatcctgca
      421 tgataaccat cacaaacaga atgatgtacc tgtaaagata gcggtaaata tattgaatta
      481 cctttattaa tgaattttcc tgctgtaata atgggtagaa ggtaattact attattattg
      541 atatttaagt taaacccagt aaatgaagtc catggaataa tagaaagaga aaaagcattt
      601 tcaggtatag gtgttttggg aaacaatttc cccgaaccat tatatttctc tacatcagaa
      661 aggtataaat cataaaactc tttgaagtca ttctttacag gagtccaaat accagagaat
      721 gttttagata caccatcaaa aattgtataa agtggctcta acttatccca ataacctaac
      781 tctccgtcgc tattgtaacc agttctaaaa gctgtatttg agtttatcac ccttgtcact
      841 aagaaaataa atgcagggta aaatttatat ccttcttgtt ttatgtttcg gtataaaaca
      901 ctaatatcaa tttctgtggt tatactaaaa gtcgtttgtt ggttcaaata atgattaaat
      961 atctcttttc tcttccaatt gtctaaatca attttattaa agttcatttg atatgcctcc
     1021 taaattttta tctaaagtga atttaggagg cttacttgtc tgctttcttc attagaatca
     1081 atcctttttt aaaagtcaat cccgtttgtt gaagactttt gtccttttcc gctgcataac
     1141 cctgcttcgg ggtcattata gcgatttttt cggtatatcc atcctttttc gcacgatata
     1201 caggattttg ccaaagggtt cgtgtagact ttccttggtg tatccaacgg cgtcagccgg
     1261 gcaggatagg tgaagtaggc ccacccgcga gcgggtgttc cttcttcact gtcccttatt
     1321 cgcacctggc ggtgctcaac gggaatcctg ctctgcgagg ctggccgtag gccggcctag
     1381 aatcaatcct tttttaaaag tcaatcccgt ttgttgaact actctttaat aaaataattt
     1441 ttccgttccc aattccacat tgcaataata gaaaatccat cttcatcggc tttttcgtca
     1501 tcatctgtat gaatcaaatc gccttcttct gtgtcatcaa ggtttaattt tttatgtatt
     1561 tcttttaaca aaccaccata ggagattaac cttttacggt gtaaaccttc ctccaaatca
     1621 gacaaacgtt tcaaattctt ttcttcatca tcggtcataa aatccgtatc ctttacagga
     1681 tattttgcag tttcgtcaat tgccgattgt atatccgatt tatatttatt tttcggtcga
     1741 atcatttgaa cttttacatt tggatcatag tctaatttca ttgccttttt ccaaaattga
     1801 atccattgtt tttgattcac gtagttttct gtattcttaa aataagttgg ttccacacat
     1861 accaatacat gcatgtgctg attataagaa ttatctttat tatttattgt cacttccgtt
     1921 gcacgcataa aaccaacaag atttttatta atttttttat attgcatcat tcggcgaaat
     1981 ccttgagcca tatctgacaa actcttattt aattcttcgc catcataaac atttttaact
     2041 gttaatgtga gaaacaacca acgaactgtt ggcttttgtt taataacttc agcaacaacc
     2101 ttttgtgact gaatgccatg tttcattgct ctcctccagt tgcacattgg acaaagcctg
     2161 gatttacaaa accacactcg atacaacttt ctttcgcctg tttcacgatt ttgtttatac
     2221 tctaatattt cagcacaatc ttttactctt tcagcctttt taaattcaag aatatgcaga
     2281 agttcaaagt aatcaacatt agcgattttc ttttctctcc atggtctcac ttttccactt
     2341 tttgtcttgt ccactaaaac ccttgatttt tcatctgaat aaatgctact attaggacac
     2401 ataatattaa aagaaacccc catctattta gttatttgtt tggtcactta taactttaac
     2461 agatggggtt tttctgtgca accaatttta agggttttca atactttaaa acacatacat
     2521 accaacactt caacgcacct ttcagcaact aaaataaaaa tgacgttatt tctatatgta
     2581 tcaagaatag aaagaactcg tttttcgcta cgctcaaaac gcaaaaaaag cactcattcg
     2641 agtgcttttt cttatcgctc caaatcatgc gattttttcc tctttgcttt tctttgctca
     2701 cgaagttctc gatcacgctg caaaacatct tgaagcgaaa aagtattctt cttttcttcc
     2761 gatcgctcat gctgacgcac gaaaagccct ctaggcgcat aggaacaact cctaaatgca
     2821 tgtgaggggt tttctcgtcc atgtgaacag tcgcatacgc aatattttgt ttcccatact
     2881 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2941 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3001 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3061 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3121 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3181 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3241 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3301 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3361 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3421 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3481 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3541 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3601 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3661 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3721 ttctacgggg gcgcgcccag ctgtctaggg cggcggattt gtcctactca ggagagcgtt
     3781 caccgacaaa caacagataa aacgaaaggc ccagtctttc gactgagcct ttcgttttat
     3841 ttgatgcct
//
