LOCUS       sequence-463765-        6802 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BEVA Golden Gate cloning vector; BEVA2.0 Broad host range plasmid 
            with stability. L1-pBBR1-Sp-par-ELT4; GG assembly from pOGG004, 
            pNDGG002, pOGG011, pOGG012, and pOGG014, SpR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6802)
  AUTHORS   Geddes BA, Williamson R, Schumacher J, Ardi A, Levin G, Cervenka E, 
            Huang R, diCenzo G
  TITLE     BEVA2.0: modular assembly of golden gate-compatible vectors with 
            expanded utility for genetic engineering.
  JOURNAL   Can J Microbiol. 2025 Jan 1;71:1-13. doi: 10.1139/cjm-2024-0246.
  PUBMED    39919271
REFERENCE   2  (bases 1 to 6802)
  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: "Can J 
            Microbiol."; date: "2025-01-1"; volume: "71:1-13. doi"; pages: " 
            10.1139/cjm-2024-0246"
COMMENT     Sequence Label: pNDGG051
FEATURES             Location/Qualifiers
     source          1..6802
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(2422..2508)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    2596..2617
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2632..2637,2638..2655,2656..2662)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2670..2686
                     /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     2675..2697
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2694..2710
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2694..2710
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2706..3029
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    2723..2779
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2780..2797)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2780..2796)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2789..2811)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3005..3024)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      3134..3228
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3972..4763
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     oriT            5162..5270
                     /note="incP origin of transfer"
     CDS             complement(5281..5943)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      5944..6715
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
ORIGIN
        1 ttactgcatg agcttgtgga agtgtgctga gcataaatga ttctcctagc tgttctttgg
       61 gtaagtacgc catcaggacg ttgtgagtgg cgcgattttt agcggctgaa atcagccctt
      121 gagcctgtcg gcaagtcgcg tcatgaggtc catgcgctca tgcaggatcg ccacgaccaa
      181 cgcgggttcg cccgcacgcg gcaggcaaaa aacgtagtgg tgttcgcagc gggccatccg
      241 cagcgcggga aagagttcgc tcatgtcctt aaacgggcct tcgccggcgg caagcctggc
      301 tatgccctgt tccagcttag cgatatagcg gcgcacctgc gccgcgcccc actcccggcg
      361 cgtgtagcgg atgatgccgc gtagatcggc ttcggcctca gccgtgagga tgtaggccgt
      421 caagcgcgat ccccgctgag ttcttcatca agaatttcgc cgacgctctt ggtggacacc
      481 ttgccggcaa gcccatcgtt gatgcggttc cccagcatgg ttttcagttc ctgccatgcc
      541 tgatcggcat cagcgtcacc ggggaacaga cgttcgaggg cgtattgctt aatggtcttg
      601 ccctgcaagg cggccagggc tttcaggctc tggtgctgct ggtccgtcat gtcgattgtc
      661 aggcggctca ttggataacc tccataaaat acacgtaacc acattagcac atatgtgggc
      721 gtgaggctac agcgcgaggc gcattaaggt cgggaaaatg cgctaggcgc atttaaattg
      781 cgtattgctg taatgcgcca tgccggctag actaggccca aatgggtata cccaatttga
      841 ccaaggggga cgcgatgagg gcggccaagc actaccgaca acttctatcc atcgacttca
      901 acatcgaggc gctggccttc gtgcctggac ccgacggcac acgcggccgg cgcatccacg
      961 tcctggggcg cgaggtccgc gaccggcccg gcctggtcga gtacctttcg ccggcgttcg
     1021 gctcgcgggt ggcgctggac ggctactgca aggccaattt cgatgcagtg ctgcacctgg
     1081 cgtaccccga tcatcagcaa tggggccacg catgaagcgc cgaagctacg ccatgctgcg
     1141 cgccgctgcc gcgctggccg tcctggtcgt tgcctcgccg gcatgggccg agctgcgcgg
     1201 cgaggtcgtg cgcatcatcg acggcgacac catcgacgtg ctggtagaca agcagccggt
     1261 gcgcgtgcgc ctggtggaca ttgacgcgcc ggaaaagcgg caagccttcg gcgaacgtgc
     1321 gcgccaggcg ctggccggca tggtgttccg ccggcacgtc ctggtcgacg agaaggacac
     1381 cgaccgttac ggccgcacgc tgggcaccgt ctgggtcaac atggagctgg ccagccggcc
     1441 gccgcagccg cgcaacgtca acgccgcgat ggttcaccag ggcatggcgt gggcctatcg
     1501 cttccacggc cgcgcggccg accctgaaat gctgcggctc gaacaggagg cgcgaggcaa
     1561 gcgcgtcggc ctctggtccg atccgcacgc cgtcgagccg tggaaatggc gacgcgagag
     1621 caacaaccgg agggacgaag gttgaaggtc gcccgcatct acctgcgcgc cagtacggac
     1681 gagcagaatc ttgaacgcca ggagagcctt gtagcggcca cgcgggccgc cgggtactac
     1741 gtcgccggca tctaccgcga gaaggcgtcc ggcgcacgcg ccgaccggcc cgagctgctg
     1801 cgcatgatcg cggacctgca acctggtgaa gtcgtcgttg cggagaagat cgaccgcatc
     1861 agccgcttgc cgttggccga ggccgagcgc ctggttgcgt cgatccgggc caaaggggcc
     1921 aagctggccg tgcctggcgt ggtggacctg tcggagctgg ccgccgaggc gaacggagtg
     1981 gcgaaaatcg ttctggaatc cgtccaggac atgcttttga agctcgcctt gcagatggcc
     2041 cgcgacgact acgaggatcg gcgcgagcgt caacgtcagg gtgtccagtt ggcgaaggcc
     2101 gccggccgct acaccggccg caaacgtgac gccggcatgc acgaccgcat catcacgctt
     2161 cgctccggcg gatcgagcat tgccaagacg gccaagctgg tcggatgcag cccgagccag
     2221 gtcaaacgag tgtgggcggc ctggaacgcg cagcagcaaa aataaagccg ggcagtgccc
     2281 ggcttttctc accttttcgc gtcccgcagg gccgctgcga gcgccctacc tagatcctcg
     2341 ctttccccct cggtgtagtc cggccagggc acgaagggcg cggatgcgaa cctgttgagc
     2401 aggtcagatc tagggcggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
     2461 gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctgccgaa
     2521 ttcggatccg gagtgagacc gcagctggca cgacaggttt gccgactgga aagcgggcag
     2581 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     2641 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     2701 cagctatgac catgattacg ccaagcttgc atgcctgcag gtcgactcta gaggatcccc
     2761 gggtaccgag ctcgaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     2821 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     2881 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc
     2941 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
     3001 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     3061 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgacgg
     3121 tctcacgctc ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat
     3181 ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aggcaaccgg
     3241 atcaattccc ctgctcgcgc aggctgggtg ccaagctctc gggtaacatc aaggcccgat
     3301 ccttggagcc cttgccctcc cgcacgatga tcgtgccgtg atcgaaatcc agatccttga
     3361 cccgcagttg caaaccctca ctgatccgca tgcccgttcc atacagaagc tgggcgaaca
     3421 aacgatgctc gccttccaga aaaccgagga tgcgaaccac ttcatccggg gtcagcacca
     3481 ccggcaagcg ccgcgacggc cgaggtcttg cgatctcctg aagccagggc agatccgtgc
     3541 acagcacctt gccgtagaag aacagcaagg ccgccaatgc ctgacgatgc gtggagagcg
     3601 aaaccttgcg ctcgttcgcc agccaggaca gaaatgcctc gacttcgctg ctgcccaagg
     3661 ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg aacccagtgg acataagcct
     3721 gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct
     3781 tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt
     3841 tttttggggt acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc
     3901 gatgtttgat gttatggagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa
     3961 agttaaacat catgagggaa gcggtgatcg ccgaagtatc gactcaacta tcagaggtag
     4021 ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg tacggctccg
     4081 cagtggatgg cggcctgaag ccacacagtg atattgattt gctggttacg gtgaccgtaa
     4141 ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact tcggcttccc
     4201 ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac gacgacatca
     4261 ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca
     4321 ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc ttgctgacaa
     4381 aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc tttgatccgg
     4441 ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc
     4501 ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg
     4561 cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc
     4621 cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag
     4681 atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg aaaggcgaga
     4741 tcaccaaggt agtcggcaaa taatgtctaa caattcgttc aagccgacgc cgcttcgcgg
     4801 cgcggcttaa ctcaagcgtt agatgcacta agcacataat tgctcacagc caaactatca
     4861 ggtcaagtct gcttttatta tttttaagcg tgcataataa gccctacaca aattgggaga
     4921 tatatcatga aaggctggct ttttcttgtt atcgcaatag ttggcgaagt aatcgcaaca
     4981 tccgcattaa aatctagcga gggctttact aagctgatcc ggactacttt tccgctgcat
     5041 aaccctgctt cggggtcatt atagcgattt tttcggtata tccatccttt ttcgcacgat
     5101 atacaggatt ttgccaaagg gttcgtgtag actttccttg gtgtatccaa cggcgtcagc
     5161 cgggcaggat aggtgaagta ggcccacccg cgagcgggtg ttccttcttc actgtccctt
     5221 attcgcacct ggcggtgctc aacgggaatc ctgctctgcg aggctggccg taggccggcc
     5281 ctaccggcgc ggcagcgtta cccgtgtcgg cggctccaac ggctcgccat cgtccagaaa
     5341 acacggctca tcgggcatcg gcaggcgctg ctgcccgcgc cgttcccatt cctccgtttc
     5401 ggtcaaggct ggcaggtctg gttccatgcc cggaatgccg ggctggctgg gcggctcctc
     5461 gccggggccg gtcggtagtt gctgctcgcc cggatacagg gtcgggatgc ggcgcaggtc
     5521 gccatgcccc aacagcgatt cgtcctggtc gtcgtgatca accaccacgg cggcactgaa
     5581 caccgacagg cgcaactggt cgcggggctg gccccacgcc acgcggtcat tgaccacgta
     5641 ggccgacacg gtgccggggc cgttgagctt cacgacggag atccagcgct cggccaccaa
     5701 gtccttgact gcgtattgga ccgtccgcaa agaacgtccg atgagcttgg aaagagtctt
     5761 ttggctgacc accacggcgt tctggtggcc catctgcgcg acgaggtgat gcagcagcat
     5821 tgccgccgtg ggtttcctcg caataagccc ggcccacgcc tcatgcgctt tgcgttccgt
     5881 ttggacccag tgaccgggct tgttcttggc ttgaatgccg atttctctgg actgcgtggc
     5941 catgcttatc tccatgcggt aggggtgccg cacggttgcg gcaccatgcg caatcagctg
     6001 caacttttcg gcagcgcgac aacaattatg cgttgcgtaa aagtggcagt caattacaga
     6061 ttttctttaa cctacgcaat gagctattgc ggggggtgcc gcaatgagct gttgcgtacc
     6121 cccctttttt aagttgttga tttttaagtc tttcgcattt cgccctatat ctagttcttt
     6181 ggtgcccaaa gaagggcacc cctgcggggt tcccccacgc cttcggcgcg gctccccctc
     6241 cggcaaaaag tggcccctcc ggggcttgtt gatcgactgc gcggccttcg gccttgccca
     6301 aggtggcgct gcccccttgg aacccccgca ctcgccgccg tgaggctcgg ggggcaggcg
     6361 ggcgggcttc gcccttcgac tgcccccact cgcataggct tgggtcgttc caggcgcgtc
     6421 aaggccaagc cgctgcgcgg tcgctgcgcg agccttgacc cgccttcctc ttggtgtcca
     6481 accggcaagc gaagcgcgca ggccgcaggc cggaggcttt tccccagaga aaattaaaaa
     6541 aattgatggg gcaaggccgc aggccgcgca gttggagccg gtgggtatgt ggtcgaaggc
     6601 tgggtagccg gtgggcaatc cctgtggtca agctcgtggg caggcgcagc ctgtccatca
     6661 gcttgtccag cagggttgtc cacgggccga gcgaagcgag ccagccggtg gccgctcgcg
     6721 gccatcgtcc acatatccac gggctggcaa gggagcgcag cgaccgcgca gggcgaagcc
     6781 cggagagcaa gcccgtaggg gg
//
