LOCUS       sequence-466017-        6522 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BEVA Golden Gate cloning vector; BEVA2.0 Broad host range plasmid 
            with stability. L1-RK2-Gm-par-ELT4; GG assembly from pOGG004, 
            pOGG009, pOGG010, pOGG012, and pOGG014, GmR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6522)
  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 6522)
  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: pNDGG050
FEATURES             Location/Qualifiers
     source          1..6522
                     /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"
     promoter        join(3237..3242,3243..3259,3260..3265)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3454..3987
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     oriT            4181..4289
                     /note="incP origin of transfer"
     CDS             complement(4350..5498)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      5891..6422
                     /label=oriV
                     /note="incP origin of 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 aggcaattga
     3241 cataagcctg ttcggttcgt aaactgtaat gcaagtagcg tatgcgctca cgcaactggt
     3301 ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt
     3361 atgactgttt ttttgtacag cctatgcctc gggcatccaa gcagcaagcg cgttacgccg
     3421 tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagcaa cgatgttacg
     3481 cagcagggca gtcgccctaa aacaaagtta ggtggctcaa gtatgggcat cattcgcaca
     3541 tgtaggctcg gccctgacca agtcaaatcc atgcgggctg ctcttgatct tttcggtcgt
     3601 gagttcggcg acgtagccac ctactcccaa catcagccgg actccgatta cctcgggaac
     3661 ttgctccgta gtaagacatt catcgcgctt gctgccttcg accaagaagc ggttgttggc
     3721 gctctcgcgg cttacgttct gcccaagttt gagcagccgc gtagtgagat ctatatctat
     3781 gatctcgcag tctccggaga gcaccggagg cagggcattg ccaccgcgct catcaatctc
     3841 ctcaagcatg aggccaacgc gcttggtgct tatgtgatct acgtgcaagc agattacggt
     3901 gacgatcccg cagtggctct ctatacaaag ttgggcatac gggaagaagt gatgcacttt
     3961 gatatcgacc caagtaccgc cacctaacaa ttcgttcaag ccgagatcgg cttcccggcc
     4021 gcggagttgt tcggtaaatt gactactttt ccgctgcata accctgcttc ggggtcatta
     4081 tagcgatttt ttcggtatat ccatcctttt tcgcacgata tacaggattt tgccaaaggg
     4141 ttcgtgtaga ctttccttgg tgtatccaac ggcgtcagcc gggcaggata ggtgaagtag
     4201 gcccacccgc gagcgggtgt tccttcttca ctgtccctta ttcgcacctg gcggtgctca
     4261 acgggaatcc tgctctgcga ggctggccgt aggccggccg cgatgcaggt ggctgctgaa
     4321 cccccagccg gaactgaccc cacaaggccc tagcgtttgc aatgcaccag gtcatcattg
     4381 acccaggcgt gttccaccag gccgctgcct cgcaactctt cgcaggcttc gccgacctgc
     4441 tcgcgccact tcttcacgcg ggtggaatcc gatccgcaca tgaggcggaa ggtttccagc
     4501 ttgagcgggt acggctcccg gtgcgagctg aaatagtcga acatccgtcg ggccgtcggc
     4561 gacagcttgc ggtacttctc ccatatgaat ttcgtgtagt ggtcgccagc aaacagcacg
     4621 acgatttcct cgtcgatcag gacctggcaa cgggacgttt tcttgccacg gtccaggacg
     4681 cggaagcggt gcagcagcga caccgattcc aggtgcccaa cgcggtcgga cgtgaagccc
     4741 atcgccgtcg cctgtaggcg cgacaggcat tcctcggcct tcgtgtaata ccggccattg
     4801 atcgaccagc ccaggtcctg gcaaagctcg tagaacgtga aggtgatcgg ctcgccgata
     4861 ggggtgcgct tcgcgtactc caacacctgc tgccacacca gttcgtcatc gtcggcccgc
     4921 agctcgacgc cggtgtaggt gatcttcacg tccttgttga cgtggaaaat gaccttgttt
     4981 tgcagcgcct cgcgcgggat tttcttgttg cgcgtggtga acagggcaga gcgggccgtg
     5041 tcgtttggca tcgctcgcat cgtgtccggc cacggcgcaa tatcgaacaa ggaaagctgc
     5101 atttccttga tctgctgctt cgtgtgtttc agcaacgcgg cctgcttggc ttcgctgacc
     5161 tgttttgcca ggtcctcgcc ggcggttttt cgcttcttgg tcgtcatagt tcctcgcgtg
     5221 tcgatggtca tcgacttcgc caaacctgcc gcctcctgtt cgagccgacg cgaacgctcc
     5281 acggcggccg atggcgcggg cagggcaggg ggagccagtt gcacgctgtc gcgctcgatc
     5341 ttggccgtag cttgctggac tatcgagccg acggactgga aggtttcgcg gggcgcacgc
     5401 atgacggtgc ggcttgcgat ggtttcggca tcctcggcgg aaaaccccgc gtcgatcagt
     5461 tcttgcctgt atgccttccg gtcaaacgtc cgattcattc accctccttg cgggattgcc
     5521 ccggaattaa ttccccggat cgatccgtcg atcttgatcc cctgcgccat cagatccttg
     5581 gcggcaagaa agccatccag tttactttgc agggcttccc aaccttacca gagggcgccc
     5641 cagctggcaa ttccggttcg cttgctgtcc ataaaaccgc ccagtctagc tatcgccatg
     5701 taagcccact gcaagctacc tgctttctct ttgcgcttgc gttttccctt gtccagatag
     5761 cccagtagct gacattcatc cggggtcagc accgtttctg cggactggct ttctacgtgg
     5821 ctgccatttt tggggtgagg ccgttcgcgg ccgaggggcg cagcccctgg ggggatggga
     5881 ggcccgcgtt agcgggccgg gagggttcga gaaggggggg cacccccctt cggcgtgcgc
     5941 ggtcacgcgc acagggcgca gccctggtta aaaacaaggt ttataaatat tggtttaaaa
     6001 gcaggttaaa agacaggtta gcggtggccg aaaaacgggc ggaaaccctt gcaaatgctg
     6061 gattttctgc ctgtggacag cccctcaaat gtcaataggt gcgcccctca tctgtcagca
     6121 ctctgcccct caagtgtcaa ggatcgcgcc cctcatctgt cagtagtcgc gcccctcaag
     6181 tgtcaatacc gcagggcact tatccccagg cttgtccaca tcatctgtgg gaaactcgcg
     6241 taaaatcagg cgttttcgcc gatttgcgag gctggccagc tccacgtcgc cggccgaaat
     6301 cgagcctgcc cctcatctgt caacgccgcg ccgggtgagt cggcccctca agtgtcaacg
     6361 tccgcccctc atctgtcagt gagggccaag ttttccgcga ggtatccaca acgccggcgg
     6421 ccctacatgg ctctgctgta gtgagtgggt tgcgctccgg cagcggtcct gatcccccgc
     6481 agaaaaaaag gatctcaaga agatcctttg atcttttcta ca
//
