LOCUS       sequence-471982-        6753 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BEVA Golden Gate cloning vector; BEVA2.0 Broad host range plasmid 
            with stability. L1-RK2-Km-par-ELT4; GG assembly from pOGG004, 
            pNDGG001, pOGG010, pOGG012, and pOGG014, NmR/KmR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6753)
  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 6753)
  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: pNDGG054
FEATURES             Location/Qualifiers
     source          1..6753
                     /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             3478..4272
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3532..3551)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4142..4161
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     oriT            4412..4520
                     /note="incP origin of transfer"
     CDS             complement(4581..5729)
                     /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      6122..6653
                     /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 aggcaatatc
     3241 tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg
     3301 cgatagctag actgggcggt tttatggaca gcaagcgaac cggaattgcc agctggggcg
     3361 ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctt gccgccaagg
     3421 atctgatggc gcaggggatc aagatctgat caagagacag gatgaggatc gtttcgcatg
     3481 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3541 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3601 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     3661 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3721 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3781 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     3841 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     3901 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     3961 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4021 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4081 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4141 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4201 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4261 gagttcttct gaactacttt tccgctgcat aaccctgctt cggggtcatt atagcgattt
     4321 tttcggtata tccatccttt ttcgcacgat atacaggatt ttgccaaagg gttcgtgtag
     4381 actttccttg gtgtatccaa cggcgtcagc cgggcaggat aggtgaagta ggcccacccg
     4441 cgagcgggtg ttccttcttc actgtccctt attcgcacct ggcggtgctc aacgggaatc
     4501 ctgctctgcg aggctggccg taggccggcc gcgatgcagg tggctgctga acccccagcc
     4561 ggaactgacc ccacaaggcc ctagcgtttg caatgcacca ggtcatcatt gacccaggcg
     4621 tgttccacca ggccgctgcc tcgcaactct tcgcaggctt cgccgacctg ctcgcgccac
     4681 ttcttcacgc gggtggaatc cgatccgcac atgaggcgga aggtttccag cttgagcggg
     4741 tacggctccc ggtgcgagct gaaatagtcg aacatccgtc gggccgtcgg cgacagcttg
     4801 cggtacttct cccatatgaa tttcgtgtag tggtcgccag caaacagcac gacgatttcc
     4861 tcgtcgatca ggacctggca acgggacgtt ttcttgccac ggtccaggac gcggaagcgg
     4921 tgcagcagcg acaccgattc caggtgccca acgcggtcgg acgtgaagcc catcgccgtc
     4981 gcctgtaggc gcgacaggca ttcctcggcc ttcgtgtaat accggccatt gatcgaccag
     5041 cccaggtcct ggcaaagctc gtagaacgtg aaggtgatcg gctcgccgat aggggtgcgc
     5101 ttcgcgtact ccaacacctg ctgccacacc agttcgtcat cgtcggcccg cagctcgacg
     5161 ccggtgtagg tgatcttcac gtccttgttg acgtggaaaa tgaccttgtt ttgcagcgcc
     5221 tcgcgcggga ttttcttgtt gcgcgtggtg aacagggcag agcgggccgt gtcgtttggc
     5281 atcgctcgca tcgtgtccgg ccacggcgca atatcgaaca aggaaagctg catttccttg
     5341 atctgctgct tcgtgtgttt cagcaacgcg gcctgcttgg cttcgctgac ctgttttgcc
     5401 aggtcctcgc cggcggtttt tcgcttcttg gtcgtcatag ttcctcgcgt gtcgatggtc
     5461 atcgacttcg ccaaacctgc cgcctcctgt tcgagccgac gcgaacgctc cacggcggcc
     5521 gatggcgcgg gcagggcagg gggagccagt tgcacgctgt cgcgctcgat cttggccgta
     5581 gcttgctgga ctatcgagcc gacggactgg aaggtttcgc ggggcgcacg catgacggtg
     5641 cggcttgcga tggtttcggc atcctcggcg gaaaaccccg cgtcgatcag ttcttgcctg
     5701 tatgccttcc ggtcaaacgt ccgattcatt caccctcctt gcgggattgc cccggaatta
     5761 attccccgga tcgatccgtc gatcttgatc ccctgcgcca tcagatcctt ggcggcaaga
     5821 aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca
     5881 attccggttc gcttgctgtc cataaaaccg cccagtctag ctatcgccat gtaagcccac
     5941 tgcaagctac ctgctttctc tttgcgcttg cgttttccct tgtccagata gcccagtagc
     6001 tgacattcat ccggggtcag caccgtttct gcggactggc tttctacgtg gctgccattt
     6061 ttggggtgag gccgttcgcg gccgaggggc gcagcccctg gggggatggg aggcccgcgt
     6121 tagcgggccg ggagggttcg agaagggggg gcacccccct tcggcgtgcg cggtcacgcg
     6181 cacagggcgc agccctggtt aaaaacaagg tttataaata ttggtttaaa agcaggttaa
     6241 aagacaggtt agcggtggcc gaaaaacggg cggaaaccct tgcaaatgct ggattttctg
     6301 cctgtggaca gcccctcaaa tgtcaatagg tgcgcccctc atctgtcagc actctgcccc
     6361 tcaagtgtca aggatcgcgc ccctcatctg tcagtagtcg cgcccctcaa gtgtcaatac
     6421 cgcagggcac ttatccccag gcttgtccac atcatctgtg ggaaactcgc gtaaaatcag
     6481 gcgttttcgc cgatttgcga ggctggccag ctccacgtcg ccggccgaaa tcgagcctgc
     6541 ccctcatctg tcaacgccgc gccgggtgag tcggcccctc aagtgtcaac gtccgcccct
     6601 catctgtcag tgagggccaa gttttccgcg aggtatccac aacgccggcg gccctacatg
     6661 gctctgctgt agtgagtggg ttgcgctccg gcagcggtcc tgatcccccg cagaaaaaaa
     6721 ggatctcaag aagatccttt gatcttttct aca
//
