LOCUS       sequence-472119-        8431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BEVA Golden Gate cloning vector; BEVA2.0 Narrow host range plasmid 
            with I-SceI counterselectable site for BpiI cloning; KmR/NmR..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8431)
  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 8431)
  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: pQGG003
FEATURES             Location/Qualifiers
     source          1..8431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(222..316)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     regulatory      4402..4411
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      5948..6034
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(6298..6315)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6432..6977)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(6512..6531)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            7360..7469
                     /note="incP origin of transfer"
     CDS             complement(7609..8403)
                     /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(7720..7739)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     8330..8349
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tcagatcttg atcccctgcg ccatcagatc cttggcggca agaaagccat ccagtttact
       61 ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg ttcgcttgct
      121 gtccataaaa ccgcccagtc tagctatcgc catgtaagcc cactgcaagc tacctgcttt
      181 ctctttgcgc ttgcgttttc ccttgtccag atattgcctg gattctcacc aataaaaaac
      241 gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga
      301 tctatcaaca ggagtccaag tcccttgtct tcgtgctaat cgccagcgta tgcatcacgc
      361 cgtcaggggc aaagaaggtt gtgtttcttt tggcttcctc tatgcgccgc ccctgtcaaa
      421 acttcaggcg acgctccggg aaagacatgg cgctagagcg ggcgctgcca gagatgcgca
      481 gggcggggag gatgagcccg catccgggaa gtaagggcct gaccagaggc ggccagcagc
      541 agcgttaatt tttcgggcgt ggtcgttgac tgccgctgat cccaggcttg ctgaccggcc
      601 tgttcaactt tgacacctat tttccggtaa acctgcttcg ccgtagcgat tgcccaggcg
      661 gaacgcaggg gcaaccctgc caggccggct gtggcagaca aatagtaagg ttctgcttcc
      721 tgcaccaaac gacgggcgat acggctcagc gcctgacggt tttcaggtgc cgcataattc
      781 tctttgttca gaccttcatg ctccagccag cttgccggca gataacagcg gcccgcatgc
      841 gcatcgtcca caatatcgcg agcaatattg gtcaactgaa atgccagccc aaggtcacag
      901 gcgcggtcca gcgtggcgtt atcccgcacg cccatgattt gcgccatcat caagccgaca
      961 acgcctgcaa cgtgatagca atagcgcagc gtatcatcca gttggctgta ttgcgcttcg
     1021 cgtacatcca tggcgaagcc ttccagatga tcaaacgcgt aagccggggc gatatcatga
     1081 gccatagcca cttcctgaaa agccgcaaac gccggttcgt gcatctgcga tcctgcatag
     1141 gcctggcgcg ttttcatctc aagttgcatc agacgttgtt cgggcgtttg taaggcaggc
     1201 tgccgggcct gaaagcccag cgtctgatcg tcaataacat cgtcacaatg gcggcaccag
     1261 gcgtagagca tcagtacgct gcgccgggtt tttgcatcaa ataactttga ggctgtcgca
     1321 aaacttttcg agccaactgc catcgtttcg accgcatgat tgagtaacga cggattattc
     1381 aaatcagatc ctccagcatc aaacctgctg tcgcttttgc cgagccgatg acgccaggaa
     1441 tgcctgcgcc gggatgcgtg cctgcgccga ccaggtagag attagtaatg gttttatcgc
     1501 ggttatgcgg ccgaaaccag gcgctctggg taagaacggg ctccacagaa aaggctgagc
     1561 catgataggc attaagctgg tcgcgaaaat caaacggcgt aaacatccgg tgcgtgacca
     1621 gctgactccg taagccaggc atgtaatgct gctcaaggta cgcaaaaata cggtcgcgta
     1681 gttttggccc ctcaaccgtc cagtcgaggt tcgcggtgcc taaatgcggc accggcgcca
     1741 acacatagta actgccgcaa ccttcaggcg ccagtgacga atccgtgaca cagggcgcgt
     1801 gcagataaag tgagaagtcc tctgcgaggc catcatgatt aaaaatttcg tcaatcagct
     1861 cgcggtaacg cgggccgaaa caaaccgtgt gatgcgcgag ctgatcatga tggtgattca
     1921 aaccaaaata gagcacaaac agagagttac tcatgcgctt agtctgcagt ttgttggact
     1981 gcttaaccgc ggcagggtgc tggcttaaca ggtcgcgata ggtatgaacc acatctgcat
     2041 ttgacgcgac ggcttgcgtc aggaacctgc gaccgtcctc taaatgcacg gcttcaatct
     2101 tgtttcctgt cgtttccata tggctgactc tggcgtttaa cacgacttcg ccacccagat
     2161 cctgaaacag ctttatcatc ccctgaacta atgcgccggt gccgccacgc ggaaaccaga
     2221 cgccccactc acgctccagc gcgtgtatca acgtataaat ggatgaggtg gcgaagggat
     2281 tgccgcccac caacagcgag tggaaagaaa acgcctggcg cagatgttca tcttcgatgt
     2341 aactggcaac cttactgtaa acgcttctcc atgcctgcag tttcgccagt tgaggtgcgg
     2401 cgcgaagcat gtctctgaac gataaaaaag ggacagtacc gagctttaga tagccttctt
     2461 taaacaccgc gcgtgaatag tccagaaact gacgataacc ttcgacatcg cggggattaa
     2521 actgctgaat ctgcgcttcg agccgggttt gatcgttatc gtaattaaag accttccctg
     2581 actcccaaca caggcggtaa aacggcgtaa ccggcagcag ttcgacatac tcttttaact
     2641 gttttcctgc cagtgcaaac agttcttcaa tggcactggg atcggtgata accgtcgggc
     2701 ctgcatcaaa ggtaaacccc tgatcctcgt agacataagc ccgaccgccg ggtttatcac
     2761 gttgttcaag cagtaagacg gggatccccg cagcttgtag acgaattgcc agtgccaggc
     2821 caccgaagcc tgcaccaatt accgtagttg gtttcatgta gtcgctcttt aacgatgagt
     2881 cgtcataatg gcttgcaatg ctgctaatac cggaacaggc ggcttgccgc tcagaatacg
     2941 tagccgatcg gtcagcgtga gttttcccgc ataaaaacgg gcaattaaat cttcaggtaa
     3001 accataaaaa cgctgcataa cccgccagcg tgaatcggcg ggtccggcta aaaacagcat
     3061 gcgattcagc atgcggaaaa agccctgctg ctgccagcgc tcgcgggcaa aatgcgtaat
     3121 ggcatggtga attgaggccg acgtaaagac atcaagtgca ctcaggcggt cggccacggc
     3181 aaccgccagc ggcagtgaat agccggtggt aggatggaac agaccggcac gtaatccact
     3241 acaggccagg gggcgctgct gccagaatgc gtcggcattg cccgacagag taatgggtaa
     3301 ggcgccctgt tcttctcgca gcagtgtctg aagctgccaa ccctgttgcg cggcatagtc
     3361 gcaaatattt tgccgcgcgc attcaggatc taatgtcgca ttatcaatat agtgcgtatc
     3421 ttcaattaac aatctggtcg gcgagagcgg caggctgtac acgaagcgat aaccattttg
     3481 ctgatcgacc gtggcatcca tgataatggg agatgataaa ccatgcgggt ggctcaatcg
     3541 ccattcctgg ccaataaacg cctggaagcc cacgctcagt gctgaatttg ccgcataacc
     3601 ccgcccgtca atcaccgcgc gggcaccgat aacctgaccc tttttcaacc gaacagattc
     3661 cgcattaacc tctgcgaccg cggtatccat ccacaagtgc gggccaaact gtcgctgtaa
     3721 aacctcagcg aaacgctgag aagtaataca aaagtagccg ctgttcagct tacgacggcg
     3781 tgtgggaaag cgtacctgat agtcgggcca gtgatgaacc accagcggag ctatccaacg
     3841 atgttggctc tcagtcaaat catcgtggtg aaatgaccac gtatgattcc cgcccgcctg
     3901 gggtgcggcg tcgataagca aaatacgcat atcaggttgc tgctgctgaa gtcgcagggc
     3961 gataaggcca ttcgcgagtc cagcccccac gagaatcaga tcataatgcg gttgcatagc
     4021 cgctcccact taagtcactc atgcggtgtc ccccttggtg cgggtgctgg gcaggcgcca
     4081 ccacggcacc gtcgggtgca ggtggtgttc gtgatgataa ccgccaaagt gaaaacaggt
     4141 cagcagcgac acggggtcgc tgatccgcga cgaccgcgca ttgtggcggt ccgggaacgc
     4201 gtcgtggccg gggcggtgcg gcagccaggt gccgaacacg aacagctgga tcgacgccag
     4261 gatcgacggc agcggccaga ataccacgta catccagcga tccccaagga tcagcgcata
     4321 gaccgtcacg atgacgggca gaagcagccc ctcgcgccag ccgaaatagg tgccgatgaa
     4381 gcgggcgtac cagcggaccg ggccgccatg gtcgaaatcg gggtcgtcgt cggttccggc
     4441 atggcggtga tgggccatgt gcttgacgat catcttgcgc cacgaaaatc cggcatacag
     4501 ccacaggaca agctggccca tcgccgcatt ggcgcgcgga cgccccggca ccaccgaccc
     4561 gtgcatcgcg tcatgcgcga tgatgaacaa tccgaccgac agccaggtca gccccaggaa
     4621 atttgcgatc gccaggatgg gatgcgccgc tgcgtccaga aaccacagcg catgcacatg
     4681 cagggccagc caagcggcga tgatgccgcc cgatacgatc aggctggtgg cggtcagatc
     4741 tgccttgggc agggcatgtg cgctcatgca gcatccttaa ctgacggcag cgagtttttt
     4801 gtcaaaccag gcctgaataa aatgttgagt ggcgtgcccg tgttggcagg ccgcagagag
     4861 atgctcactg gcaagctgaa gatgttgtct cagacgttct tcaaccgccc tcgggcctaa
     4921 cagattgacc agcgtcgatt taccggcgtc ctgattgcta tccttaccgg tgtcggtcat
     4981 gccatcggtc aaatcgtcca gcagttgaaa tgcctgacca agatcaagtg aaaaacgatg
     5041 caggcaatca cgcgcttcgc tggaggcatt cgcaacaatc gaggccatct gcatggaggc
     5101 acaaaacagc gtgctggttt taaagtgatt cgtcatcaaa atagcttcag cgctgcgcgg
     5161 cttatcccct tcagacagat ccttgaactg accctgaacc aatccttgca tgccgatggc
     5221 gtttgacagt tcagaaaccg cccgattttt tgccagcggc gtgaggccat ctgcatcggc
     5281 aattacgcca aaggctttac tcagcaaggc aaccgccgcc agtattgcca catgctctcc
     5341 gtaatgagaa tgaatggtag ggcgtccgcg ccgcagcttc gcatcgtcca tgcagggcat
     5401 atcgtcaagg atcagcgaag ccgcgtggac catttccacc gcacaggcca aatccagtaa
     5461 tccgtcatgg ctgacagcgc aacccagatc gcgggcggtc agcaacagca acatggggcg
     5521 aatacgtttt cccggtgcca gcgcaccttc acgcatcgcg gcacccacaa catcccgttc
     5581 tccctccacg ggcaataact gatcaaggcg tcgatcaata tcagccagta actgctccgc
     5641 agcatcgcga gtgagatgaa cgtgtttttt tgcgcagacc gtcattcggg ctgtccttat
     5701 aaacggatac atttacggca acgcgaaaga tgcggtgccc ttcaaaaaga cgcttaatct
     5761 gccataacat ggttggcagg cttacccgtt attttacatt tagggttaca ggtttttttt
     5821 gctaaatcca catcgttaat actagttgag cttgtggaaa aggtggaaaa ttcataaacc
     5881 tccgtcggat tggcagaccg tgcggtaccc tccgtcgagt cgaccagaag acaaggcagg
     5941 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     6001 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ccctagatct ggactacggt
     6061 tccaaattac cctgttatcc ctaccttgga atggtcagta actgtccctc ctgttcagct
     6121 actgacgggg tggtgcgtaa cggcaaaagc accgccggac atcagcgcta gcggagtgta
     6181 tactggctta ctatgttggc actgatgagg gtgtcagtga agtgcttcat gtggcaggag
     6241 aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac aggatatatt ccgcttcctc
     6301 gctcactgac tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg
     6361 gggcggagat ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg
     6421 gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca
     6481 aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc
     6541 tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg
     6601 ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct
     6661 ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg
     6721 tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg
     6781 atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt
     6841 ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa
     6901 ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca
     6961 gaccaaaacg atctcaagaa gatcatctta ttaatcagat aaaatatttc tagctagagt
     7021 cgatcttcgc cagcagggcg aggatcgtgg catcaccgaa ccgcgccgtg cgcgggtcgt
     7081 cggtgagcca gagtttcagc aggccgccca ggcggcccag gtcgccattg atgcgggcca
     7141 gctcgcggac gtgctcatag tccacgacgc ccgtgatttt gtagccctgg ccgacggcca
     7201 gcaggtaggc cgacaggctc atgccggccg ccgccgcctt ttcctcaatc gctcttcgtt
     7261 cgtctggaag gcagtacacc ttgataggtg ggctgccctt cctggttggc ttggtttcat
     7321 cagccatccg cttgccctca tctgttacgc cggcggtagc cggccagcct cgcagagcag
     7381 gattcccgtt gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa cacccgctcg
     7441 cgggtgggcc tacttcacct atcctgcccg gctgacgccg ttggatacac caaggaaagt
     7501 ctacacgaac cctttggcaa aatcctgtat atcgtgcgaa aaaggatgga tataccgaaa
     7561 aaatcgctat aatgaccccg aagcagggtt atgcagcgga aaagtagttc agaagaactc
     7621 gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac
     7681 gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc
     7741 tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg
     7801 gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc
     7861 gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg
     7921 ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc
     7981 gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg
     8041 ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag
     8101 atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc
     8161 gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc
     8221 ctgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg
     8281 cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata
     8341 gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat
     8401 catgcgaaac gatcctcatc ctgtctcttg a
//
