LOCUS       sequence-462187-        4047 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector part for BEVA; BEVA2.0 Position 2 Sp antibiotic resistance 
            via aadA, AmpR/SpR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4047)
  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 4047)
  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: pNDGG002
FEATURES             Location/Qualifiers
     source          1..4047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             756..1547
                     /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"
     primer_bind     complement(1883..1899)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1883..1899)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1896..1918)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(2071..2659)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2151..2170)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2776..2800)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2900..3691,3692..3760))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3523..3542
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3761..3865)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3964..3986
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3978..3995
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3979..3995
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cactctgtgc gtctcagcaa ccggatcaat tcccctgctc gcgcaggctg ggtgccaagc
       61 tctcgggtaa catcaaggcc cgatccttgg agcccttgcc ctcccgcacg atgatcgtgc
      121 cgtgatcgaa atccagatcc ttgacccgca gttgcaaacc ctcactgatc cgcatgcccg
      181 ttccatacag aagctgggcg aacaaacgat gctcgccttc cagaaaaccg aggatgcgaa
      241 ccacttcatc cggggtcagc accaccggca agcgccgcga cggccgaggt cttgcgatct
      301 cctgaagcca gggcagatcc gtgcacagca ccttgccgta gaagaacagc aaggccgcca
      361 atgcctgacg atgcgtggag agcgaaacct tgcgctcgtt cgccagccag gacagaaatg
      421 cctcgacttc gctgctgccc aaggttgccg ggtgacgcac accgtggaaa cggatgaagg
      481 cacgaaccca gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc
      541 gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg
      601 ttttcatggc ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc
      661 agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc
      721 agcagggcag tcgccctaaa acaaagttaa acatcatgag ggaagcggtg atcgccgaag
      781 tatcgactca actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc
      841 tggccgtaca tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg
      901 atttgctggt tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg
      961 accttttgga aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca
     1021 ccattgttgt gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat
     1081 ttggagaatg gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca
     1141 ttgatctggc tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag
     1201 cggcggagga actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa
     1261 ccttaacgct atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta
     1321 cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg
     1381 ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac
     1441 aggcttatct tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat
     1501 ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg caaataatgt ctaacaattc
     1561 gttcaagccg acgccgcttc gcggcgcggc ttaactcaag cgttagatgc actaagcaca
     1621 taattgctca cagccaaact atcaggtcaa gtctgctttt attattttta agcgtgcata
     1681 ataagcccta cacaaattgg gagatatatc atgaaaggct ggctttttct tgttatcgca
     1741 atagttggcg aagtaatcgc aacatccgca ttaaaatcta gcgagggctt tactaagctg
     1801 atccggacta tgagacgcac gaagtgaggc taggtggagg ctcagtgatg ataagtctgc
     1861 gatggtggat gcatgtgtca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     1921 gagggcacaa tcctattccg cgctatccga caatctccaa gacattaggt ggagttcagt
     1981 tcggcgtatg gcatatgtcg ctggaaagaa catgtgagca aaaggccagc aaaaggccag
     2041 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2101 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2161 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2221 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2281 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2341 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2401 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2461 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     2521 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     2581 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2641 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctctatt
     2701 caacaaagcc gccgtcccgt caagtcagcg taaatgggta gggggcttca aatcgtcctc
     2761 gtgataccaa ttcggagcct gcttttttgt acaaacttgt tgataatggc aattcaagga
     2821 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     2881 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     2941 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     3001 agggcttacc atctggcccc agtgctgcaa tgataccgcg agagccacgc tcaccggctc
     3061 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     3121 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     3181 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     3241 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     3301 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     3361 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     3421 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     3481 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     3541 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     3601 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     3661 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     3721 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     3781 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     3841 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccagat acctgaaaca
     3901 aaacccatcg tacggccaag gaagtctcca ataactgtga tccaccacaa gcgccagggt
     3961 tttcccagtc acgacgttgt aaaacgacgg ccagtcatgc ataatccgca cgcatctgga
     4021 ataaggaagt gccattccgc ctgacct
//
