LOCUS       sequence-462186-        3394 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector part for BEVA; BEVA2.0 Position 2 Km/Nm antibiotic resistance
            via nptII, CmR/KmR/NmR..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3394)
  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 3394)
  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: pNDGG001
FEATURES             Location/Qualifiers
     source          1..3394
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        38..56
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             360..1154
                     /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(414..433)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1024..1043
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     promoter        complement(1324..1342)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(1453..2112)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2027..2046
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2113..2215)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      2316..2904
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2805..2824
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3058..3075
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3334..3352
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3334..3351
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ggagacactt gtgcctgaac accatatcca tccggcgtaa tacgactcac tatagggaga
       61 gcggccgcca gatcttccgg atggctcgag tttttcagca agataaacgt ctcagcaata
      121 tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt agcttgcagt gggcttacat
      181 ggcgatagct agactgggcg gttttatgga cagcaagcga accggaattg ccagctgggg
      241 cgccctctgg taaggttggg aagccctgca aagtaaactg gatggctttc ttgccgccaa
      301 ggatctgatg gcgcagggga tcaagatctg atcaagagac aggatgagga tcgtttcgca
      361 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
      421 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
      481 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
      541 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
      601 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
      661 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
      721 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
      781 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
      841 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
      901 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
      961 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     1021 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     1081 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     1141 acgagttctt ctgaactatg agacgtttat ctttctagaa gatctcctac aatattctca
     1201 gctgccatgg aaaatcgatg ttcttctttt attctctcaa gattttcagg ctgtatatta
     1261 accattcctg tagacttctt aattaagacg tcagaattct cgaggcggcc gcatgtgagt
     1321 ctccctatag tgagtcgtat taatttcgcg ggcggaaccc ctatttgttt atttttctaa
     1381 atacattcaa atatgtatcc gctcatgagt agcaccaggc gtttaagggc accaataact
     1441 gccttaaaaa aattacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag
     1501 cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat
     1561 cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt
     1621 gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgagac
     1681 aaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc
     1741 cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag
     1801 cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca
     1861 tatcaccagc tcaccgtctt tcattgccat acgaaattcc ggatgagcat tcatcaggcg
     1921 ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa
     1981 aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa
     2041 tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat
     2101 ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc
     2161 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaaagtc
     2221 tcattttcgc caaaagttgt catgaccaaa atcccttaac gtgagttttc gttccactga
     2281 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     2341 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2401 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     2461 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     2521 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2581 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2641 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2701 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2761 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     2821 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     2881 tcaggggggc ggagcctatg gaaaaacgcc agcaatgcgg cctttttacg gttcctggcc
     2941 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     3001 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     3061 gagtcagtga gcgaggaagc cgaaaaatca ataatcagac aacaagatgt gcgaactcga
     3121 tattttacac gactctcttt accaattctg ccccgaatta cacttaaaac gactcaacag
     3181 cttaacgttg gcttgccacg cattacttga ctgtaaaact ctcactctta ccgaacttgg
     3241 ccgtaacctg ccaaccaaag cgagaacaaa acataacatc aaacgaatcg accgattgtt
     3301 aggtaatcgt cacctgcagg aaggtttaaa cgcatttagg tgacactata gaagtgtgta
     3361 tcgctcgagg gatccgaatt cgaagtcttg gtac
//
