LOCUS       sequence-472122-        2461 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BEVA Golden Gate cloning vector; Narrow host range plasmid with FRT 
            site. L1-pMB1-Gm-FRT-ELT4; GG assembly from pNDGG021, pOGG009, 
            pNDGG007, pNDGG009, pOGG014. GmR..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2461)
  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 2461)
  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: pNDGG012
FEATURES             Location/Qualifiers
     source          1..2461
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(41..46,47..63,64..69)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             258..791
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     oriT            985..1093
                     /note="incP origin of transfer"
     rep_origin      complement(1163..1751)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1243..1262)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1832..1879
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     protein_bind    1964..1985
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2000..2005,2006..2023,2024..2030)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2038..2054
                     /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     2043..2065
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2062..2078
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2062..2078
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2074..2397
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    2091..2147
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2148..2165)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2148..2164)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2157..2179)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2373..2392)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 catccgctta cagacaagct gtgacggtct cacgctgcaa ttgacataag cctgttcggt
       61 tcgtaaactg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga
      121 acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgt
      181 acagcctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat
      241 gttatggagc agcaacgatg ttacgcagca gcaacgatgt tacgcagcag ggcagtcgcc
      301 ctaaaacaaa gttaggtggc tcaagtatgg gcatcattcg cacatgtagg ctcggccctg
      361 accaagtcaa atccatgcgg gctgctcttg atcttttcgg tcgtgagttc ggcgacgtag
      421 ccacctactc ccaacatcag ccggactccg attacctcgg gaacttgctc cgtagtaaga
      481 cattcatcgc gcttgctgcc ttcgaccaag aagcggttgt tggcgctctc gcggcttacg
      541 ttctgcccaa gtttgagcag ccgcgtagtg agatctatat ctatgatctc gcagtctccg
      601 gagagcaccg gaggcagggc attgccaccg cgctcatcaa tctcctcaag catgaggcca
      661 acgcgcttgg tgcttatgtg atctacgtgc aagcagatta cggtgacgat cccgcagtgg
      721 ctctctatac aaagttgggc atacgggaag aagtgatgca ctttgatatc gacccaagta
      781 ccgccaccta acaattcgtt caagccgaga tcggcttccc ggccgcggag ttgttcggta
      841 aattgactac ttttccgctg cataaccctg cttcggggtc attatagcga ttttttcggt
      901 atatccatcc tttttcgcac gatatacagg attttgccaa agggttcgtg tagactttcc
      961 ttggtgtatc caacggcgtc agccgggcag gataggtgaa gtaggcccac ccgcgagcgg
     1021 gtgttccttc ttcactgtcc cttattcgca cctggcggtg ctcaacggga atcctgctct
     1081 gcgaggctgg ccgataagct agcttgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1141 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1201 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1261 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1321 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     1381 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     1441 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     1501 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     1561 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     1621 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     1681 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     1741 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     1801 actcacgtta agggattttg gtcatgatta cgaagttcct attccgaagt tcctattctc
     1861 tagaaagtat aggaacttca gggatccgtc gaccagagga gtgagaccgc agctggcacg
     1921 acaggtttgc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     1981 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     2041 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcttgcat
     2101 gcctgcaggt cgactctaga ggatccccgg gtaccgagct cgaattcact ggccgtcgtt
     2161 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     2221 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     2281 ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc
     2341 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     2401 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     2461 g
//
