LOCUS       sequence-489467-        3260 bp    DNA     circular SYN 16-OCT-2025
DEFINITION  BEVA Golden Gate cloning vector; Narrow host range plasmid. 
            L1-p15A-Gm-ELT3; pNDGG021, pOGG009, pNDGG006, pOGG013. NmR..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3260)
  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 3260)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 16, 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: pNDGG015
FEATURES             Location/Qualifiers
     source          1..3260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    71..92
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(107..112,113..130,131..137)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    145..161
                     /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     150..172
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     169..185
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     169..185
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             181..504
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    198..254
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(255..272)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(255..271)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(264..286)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(480..499)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             850..1644
                     /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(904..923)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1514..1533
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     oriT            1784..1893
                     /note="incP origin of transfer"
     rep_origin      2276..2821
                     /direction=RIGHT
                     /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     2722..2741
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2938..2955
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3169..3255)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tgccggagtg agaccgcagc tggcacgaca ggtttgccga ctggaaagcg ggcagtgagc
       61 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
      121 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
      181 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccgggta
      241 ccgagctcga attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
      301 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
      361 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
      421 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
      481 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
      541 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gacggtctca
      601 cgctgcaata tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt agcttgcagt
      661 gggcttacat ggcgatagct agactgggcg gttttatgga cagcaagcga accggaattg
      721 ccagctgggg cgccctctgg taaggttggg aagccctgca aagtaaactg gatggctttc
      781 ttgccgccaa ggatctgatg gcgcagggga tcaagatctg atcaagagac aggatgagga
      841 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
      901 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
      961 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     1021 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     1081 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     1141 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     1201 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     1261 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     1321 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     1381 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     1441 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     1501 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     1561 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     1621 cgccttcttg acgagttctt ctgaactact tttccgctgc ataaccctgc ttcggggtca
     1681 ttatagcgat tttttcggta tatccatcct ttttcgcacg atatacagga ttttgccaaa
     1741 gggttcgtgt agactttcct tggtgtatcc aacggcgtca gccgggcagg ataggtgaag
     1801 taggcccacc cgcgagcggg tgttccttct tcactgtccc ttattcgcac ctggcggtgc
     1861 tcaacgggaa tcctgctctg cgaggctggc cggctaccgc cggcgtaaca gatgagggca
     1921 agcggatggc tgatgaaacc aagccaacca ggaagggcag cccacctatc aaggtgtact
     1981 gccttccaga cgaacgaaga gcgattgagg aaaaggcggc ggcggccggc atgagcctgt
     2041 cggcctacct gctggccgtc ggccagggct acaaaatcac gggcgtcgtg gactatgagc
     2101 acgtccgcga gctggcccgc atcaatggcg acctgggccg cctgggcggc ctgctgaaac
     2161 tctggctcac cgacgacccg cgcacggcgc ggttcggtga tgccacgatc ctcgccctgc
     2221 tggcgaagat cgactctagc tagaaatatt ttatctgatt aataagatga tcttcttgag
     2281 atcgttttgg tctgcgcgta atctcttgct ctgaaaacga aaaaaccgcc ttgcagggcg
     2341 gtttttcgaa ggttctctga gctaccaact ctttgaaccg aggtaactgg cttggaggag
     2401 cgcagtcacc aaaacttgtc ctttcagttt agccttaacc ggcgcatgac ttcaagacta
     2461 actcctctaa atcaattacc agtggctgct gccagtggtg cttttgcatg tctttccggg
     2521 ttggactcaa gacgatagtt accggataag gcgcagcggt cggactgaac ggggggttcg
     2581 tgcatacagt ccagcttgga gcgaactgcc tacccggaac tgagtgtcag gcgtggaatg
     2641 agacaaacgc ggccataaca gcggaatgac accggtaaac cgaaaggcag gaacaggaga
     2701 gcgcacgagg gagccgccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2761 ccaccactga tttgagcgtc agatttcgtg atgcttgtca ggggggcgga gcctatggaa
     2821 aaacggcttt gccgcggccc tctcacttcc ctgttaagta tcttcctggc atcttccagg
     2881 aaatctccgc cccgttcgta agccatttcc gctcgccgca gtcgaacgac cgagcgtagc
     2941 gagtcagtga gcgaggaagc ggaatatatc ctgtatcaca tattctgctg acgcaccggt
     3001 gcagcctttt ttctcctgcc acatgaagca cttcactgac accctcatca gtgccaacat
     3061 agtaagccag tatacactcc gctagcgctg atgtccggcg gtgcttttgc cgttacgcac
     3121 caccccgtca gtagctgaac aggagggaca gttactctag ggcggcggat ttgtcctact
     3181 caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt tcgactgagc
     3241 ctttcgtttt atttgatgcc
//
