LOCUS       sequence-462968-        3425 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BEVA Golden Gate cloning vector; BEVA2.0 Narrow host range plasmid 
            with I-SceI counterselectable site. L1-P15A-Nm-ISceI; GG assembly 
            from pOGG004, pNDGG001, pNDGG006, pGQ0023, pOGG014. KmR/NmR..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3425)
  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 3425)
  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: pQGG002
FEATURES             Location/Qualifiers
     source          1..3425
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    83..104
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(119..124,125..142,143..149)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    157..173
                     /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     162..184
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     181..197
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     181..197
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             193..516
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    210..266
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(267..284)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(267..283)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(276..298)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(492..511)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      621..715
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             965..1759
                     /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(1019..1038)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1629..1648
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     oriT            1899..2008
                     /note="incP origin of transfer"
     rep_origin      2391..2936
                     /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     2837..2856
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3053..3070
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3334..3420)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tgccgaattc ggatccggag tgagaccgca gctggcacga caggtttgcc gactggaaag
       61 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
      121 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
      181 caggaaacag ctatgaccat gattacgcca agcttgcatg cctgcaggtc gactctagag
      241 gatccccggg taccgagctc gaattcactg gccgtcgttt tacaacgtcg tgactgggaa
      301 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
      361 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
      421 tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
      481 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
      541 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
      601 gtgacggtct cacgctcttg gactcctgtt gatagatcca gtaatgacct cagaactcca
      661 tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccagg
      721 caatatctgg acaagggaaa acgcaagcgc aaagagaaag caggtagctt gcagtgggct
      781 tacatggcga tagctagact gggcggtttt atggacagca agcgaaccgg aattgccagc
      841 tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc
      901 gccaaggatc tgatggcgca ggggatcaag atctgatcaa gagacaggat gaggatcgtt
      961 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     1021 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     1081 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     1141 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     1201 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     1261 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     1321 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     1381 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     1441 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     1501 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     1561 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     1621 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     1681 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     1741 tcttgacgag ttcttctgaa ctacttttcc gctgcataac cctgcttcgg ggtcattata
     1801 gcgatttttt cggtatatcc atcctttttc gcacgatata caggattttg ccaaagggtt
     1861 cgtgtagact ttccttggtg tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc
     1921 ccacccgcga gcgggtgttc cttcttcact gtcccttatt cgcacctggc ggtgctcaac
     1981 gggaatcctg ctctgcgagg ctggccggct accgccggcg taacagatga gggcaagcgg
     2041 atggctgatg aaaccaagcc aaccaggaag ggcagcccac ctatcaaggt gtactgcctt
     2101 ccagacgaac gaagagcgat tgaggaaaag gcggcggcgg ccggcatgag cctgtcggcc
     2161 tacctgctgg ccgtcggcca gggctacaaa atcacgggcg tcgtggacta tgagcacgtc
     2221 cgcgagctgg cccgcatcaa tggcgacctg ggccgcctgg gcggcctgct gaaactctgg
     2281 ctcaccgacg acccgcgcac ggcgcggttc ggtgatgcca cgatcctcgc cctgctggcg
     2341 aagatcgact ctagctagaa atattttatc tgattaataa gatgatcttc ttgagatcgt
     2401 tttggtctgc gcgtaatctc ttgctctgaa aacgaaaaaa ccgccttgca gggcggtttt
     2461 tcgaaggttc tctgagctac caactctttg aaccgaggta actggcttgg aggagcgcag
     2521 tcaccaaaac ttgtcctttc agtttagcct taaccggcgc atgacttcaa gactaactcc
     2581 tctaaatcaa ttaccagtgg ctgctgccag tggtgctttt gcatgtcttt ccgggttgga
     2641 ctcaagacga tagttaccgg ataaggcgca gcggtcggac tgaacggggg gttcgtgcat
     2701 acagtccagc ttggagcgaa ctgcctaccc ggaactgagt gtcaggcgtg gaatgagaca
     2761 aacgcggcca taacagcgga atgacaccgg taaaccgaaa ggcaggaaca ggagagcgca
     2821 cgagggagcc gccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     2881 actgatttga gcgtcagatt tcgtgatgct tgtcaggggg gcggagccta tggaaaaacg
     2941 gctttgccgc ggccctctca cttccctgtt aagtatcttc ctggcatctt ccaggaaatc
     3001 tccgccccgt tcgtaagcca tttccgctcg ccgcagtcga acgaccgagc gtagcgagtc
     3061 agtgagcgag gaagcggaat atatcctgta tcacatattc tgctgacgca ccggtgcagc
     3121 cttttttctc ctgccacatg aagcacttca ctgacaccct catcagtgcc aacatagtaa
     3181 gccagtatac actccgctag cgctgatgtc cggcggtgct tttgccgtta cgcaccaccc
     3241 cgtcagtagc tgaacaggag ggacagttac tgaccattcc aaggtaggga taacagggta
     3301 atttggaacc gtagtccaga tctagggcgg cggatttgtc ctactcagga gagcgttcac
     3361 cgacaaacaa cagataaaac gaaaggccca gtctttcgac tgagcctttc gttttatttg
     3421 atgcc
//
