LOCUS       sequence-489466-        3217 bp    DNA     circular SYN 16-OCT-2025
DEFINITION  BEVA Golden Gate cloning vector; Narrow host range plasmid with FRT 
            site. L1-p15A-Gm-FRT-ELT4; GG assembly from pNDGG021, pOGG009, 
            pNDGG006, pNDGG009, pOGG014. NmR..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3217)
  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 3217)
  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: pNDGG013
FEATURES             Location/Qualifiers
     source          1..3217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     560..579
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     oriT            830..939
                     /note="incP origin of transfer"
     rep_origin      1322..1867
                     /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     1768..1787
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1984..2001
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2202..2249
                     /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    2334..2355
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2370..2375,2376..2393,2394..2400)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2408..2424
                     /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     2413..2435
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2432..2448
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2432..2448
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2444..2767
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    2461..2517
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2518..2535)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2518..2534)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2527..2549)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2743..2762)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             join(3113..3217,1..690)
                     /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(3167..3186)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
       61 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
      121 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
      181 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
      241 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
      301 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
      361 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
      421 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
      481 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
      541 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
      601 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
      661 ttctatcgcc ttcttgacga gttcttctga actacttttc cgctgcataa ccctgcttcg
      721 gggtcattat agcgattttt tcggtatatc catccttttt cgcacgatat acaggatttt
      781 gccaaagggt tcgtgtagac tttccttggt gtatccaacg gcgtcagccg ggcaggatag
      841 gtgaagtagg cccacccgcg agcgggtgtt ccttcttcac tgtcccttat tcgcacctgg
      901 cggtgctcaa cgggaatcct gctctgcgag gctggccggc taccgccggc gtaacagatg
      961 agggcaagcg gatggctgat gaaaccaagc caaccaggaa gggcagccca cctatcaagg
     1021 tgtactgcct tccagacgaa cgaagagcga ttgaggaaaa ggcggcggcg gccggcatga
     1081 gcctgtcggc ctacctgctg gccgtcggcc agggctacaa aatcacgggc gtcgtggact
     1141 atgagcacgt ccgcgagctg gcccgcatca atggcgacct gggccgcctg ggcggcctgc
     1201 tgaaactctg gctcaccgac gacccgcgca cggcgcggtt cggtgatgcc acgatcctcg
     1261 ccctgctggc gaagatcgac tctagctaga aatattttat ctgattaata agatgatctt
     1321 cttgagatcg ttttggtctg cgcgtaatct cttgctctga aaacgaaaaa accgccttgc
     1381 agggcggttt ttcgaaggtt ctctgagcta ccaactcttt gaaccgaggt aactggcttg
     1441 gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc ttaaccggcg catgacttca
     1501 agactaactc ctctaaatca attaccagtg gctgctgcca gtggtgcttt tgcatgtctt
     1561 tccgggttgg actcaagacg atagttaccg gataaggcgc agcggtcgga ctgaacgggg
     1621 ggttcgtgca tacagtccag cttggagcga actgcctacc cggaactgag tgtcaggcgt
     1681 ggaatgagac aaacgcggcc ataacagcgg aatgacaccg gtaaaccgaa aggcaggaac
     1741 aggagagcgc acgagggagc cgccaggggg aaacgcctgg tatctttata gtcctgtcgg
     1801 gtttcgccac cactgatttg agcgtcagat ttcgtgatgc ttgtcagggg ggcggagcct
     1861 atggaaaaac ggctttgccg cggccctctc acttccctgt taagtatctt cctggcatct
     1921 tccaggaaat ctccgccccg ttcgtaagcc atttccgctc gccgcagtcg aacgaccgag
     1981 cgtagcgagt cagtgagcga ggaagcggaa tatatcctgt atcacatatt ctgctgacgc
     2041 accggtgcag ccttttttct cctgccacat gaagcacttc actgacaccc tcatcagtgc
     2101 caacatagta agccagtata cactccgcta gcgctgatgt ccggcggtgc ttttgccgtt
     2161 acgcaccacc ccgtcagtag ctgaacagga gggacagtta cgaagttcct attccgaagt
     2221 tcctattctc tagaaagtat aggaacttca gggatccgtc gaccagagga gtgagaccgc
     2281 agctggcacg acaggtttgc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     2341 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     2401 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     2461 aagcttgcat gcctgcaggt cgactctaga ggatccccgg gtaccgagct cgaattcact
     2521 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     2581 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     2641 ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac
     2701 gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc
     2761 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     2821 tctgctcccg gcatccgctt acagacaagc tgtgacggtc tcacgctgca atatctggac
     2881 aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc agtgggctta catggcgata
     2941 gctagactgg gcggttttat ggacagcaag cgaaccggaa ttgccagctg gggcgccctc
     3001 tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc caaggatctg
     3061 atggcgcagg ggatcaagat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     3121 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3181 ctgggcacaa cagacaatcg gctgctctga tgccgcc
//
