LOCUS       sequence-351696-        3733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Part Plasmid Entry Vector (LVL0) from the Fungal Modular Cloning 
            ToolKit..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3733)
  AUTHORS   Mozsik L, Pohl C, Meyer V, Bovenberg RAL, Nygard Y, Driessen AJM
  TITLE     Modular Synthetic Biology Toolkit for Filamentous Fungi.
  JOURNAL   ACS Synth Biol. 2021 Nov 2. doi: 10.1021/acssynbio.1c00260.
  PUBMED    34726388
REFERENCE   2  (bases 1 to 3733)
  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: "ACS Synth 
            Biol. 2021 Nov 2. doi: 10.1021/acssynbio.1c00260."
COMMENT     Sequence Label: pFTK086
FEATURES             Location/Qualifiers
     source          1..3733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(24..42)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        110..214
                     /gene="bla"
                     /label=AmpR promoter
     CDS             436..1227
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1398..1986
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1887..1906
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2140..2157
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2775..2796
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2811..2816,2817..2834,2835..2841)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2849..2865
                     /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     2854..2876
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2873..2889
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2873..2889
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2885..3208
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    2902..2958
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2959..2976)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2959..2975)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2968..2990)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3184..3203)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_RNA        3301..3376
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 gtggctcttc agtggacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
       61 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
      121 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
      181 ataaatgctt caataatatt gaaaaaggaa gagtatgcgc tcacgcaact ggtccagaac
      241 cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg
      301 tttttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg
      361 ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa
      421 acaaagttaa acatcatgag ggaagcggtg atcgccgaag tatcgactca actatcagag
      481 gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc
      541 tccgcagtgg atggcggcct gaagccacac agcgatattg atttgctggt tacggtgacc
      601 gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct
      661 tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac
      721 atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat
      781 gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg
      841 acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat
      901 ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg
      961 ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac
     1021 agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc
     1081 ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa
     1141 gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccatta cgtaaaaggc
     1201 gagatcacca aggtagtcgg caaataactg tcagaccaag tttactcata tatactttag
     1261 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     1321 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1381 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1441 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1501 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     1561 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1621 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1681 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     1741 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     1801 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     1861 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     1921 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     1981 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2041 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2101 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2161 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatca
     2221 ctctgtggtc tcggagttcc atcttagcga tcgggatgta accctgcgac ccccggcatc
     2281 acccaccatt tgtcccctcg tgaatgcttc gttctgtttg ctgtcggatg tagataggct
     2341 ggggcagatg actacctagt acttggtggt atcccacgtc agtcgccgga acgagaacgg
     2401 tgccactggg acaagaggct ctcagacaag taacacagta gcaaatggtt tgactaatga
     2461 gataatgaac caactctgga cgataagtgt ttgcacgaat gataacgctg ggatggtcgt
     2521 ggattccgac tgccccgctg gaagtgctcg gagttgcccc cactcggcac ggtacttccg
     2581 gtgaccaaaa cggtaccttt ggctcatata acaaattgct accatcattt cccatcaaat
     2641 aggcctgctg gcccaatggt aaggcgcttg actacggatc aagagattgc aggttcgagt
     2701 cctgcgtagg tcatgagacg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     2761 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     2821 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     2881 agctatgacc atgattacgc caagcttgca tgcctgcagg tcgactctag aggatccccg
     2941 ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     3001 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     3061 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct
     3121 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     3181 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     3241 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgcgtctca
     3301 gtttcagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
     3361 ggcaccgagt cggtgcgttt ctttttcttt tttagtagat gccgaccggg atccacttaa
     3421 cgttactgaa atcatcaaac agcttgacga atctggatat aagatcgttg gtgtcgatgt
     3481 cagctccgga gttgagacaa atggtgttca ggatctcgat aagatacgtt catttgtcca
     3541 agcagcaaag agtgccttct agtgatttaa tagctccatg tcaacaagaa taaaacgcgt
     3601 ttcgggttta cctcttccag atacagctca tctgcaatgc attaatgcat tggacctcgc
     3661 aaccctagta cgcccttcag gctccggcga agcagaagaa tagcttagca gagctatcgc
     3721 ttgagaccac gaa
//
