LOCUS       sequence-351272-        6354 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 6354)
  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 6354)
  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: pFTK061
FEATURES             Location/Qualifiers
     source          1..6354
                     /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"
     CDS             2235..6338
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             2235..6338
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
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 tcaaatggac aagaagtatt ctatcggact ggatatcggg actaatagcg
     2281 tcgggtgggc cgtgatcact gacgagtaca aggtgccctc taagaagttc aaggtgctcg
     2341 ggaacaccga ccggcattcc atcaagaaaa atctgatcgg agctctcctc tttgattcag
     2401 gggaaaccgc tgaagcaacc cgcctcaagc ggactgctag acggcggtac accaggagga
     2461 agaaccggat ttgttacctt caagagatat tctccaacga aatggcaaag gtcgacgaca
     2521 gcttcttcca taggctggaa gaatcattcc tcgtggaaga ggataagaag catgaacggc
     2581 atcccatctt cggtaatatc gtcgacgagg tggcctatca cgagaaatac ccaaccatct
     2641 accatcttcg caaaaagctg gtggactcaa ccgacaaggc agacctccgg cttatctacc
     2701 tggccctggc ccacatgatc aagttcagag gccacttcct gatcgagggc gacctcaatc
     2761 ctgacaatag cgatgtggat aaactgttca tccagctggt gcagacttac aaccagctct
     2821 ttgaagagaa ccccatcaat gcaagcggag tcgatgccaa ggccattctg tcagcccggc
     2881 tgtcaaagag ccgcagactt gagaatctta tcgctcagct gccgggtgaa aagaaaaatg
     2941 gactgttcgg gaacctgatt gctctttcac ttgggctgac tcccaatttc aagtctaatt
     3001 tcgacctggc agaggatgcc aagctgcaac tgtccaagga cacctatgat gacgatctcg
     3061 acaacctcct ggcccagatc ggtgaccaat acgccgacct tttccttgct gctaagaatc
     3121 tttctgacgc catcctgctg tctgacattc tccgcgtgaa cactgaaatc accaaggccc
     3181 ctctttcagc ttcaatgatt aagcggtatg atgagcacca ccaggacctg accctgctta
     3241 aggcactcgt ccggcagcag cttccggaga agtacaagga aatcttcttt gaccagtcaa
     3301 agaatggata cgccggctac atcgacggag gtgcctccca agaggaattt tataagttta
     3361 tcaaacctat ccttgagaag atggacggca ccgaagagct cctcgtgaaa ctgaatcggg
     3421 aggatctgct gcggaagcag cgcactttcg acaatgggag cattccccac cagatccatc
     3481 ttggggagct tcacgccatc cttcggcgcc aagaggactt ctaccccttt cttaaggaca
     3541 acagggagaa gattgagaaa attctcactt tccgcatccc ctactacgtg ggacccctcg
     3601 ccagaggaaa tagccggttt gcttggatga ccagaaagtc agaagaaact atcactccct
     3661 ggaacttcga agaggtggtg gacaagggag ccagcgctca gtcattcatc gaacggatga
     3721 ctaacttcga taagaacctc cccaatgaga aggtcctgcc gaaacattcc ctgctctacg
     3781 agtactttac cgtgtacaac gagctgacca aggtgaaata tgtcaccgaa gggatgagga
     3841 agcccgcatt cctgtcaggc gaacaaaaga aggcaattgt ggaccttctg ttcaagacca
     3901 atagaaaggt gaccgtgaag cagctgaagg aggactattt caagaaaatt gaatgcttcg
     3961 actctgtgga gattagcggg gtcgaagatc ggttcaacgc aagcctgggt acttaccatg
     4021 atctgcttaa gatcatcaag gacaaggatt ttctggacaa tgaggagaac gaggacatcc
     4081 ttgaggacat tgtcctgact ctcactctgt tcgaggaccg ggaaatgatc gaggagaggc
     4141 ttaagaccta cgcccatctg ttcgacgata aagtgatgaa gcaacttaaa cggagaagat
     4201 ataccggatg gggacgcctt agccgcaaac tcatcaacgg aatccgggac aaacagagcg
     4261 gaaagaccat tcttgatttc cttaagagcg acggattcgc taatcgcaac ttcatgcaac
     4321 ttatccatga tgattccctg acctttaagg aggacatcca gaaggcccaa gtgtctggac
     4381 aaggtgactc actgcacgag catatcgcaa atctggctgg ttcacccgct attaagaagg
     4441 gtattctcca gaccgtgaaa gtcgtggacg agctggtcaa ggtgatgggt cgccataaac
     4501 cagagaacat tgtcatcgag atggccaggg aaaaccagac tacccagaag ggacagaaga
     4561 acagcaggga gcggatgaaa agaattgagg aagggattaa ggagctcggg tcacagatcc
     4621 ttaaagagca cccggtggaa aacacccagc ttcagaatga gaagctctat ctgtactacc
     4681 ttcaaaatgg acgcgatatg tatgtggacc aagagcttga tatcaacagg ctctcagact
     4741 acgacgtgga ccatatcgtc cctcagagct tcctcaaaga cgactcaatt gacaataagg
     4801 tgctgactcg ctcagacaag aaccggggaa agtcagataa cgtgccctca gaggaagtcg
     4861 tgaaaaagat gaagaactat tggcgccagc ttctgaacgc aaagctgatc actcagcgga
     4921 agttcgacaa tctcactaag gctgagaggg gcggactgag cgaactggac aaagcaggat
     4981 tcattaaacg gcaacttgtg gagactcggc agattactaa acatgtcgcc caaatccttg
     5041 actcacgcat gaataccaag tacgacgaaa acgacaaact tatccgcgag gtgaaggtga
     5101 ttaccctgaa gtccaagctg gtcagcgatt tcagaaagga ctttcaattc tacaaagtgc
     5161 gggagatcaa taactatcat catgctcatg acgcatatct gaatgccgtg gtgggaaccg
     5221 ccctgatcaa gaagtaccca aagctggaaa gcgagttcgt gtacggagac tacaaggtct
     5281 acgacgtgcg caagatgatt gccaaatctg agcaggagat cggaaaggcc accgcaaagt
     5341 acttcttcta cagcaacatc atgaatttct tcaagaccga aatcaccctt gcaaacggtg
     5401 agatccggaa gaggccgctc atcgagacta atggggagac tggcgaaatc gtgtgggaca
     5461 agggcagaga tttcgctacc gtgcgcaaag tgctttctat gcctcaagtg aacatcgtga
     5521 agaaaaccga ggtgcaaacc ggaggctttt ctaaggaatc aatcctcccc aagcgcaact
     5581 ccgacaagct cattgcaagg aagaaggatt gggaccctaa gaagtacggc ggattcgatt
     5641 caccaactgt ggcttattct gtcctggtcg tggctaaggt ggaaaaagga aagtctaaga
     5701 agctcaagag cgtgaaggaa ctgctgggta tcaccattat ggagcgcagc tccttcgaga
     5761 agaacccaat tgactttctc gaagccaaag gttacaagga agtcaagaag gaccttatca
     5821 tcaagctccc aaagtatagc ctgttcgaac tggagaatgg gcggaagcgg atgctcgcct
     5881 ccgctggcga acttcagaag ggtaatgagc tggctctccc ctccaagtac gtgaatttcc
     5941 tctaccttgc aagccattac gagaagctga aggggagccc cgaggacaac gagcaaaagc
     6001 aactgtttgt ggagcagcat aagcattatc tggacgagat cattgagcag atttccgagt
     6061 tttctaaacg cgtcattctc gctgatgcca acctcgataa agtccttagc gcatacaata
     6121 agcacagaga caaaccaatt cgggagcagg ctgagaatat catccacctg ttcaccctca
     6181 ccaatcttgg tgcccctgcc gcattcaagt acttcgacac caccatcgac cggaaacgct
     6241 atacctccac caaagaagtg ctggacgcca ccctcatcca ccagagcatc accggacttt
     6301 acgaaactcg gattgacctc tcacagctcg gaggggatag gttgagacca cgaa
//
