LOCUS       sequence-254496-       10151 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AMA1 plasmid with ble selection marker, and USER cassette 
            (PacI/Nt.BbvCI).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10151)
  AUTHORS   Vanegas KG, Jarczynska ZD, Strucko T, Mortensen UH
  TITLE     Cpf1 enables fast and efficient genome editing in Aspergilli.
  JOURNAL   Fungal Biol Biotechnol. 2019 May 1;6:6. doi: 
            10.1186/s40694-019-0069-6. eCollection 2019.
  PUBMED    31061713
REFERENCE   2  (bases 1 to 10151)
  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: "Fungal Biol
            Biotechnol."; date: "2019-05-1"; volume: "6:6. doi"; pages: " 
            10.1186/s40694-019-0069-6. eCollection 2019"
COMMENT     Sequence Label: pAC575
FEATURES             Location/Qualifiers
     source          1..10151
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    189..222
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     rep_origin      535..5783
                     /label=AMA1
                     /note="replication origin from Aspergillus nidulans 
                     (Aleksenko and Clutterbuck, 1996)"
     CDS             6393..6767
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     protein_bind    complement(7596..7629)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(8099..8115)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8214..8231)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8385..8973)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8465..8484)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9095..9886,9887..9955))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     9718..9737
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9956..10047)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     10089..10108
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     10114..10133
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg
       61 tagtgtgggg actccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg
      121 ctcagtcgaa agactgggcc tttcgcccgg gctaattatg gggtgtcgcc cttattcgac
      181 tctatagtga agttcctatt ctctagaaag tataggaact tctgaagtgg ggatttaaat
      241 gcggccgcgc tgagggttta atcgacgaag cagctgacgg ccagtgccaa gcttaacgcg
      301 taccgggccc agtatatgtt ccgcagatga ctggagctct gccatacgtg ccctctcaag
      361 caccatttgt tccatctaca gagactagtc accaactagt ctatcaagac tcacagggta
      421 cattgctgag accaactgac cagaggcagg gtagcggatt gacggctcca tctccttcac
      481 ttacaaggtc tattgaaagc cctttagcat caccaagcgg agaatagatt gttaagctta
      541 ttttttgtat actgttttgt gatagcacga agtttttcca cggtatcttg ttaaaaatat
      601 atatttgtgg cgggcttacc tacatcaaat taataagaga ctaattataa actaaacaca
      661 caagcaagct actttagggt aaaagtttat aaatgctttt gacgtataaa cgttgcttgt
      721 atttattatt acaattaaag gtggatagaa aacctagaga ctagttagaa actaatctca
      781 ggtttgcgtt aaactaaatc agagcccgag aggttaacag aacctagaag gggactagat
      841 atccgggtag ggaaacaaaa aaaaaaaaca agacakccac atattaggga gactagttag
      901 aagctagttc caggactagg aaaataaaag acaatgatac cacagtctag ttgacaacta
      961 gatagattct agattgaggc caaagtctct gagatccagg ttagttgcaa ctaatactag
     1021 ttagtatcta gtctcctata actctgaagc tagaataact tactactatt atcctcacca
     1081 ctgttcagct gcgcaaacgg agtgattgca aggtgttcag agactagtta ttgactagtc
     1141 agtgactagc aataactaac aaggtattaa cctaccatgt ctgccatcac cctgcacttc
     1201 ctcgggctca gcagcctttt cctcctcatt ttcatgctca ttttccttgt ttaagactgt
     1261 gactagtcaa agactagtcc agaaccacaa aggagaaatg tcttaccact ttcttcattg
     1321 cttgtctctt ttgcattatc catgtctgca actagttaga gtctagttag tgactagtcc
     1381 gacgaggact tgcttgtctc cggattgttg gaggaactct ccagggcctc aagatccaca
     1441 acagagcctt ctagaagact ggtcaataac tagttggtct ttgtctgagt ctgacttacg
     1501 aggttgcata ctcgctccct ttgcctcgtc aatcgatgag aaaaagcgcc aaaactcgca
     1561 atatggcttt gaaccacacg gtgctgagac tagttagaat ctagtcccaa actagcttgg
     1621 atagcttacc tttgyccttt gcgttgcgac aggtcttgca gggtatggtt cctttctcac
     1681 cagctgattt agctgccttg ctaccctcac ggcggatctg cataaagagt ggctagaggt
     1741 tataaattag cactgatcct aggtacgggg ctgaatgtaa cttgccyttc ctttctcatc
     1801 gcgcggcaag acaggcttgc tcaaattcct accagtcaca ggggtatgca cggcgtacgg
     1861 accacttgaa ctagtcacag attagttagc aactagtctg cattgaatgg ctgtacttac
     1921 gggccctcgc cattgtcctg atcatttcca gcttcaccct cgttgctgca aagtagttag
     1981 tgactagtca aggactagtt gaaatgggag aagaaactca cgaattctcg acacccttag
     2041 tattgtggtc cttggacttg gtgctgctat atattagcta atacactagt tagactcaca
     2101 gaaacttacg cagctcgctt gcgcttcttg gtaggagtcg gggttgggag aacagtgcct
     2161 tcaaacaagc cttcatacca tgctacttga ctagtcaggg actagtcacc aagtaatcta
     2221 gataggactt gcctttggcc tccatcagtt ccttcatagt gggaggtcca ttgtgcaatg
     2281 taaactccat gccgtgggag ttcttgtcct tcaagtgctt gaccaatatg tttctgttgg
     2341 cagagggaac ctgtcaacta gttaataact agtcagaaac tagtatagca gtagactcac
     2401 tgtacgcttg aggcatccct tcactcggca gtagacttca tatggatgga tatcaggcac
     2461 gccattgtcg tcctgtggac tagtcagtaa ctaggcttaa agctagtcgg gtcggcttac
     2521 tatcttgaaa tccggcagcg taagctcccc gtccttaact gcctcgagat agtgacagta
     2581 ctctggggac tttcggagat cgttatcgcg aatgctcggc atactaatcg ttgactagtc
     2641 ttggactagt cccgagcaaa aaggattgga ggagraggag gaaggtgaga gtgagacaaa
     2701 gagcgaaata agagcttcaa aggctatctc taagcagtat gaaggttaag tatctagttc
     2761 ttgactagat ttaaaagaga tttcgactag ttatgtacct ggagtttgga tataggaatg
     2821 tgttgtggta acgaaatgta agggggagga aagaaaaagt cggtcaatag gtaactctaa
     2881 gtcggccatt cctttttggg agtcgctaac cataaacggc atggtcgact tagagttagc
     2941 tcagggaatt tagggagtta tctgcgacca ccgaggaacg gcggaatgcc aaagaatccc
     3001 gatggagctc tagctggcgg ttgacaaccc caccttttgg cgtttctgcg gcgttgcagg
     3061 cgggactgga tacttcgtag aaccagaaag gcaaggcaga acgcgctcag caagagtgtt
     3121 ggaagtgata gcatgatgtg ccttgttaac taggtcaaaa tctgcagtat gcttgatgtt
     3181 atccaaagtg tgagagagga aggtccaaac atacacgatt gggagagggc ctaggtataa
     3241 gagtttttga gtagaacgca tgtgagccca gccatctcga ggagattaaa cacgggccgg
     3301 catttgatgg ctatgttagt accccaatgg aaacggtgag agtccagtgg tcgcagataa
     3361 ctccctaaat tccctgagct aactctaagt cgaccatgcc gtttatggtt agcgcctccc
     3421 aaaaaggaat ggccgactta gagttacctc ttgaccgact ttttctttcc tcccccttac
     3481 atttcgttac cacaacacat tcctatatcc aaactccagg tacataacta gtcgaaatct
     3541 cttttaaatc tagtcaagaa ctagatactt aaccttcata ctgcttagag atagcctttg
     3601 aagctcttat ttcgctcttt gtctcactct caccttcctc ctcctcctcc aatccttttt
     3661 gctcgggact agtccaagac tagtcaacga ttagtatgcc gagcattcgc gataacgatc
     3721 tccgaaagtc cccagagtac tgtcactatc tcgaggcagt taaggacggg gagcttacgc
     3781 tgccggattt caagatagta agccgacccg actagcttta agcctagtta ctgactagtc
     3841 cacaggacga caatggcgtg cctgatatcc atccatatga agtctactgc cgagtgaagg
     3901 gatgcctcaa gcgtacagtg agtctactgc tatactagtt tctgactagt tattaactag
     3961 ttgacaggtt ccctctgcca acagaaacat attggtcaag cacttgaagg acaagaactc
     4021 ccacggcatg gagtttacat tgcacaatgg acctcccact atgaaggaac tgatggaggc
     4081 caaaggcaag tcctatctag attacttggt gactagtccc tgactagtca agtagcatgg
     4141 tatgaaggct tgtttgaagg cactgttctc ccaaccccga ctcctaccaa gaagcgcaag
     4201 cgagctgcgt aagtttctgt gagtctaact agtgtattag ctaatatata gcagcaccaa
     4261 gtccaaggac cacaatacta agggtgtcga gaattcgtga gtttcttctc ccatttcaac
     4321 tagtccttga ctagtcacta actactttgc agcaacgagg gtgaagctgg aaatgatcag
     4381 gacaatggcg agggcccgta agtacagcca ttcaatgcag actagttgct aactaatctg
     4441 tgactagttc aagtggtccg tacgccgtgc atacccctgt gactggtagg aatttgagca
     4501 agcctgtctt gccgcgcgat gagaaaggaa rggcaagtta cattcagccc cgtacctagg
     4561 atcagtgcta atttataacc tctagccact ctttatgcag atccgccgtg agggtagcaa
     4621 ggcagctaaa tcagctggtg agaaaggaac cataccctgc aagacctgtc gcaacgcaaa
     4681 ggrcaaaggt aagctatcca agctagtttg ggactagatt ctaactagtc tcagcaccgt
     4741 gtggttcaaa gccatattgc gagttttggc gctttttctc atcgattgac gaggcaaagg
     4801 gagcgagtat gcaacctcgt aagtcagact cagacaaaga ccaactagtt attgaccagt
     4861 cttctagaag gctctgttgt ggatcttgag gccctggaga gttcctccaa caatccggag
     4921 acaagcaagt cctcgtcgga ctagtcacta actagactct aactagttgc agacatggat
     4981 aatgcaaaag agacaagcaa tgaagaaagt ggtaagacat ttctcctttg tggttctgga
     5041 ctagtctttg actagtcaca gtcttaaaca aggaaaatga gcatgaaaat gaggaggaaa
     5101 aggctgctga gcccgaggaa gtgcagggtg atggcagaca tggtaggtta ataccttgtt
     5161 agttattgct agtcactgac tagtcaataa ctagtctctg aacaccttgc aatcactccg
     5221 tttgcgcagc tgaacagtgg tgaggataat agtagtaagt tattctagct tcagagttat
     5281 aggagactag atactaacta gtattagttg caactaacct ggatctcaga gactttggcc
     5341 tcaatctaga atctatctag ttgtcaacta gactgtggta tcattgtctt ttattttcct
     5401 agtcctggaa ctagcttcta actagtctcc ctaatatgtg gmtgtcttgt tttttttttt
     5461 tgtttcccta cccggatatc tagtcccctt ctaggttctg ttaacctctc gggctctgat
     5521 ttagtttaac gcaaacctga gattagtttc taactagtct ctaggttttc tatccacctt
     5581 taattgtaat aataaataca agcaacgttt atacgtcaaa agcatttata aacttttacc
     5641 ctaaagtagc ttgcttgtgt atttagttta taattagtct cttattaatt tgatgtaggt
     5701 aagcccgcca caaatatata tttttaacaa gataccgtgg aaaaacttcg tgctatcaca
     5761 aaacagtata caaaaaataa gcttaacaat ctattctccg cttggtgatg ctaaagggct
     5821 ttcaatagac cttgtaagtg aaggagatgg agccgtcaat ccgctaccct gcctctggtc
     5881 agttggtctc agcaatgtac cctgtgagtc ttgatagact agttggtgac tagtctctgt
     5941 agatggaaca aatggtgctt gagagggcac gtatggcaga gctccagtca tctgcggaac
     6001 atatactggg cccgggaaga tctcatggtc atagctgttt ccgctgaggg tttaattaag
     6061 acctcagcgc tagtggaggt caacacatca atgcctattt tggtttagtc gtccaggcgg
     6121 tgagcacaaa atttgtgtcg tttgacaaga tggttcattt aggcaactgg tcagatcagc
     6181 cccacttgta gcagtagcgg cggcgctcga agtgtgactc ttattagcag acaggaacga
     6241 ggacattatt atcatctgct gcttggtgca cgataacttg gtgcgtttgt caagcaaggt
     6301 aagtggacga cccggtcata ccttcttaag ttcgcccttc ctccctttat ttcagattca
     6361 atctgactta cctattctac ccaagcatcc aaatggccaa gttgaccagt gccgttccgg
     6421 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     6481 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
     6541 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     6601 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     6661 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     6721 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgaccg acgccgacca
     6781 acaccgccgg tccgacgcgg cccgacgggt ccgaggagct tgagatccac ttaacgttac
     6841 tgaaatcatc aaacagcttg acgaatctgg atataagatc gttggtgtcg atgtcagctc
     6901 cggagttgag acaaatggtg ttcaggatct cgataagata cgttcatttg tccaagcagc
     6961 aaagagtgcc ttctagtgat ttaatagctc catgtcaaca agaataaaac gcgtttcggg
     7021 tttacctctt ccagatacag ctcatctgca atgcattaat gcattggacc tcgcaaccct
     7081 agtacgccct tcaggctccg gcgaagcaga agaatagctt agcagagtct attttcattt
     7141 tcgggagacg agatcaagca gatcaacggt cgtcaagaga cctacgagac tgaggaatcc
     7201 gctcttggct ccacgcgact atatatttgt ctctaattgt actttgacat gctcctcttc
     7261 tttactctga tagcttgact atgaaaattc cgtcaccagc ccctgggttc gcaaagataa
     7321 ttgcactgtt tcttccttga actctcaagc ctacaggaca cacattcatc gtaggtataa
     7381 acctcgaaaa tcattcctac taagatgggt atacaatagt aaccatgcat ggttgcctag
     7441 tgaatgctcc gtaacaccca atacgccggc cgaaactttt ttacaactct cctatgagtc
     7501 gtttacccag aatgcacagg tacacttgtt tagaggtaaa tccccactac cgcattaaga
     7561 cctcagcgcg gccgcaaatt taaataaaat gaagtgaagt tcctatactt tctagagaat
     7621 aggaacttct atagtgagtc gaataagggc gacacaaaat ttattctaaa tgcataataa
     7681 atactgataa catcttatag tttgtattat attttgtatt atcgttgaca tgtataattt
     7741 tgatatcaaa aactgatttt ccctttatta ttttcgagat ttattttctt aattctcttt
     7801 aacaaactag aaatattgta tatacaaaaa atcataaata atagatgaat agtttaatta
     7861 taggtgttca tcaatcgaaa aagcaacgta tcttatttaa agtgcgttgc ttttttctca
     7921 tttataaggt taaataattc tcatatatca agcaaagtga caggcgccct taaatattct
     7981 gacaaatgct ctttccctaa actcccccca taaaaaaacc cgccgaagcg ggtttttacg
     8041 ttatttgcgg attaacgatt actcgttatc agaaccgccc agggggcccg agcttaagac
     8101 tggccgtcgt tttacaacac agaaagagtt tgtagaaacg caaaaaggcc atccgtcagg
     8161 ggccttctgc ttagtttgat gcctggcagt tccctactct cgccttccgc ttcctcgctc
     8221 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     8281 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     8341 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     8401 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     8461 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     8521 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     8581 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     8641 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     8701 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     8761 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt gggctaacta cggctacact
     8821 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     8881 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     8941 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     9001 tctgacgctc agtggaacga cgcgcgcgta actcacgtta agggattttg gtcatgagct
     9061 tgcgccgtcc cgtcaagtca gcgtaatgct ctgcttacca atgcttaatc agtgaggcac
     9121 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     9181 taactacgat acgggagggc ttaccatctg gccccagcgc tgcgatgata ccgcgagaac
     9241 cacgctcacc ggctccggat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     9301 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     9361 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccatcgct acaggcatcg
     9421 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     9481 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     9541 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagcg ctacataatt
     9601 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     9661 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     9721 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     9781 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     9841 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     9901 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatattct
     9961 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
    10021 ttgaatgtat ttagaaaaat aaacaaatag gggtcagtgt tacaaccaat taaccaattc
    10081 tgaacattat cgcgagccca tttatacctg aatatggctc ataacacccc ttgtttgcct
    10141 ggcggcagta g
//
