LOCUS       sequence-254277-       10240 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AMA1 plasmid with pyrG selection marker, and USER cassette 
            (PacI/Nt.BbvCI).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10240)
  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 10240)
  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: pAC572
FEATURES             Location/Qualifiers
     source          1..10240
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     622..641
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(860..951)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     993..1012
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1018..1037
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    1244..1277
                     /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      1590..6840
                     /label=AMA1
                     /note="replication origin from Aspergillus nidulans 
                     (Aleksenko and Clutterbuck, 1996)"
     protein_bind    complement(8740..8773)
                     /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(9243..9259)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9358..9375)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9529..10117)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9609..9628)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10239..10240,1..790,791..859))
                     /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"
ORIGIN
        1 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
       61 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
      121 gcgctgcgat gataccgcga gaaccacgct caccggctcc ggatttatca gcaataaacc
      181 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
      241 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
      301 ttgttgccat cgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
      361 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
      421 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
      481 tggttatggc agcgctacat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
      541 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
      601 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
      661 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
      721 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
      781 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
      841 ggaaatgttg aatactcata ttcttccttt ttcaatatta ttgaagcatt tatcagggtt
      901 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggtca
      961 gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata cctgaatatg
     1021 gctcataaca ccccttgttt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg
     1081 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggactcc ccatgcgaga
     1141 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     1201 cccgggctaa ttatggggtg tcgcccttat tcgactctat agtgaagttc ctattctcta
     1261 gaaagtatag gaacttctga agtggggatt taaatgcggc cgcgctgagg gtttaatcga
     1321 cgaagcagct gacggccagt gccaagctta acgcgtaccg ggcccagtat atgttccgca
     1381 gatgactgga gctctgccat acgtgccctc tcaagcacca tttgttccat ctacagagac
     1441 tagtcaccaa ctagtctatc aagactcaca gggtacattg ctgagaccaa ctgaccagag
     1501 gcagggtagc ggattgacgg ctccatctcc ttcacttaca aggtctattg aaagcccttt
     1561 agcatcacca agcggagaat agattgttaa gcttattttt tgtatactgt tttgtgatag
     1621 cacgaagttt ttccacggta tcttgttaaa aatatatatt tgtggcgggc ttacctacat
     1681 caaattaata agagactaat tataaactaa acacacaagc aagctacttt agggtaaaag
     1741 tttataaatg cttttgacgt ataaacgttg cttgtattta ttattacaat taaaggtgga
     1801 tagaaaacct agagactagt tagaaactaa tctcaggttt gcgttaaact aaatcagagc
     1861 ccgagaggtt aacagaacct agaaggggac tagatatccg ggtagggaaa caaaaaaaaa
     1921 aaaacaagac agccacatat tagggagact agttagaagc tagttccagg actaggaaaa
     1981 taaaagacaa tgataccaca gtctagttga caactagata gattctagat tgaggccaaa
     2041 gtctctgaga tccaggttag ttgcaactaa tactagttag tatctagtct cctataactc
     2101 tgaagctaga ataacttact actattatcc tcaccactgt tcagctgcgc aaacggagtg
     2161 attgcaaggt gttcagagac tagttattga ctagtcagtg actagcaata actaacaagg
     2221 tattaaccta ccatgtctgc catcaccctg cacttcctcg ggctcagcag ccttttcctc
     2281 ctcattttca tgctcatttt ccttgtttaa gactgtgact agtcaaagac tagtccagaa
     2341 ccacaaagga gaaatgtctt accactttct tcattgcttg tctcttttgc attatccatg
     2401 tctgcaacta gttagagtct agttagtgac tagtccgacg aggacttgct tgtctccgga
     2461 ttgttggagg aactctccag ggcctcaaga tccacaacag agccttctag aagactggtc
     2521 aataactagt tggtctttgt ctgagtctga cttacgaggt tgcatactcg ctccctttgc
     2581 ctcgtcaatc gatgagaaaa agcgccaaaa ctcgcaatat ggctttgaac cacacggtgc
     2641 tgagactagt tagaatctag tcccaaacta gcttggatag cttacctttg ccctttgcgt
     2701 tgcgacaggt cttgcagggt atggttcctt tctcaccagc tgatttagct gccttgctac
     2761 mctcacggcg gatctgcata aagagtggct agaggttata aattagcact gatcctaggt
     2821 acggggctga atgtaacttg ccyttccttt ctcatcgcgc ggcaagacag gcttgctcaa
     2881 attcctacca gtcacagggg tatgcacggc gtacggacca cttgaactag tcacagatta
     2941 gttagcaact agtctgcatt gaatggctgt acttacgggc cctcgccatt gtcctgatca
     3001 tttccagctt caccctcgtt gctgcaaagt agttagtgac tagtcaagga ctagttgaaa
     3061 tgggagaaga aactcacgaa ttctcgacac ccttagtatt gtggtccttg gacttggtgc
     3121 tgctatatat tagctaatac actagttaga ctcacagaaa cttacgcagc tcgcttgcgc
     3181 ttcttggtag gagtcggggt tgggagaaca gtgccttcaa acaagccttc ataccatgct
     3241 acttgactag tcagggacta gtcaccaagt aatctagata ggacttkcct ttggcctcca
     3301 tcagttcctt catagtggga ggtccattgt gcaatgtaaa ctccatgccg tgggagttct
     3361 tgtccttcaa gtgcttgacc aatatgtttc tgttggcaga gggaacctgt caactagtta
     3421 ataactagtc agaaactagt atagcagtag actcactgta cgcttgaggc atcccttcac
     3481 tcggcagtag acttcatatk gatggatatc aggcacgcca ttgtcgtcct gtggactagt
     3541 cagtaactag gcttaaagct agtcgggtcg gcttactatc ttgaaatccg gcagcgtaag
     3601 ctccccgtcc ttaactgcct cgagatagtg acagtactct ggggactttc ggagatcgtt
     3661 atcgcgaatg ctcggcatac taatcgttga ctagtcttgg actagtcccg agcaaaaagg
     3721 attggaggag gaggaggaag gtgagagtga gacaaagagc gaaataagag cttcaaaggc
     3781 tatctctaag cagtatgaag gttaagtatc tagttcttga ctagatttaa aagagatttc
     3841 gactagttat gtacctggag tttggatata ggaatgtgtt gtggtaacga aatgtaaggg
     3901 ggaggaaaga aaaagtcggt caagaggtaa ctctaagtcg gccattcctt tttgggaggc
     3961 gctaaccata aacggcatgg tcgacttaga gttagctcag ggaatttagg gagttatctg
     4021 cgaccaccga ggaacggctg aatgccaaag aatcccgatg gagctctagc tggcggttga
     4081 caaccccacc ttttggcgtt tctgcggcgt tgcaggcggg actggatact tcgtagaacc
     4141 agaaaggcaa ggcagaacgc gctcagcaag agtgttggaa gtgatagcat gatgtgcctt
     4201 gttaactagg tcaaaatctg cagtatgctt gatgttatcc aaagtgtgag agaggaaggt
     4261 ccaaacatac acgattggga gagggcctag gtataagagt ttttgagtag aacgcatgtg
     4321 agcccagcca tctcgaggag attaaacacg ggccggcatt tgatggctat gttagtaccc
     4381 caatggaaac ggtgagagtc cagtggtcgc agataactcc ctaaattccc tgagctaact
     4441 ctaagtcgac catgccgttt atggttagcg cctcccaaaa aggaatggcc gacttagagt
     4501 tacctcttga ccgacttttt ctttcctccc ccttacattt cgttaccaca acacattcct
     4561 atatccaaac tccaggtaca taactagtcg aaatctcttt taaatctagt caagaactag
     4621 atacttaacc ttcatactgc ttagagatag cctttgaagc tcttatttcg ctctttgtct
     4681 cactctcacc ttcctcctcc tcctccaatc ctttttgctc gggactagtc caagactagt
     4741 caacgattag tatgccgagc attcgcgata acgatctccg aaagtcccca gagtactgtc
     4801 actatctcga ggcagttaag gacggggagc ttacgctgcc ggatttcaag atagtaagcc
     4861 gacccgacta gctttaagcc tagttactga ctagtccaca ggacgacaat ggcgtgcctg
     4921 atatccatcc atatgaagtc tactgccgag tgaagggatg cctcaagcgt acagtgagtc
     4981 tactgctata ctagtttctg actagttatt aactagttga caggttccct ctgccaacag
     5041 aaacatattg gtcaagcact tgaaggacaa gaactcccac ggcatggagt ttacattgca
     5101 caatggacct cccactatga aggaactgat ggaggccaaa ggmaagtcct atctagatta
     5161 cttggtgact agtccctgac tagtcaagta gcatggtatg aaggcttgtt tgaaggcact
     5221 gttctcccaa ccccgactcc taccaagaag cgcaagcgag ctgcgtaagt ttctgtgagt
     5281 ctaactagtg tattagctaa tatatagcag caccaagtcc aaggaccaca atactaaggg
     5341 tgtcgagaat tcgtgagttt cttctcccat ttcaactagt ccttgactag tcactaacta
     5401 ctttgcagca acgagggtga agctggaaat gatcaggaca atggcgaggg cccgtaagta
     5461 cagccattca atgcagacta gttgctaact aatctgtgac tagttcaagt ggtccgtacg
     5521 ccgtgcatac ccctgtgact ggtaggaatt tgagcaagcc tgtcttgccg cgcgatgaga
     5581 aaggaarggc aagttacatt cagccccgta cctaggatca gtgctaattt ataacctcta
     5641 gccactcttt atgcagatcc gccgtgagkg tagcaaggca gctaaatcag ctggtgagaa
     5701 aggaaccata ccctgcaaga cctgtcgcaa cgcaaagggc aaaggtaagc tatccaagct
     5761 agtttgggac tagattctaa ctagtctcag caccgtgtgg ttcaaagcca tattgcgagt
     5821 tttggcgctt tttctcatcg attgacgagg caaagggagc gagtatgcaa cctcgtaagt
     5881 cagactcaga caaagaccaa ctagttattg accagtcttc tagaaggctc tgttgtggat
     5941 cttgaggccc tggagagttc ctccaacaat ccggagacaa gcaagtcctc gtcggactag
     6001 tcactaacta gactctaact agttgcagac atggataatg caaaagagac aagcaatgaa
     6061 gaaagtggta agacatttct cctttgtggt tctggactag tctttgacta gtcacagtct
     6121 taaacaagga aaatgagcat gaaaatgagg aggaaaaggc tgctgagccc gaggaagtgc
     6181 agggtgatgg cagacatggt aggttaatac cttgttagtt attgctagtc actgactagt
     6241 caataactag tctctgaaca ccttgcaatc actccgtttg cgcagctgaa cagtggtgag
     6301 gataatagta gtaagttatt ctagcttcag agttatagga gactagatac taactagtat
     6361 tagttgcaac taacctggat ctcagagact ttggcctcaa tctagaatct atctagttgt
     6421 caactagact gtggtatcat tgtcttttat tttcctagtc ctggaactag cttctaacta
     6481 gtctccctaa tatgtggctg tcttgttttt tttttttngt ttccctaccc ggatatctag
     6541 tccccttcta ggttctgtta acctctcggg ctctgattta gtttaacgca aacctgagat
     6601 tagtttctaa ctagtctcta ggttttctat ccacctttaa ttgtaataat aaatacaagc
     6661 aacgtttata cgtcaaaagc atttataaac ttttacccta aagtagcttg cttgtgtgtt
     6721 tagtttataa ttagtctctt attaatttga tgtaggtaag cccgccacaa atatatattt
     6781 ttaacaagat accgtggaaa aacttcgtgc tatcacaaaa cagtatacaa aaaataagct
     6841 taacaatcta ttctccgctt ggtgatgcta aagggctttc aatagacctt gtaagtgaag
     6901 gagatggagc cgtcaatccg ctaccctgcc tctggtcagt tggtctcagc aatgtaccct
     6961 gtgagtcttg atagactagt tggtgactag tctctgtaga tggaacaaat ggtgcttgag
     7021 agggcacgta tggcagagct ccagtcatct gcggaacata tactgggccc gggaagatct
     7081 catggtcata gctgtttccg ctgagggttt aattaagacc tcagctcctc gtgtactgtg
     7141 taagcgccca ctaggtaata tgacatgatt acgaattcga gctcggtacc cggccgggga
     7201 tcctctagac gtggagttac cagtgattga ccaatgtttt atcttctaca gttctgcctg
     7261 tctaccccat tctagctgta cctgactaca gagtagttta attgtggttg accccacagt
     7321 cggaggcgga ggaatacagc accgatgtgg cctgtctcca tccagattgg cacgcaattt
     7381 ttacacgcgg aaaagatcga gatagagtac gactttaaat ttagtccccg gcggcttcta
     7441 ttttagaata tttgagattt gattctcaag caattgattt ggttgggtca ccctcaattg
     7501 gataatatac ctcattgctc ggctacttca actcatcaat caccgtcata ccccgcatat
     7561 aaccctccat tcccacgatg tcgtccaagt cgcaattgac ttacggtgct cgagccagca
     7621 agcaccccaa tcctctggca aagagacttt ttgagattgc cgaagcaaag aagacaaacg
     7681 ttaccgtctc tgctgatgtg acgacaaccc gagaactcct ggacctcgct gaccgtacgg
     7741 aagctgttgg atccaataca tatgccgtct agcaatggac taatcaactt ttgatgatac
     7801 aggtctcggt ccctacatcg ccgtcatcaa gacacacatc gacatcctca ccgatttcag
     7861 cgtcgacact atcaatggcc tgaatgtgct ggctcaaaag tacaactttt tgatcttcga
     7921 ggaccgcaaa ttcatcgaca tcggcaatac cgtccagaag caataccacg gcggtgctct
     7981 gaggatctcc gaatgggccc acattatcaa ctgcagcgtt ctccctggcg agggcatcgt
     8041 cgaggctctg gcccagaccg catctgcgca agacttcccc tatggtcctg agagaggact
     8101 gttggtcctg gcagagatga cccccaaagg atcgctggct acgggcgagt ataccaaggc
     8161 atcggttgac tacgctcgca aatacaagaa cttcgttatg ggtttcgtgt cgacgcgggc
     8221 cctgacggaa gtgcagtcgg atgtgtcttc agcctcggag gatgaagatt tcgtggtctt
     8281 cacgacgggt gtgaacctct cttccaaagg agataagctt ggacagcaat accagactcc
     8341 tgcatcggct attggacgcg gtgccgactt tatcatcgcc ggtcgaggca tctacgctgc
     8401 tcccgacccg gttgaagctg cacagcggta ccagaaagaa ggctgggaag cttatatggc
     8461 cagagtatgc ggcaagtcat gatttcctct tggagcaaaa gtgtagtgcc agtacgagtg
     8521 ttgtggagga aggctgcata cattgtgcct gtcattaaac gatgagctcg tccgtattgg
     8581 cccctgtaat gccatgtttt ccgcccccaa tcgtcaaggt tttccctttg ttagattcct
     8641 accagtcatc tagcaaggcg gccgcagcta gcacaattga ggcatcccca ctaccgcatt
     8701 aagacctcag cgcggccgca aatttaaata aaatgaagtg aagttcctat actttctaga
     8761 gaataggaac ttctatagtg agtcgaataa gggcgacaca aaatttattc taaatgcata
     8821 ataaatactg ataacatctt atagtttgta ttatattttg tattatcgtt gacatgtata
     8881 attttgatat caaaaactga ttttcccttt attattttcg agatttattt tcttaattct
     8941 ctttaacaaa ctagaaatat tgtatataca aaaaatcata aataatagat gaatagttta
     9001 attataggtg ttcatcaatc gaaaaagcaa cgtatcttat ttaaagtgcg ttgctttttt
     9061 ctcatttata aggttaaata attctcatat atcaagcaaa gtgacaggcg cccttaaata
     9121 ttctgacaaa tgctctttcc ctaaactccc cccataaaaa aacccgccga agcgggtttt
     9181 tacgttattt gcggattaac gattactcgt tatcagaacc gcccaggggg cccgagctta
     9241 agactggccg tcgttttaca acacagaaag agtttgtaga aacgcaaaaa ggccatccgt
     9301 caggggcctt ctgcttagtt tgatgcctgg cagttcccta ctctcgcctt ccgcttcctc
     9361 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     9421 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     9481 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     9541 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     9601 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     9661 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     9721 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     9781 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     9841 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     9901 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtgggcta actacggcta
     9961 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
    10021 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
    10081 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
    10141 ggggtctgac gctcagtgga acgacgcgcg cgtaactcac gttaagggat tttggtcatg
    10201 agcttgcgcc gtcccgtcaa gtcagcgtaa tgctctgctt
//
