LOCUS       sequence-415594-       10347 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AMA1 plasmid with amdS selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10347)
  AUTHORS   Vanegas KG, Rendsvig JKH, Jarczynska ZD, Cortes MVCB, van Esch AP, 
            Morera-Gomez M, Contesini FJ, Mortensen UH
  TITLE     A Mad7 System for Genetic Engineering of Filamentous Fungi.
  JOURNAL   J Fungi (Basel). 2022 Dec 22;9(1):16. doi: 10.3390/jof9010016.
  PUBMED    36675838
REFERENCE   2  (bases 1 to 10347)
  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: "J Fungi 
            (Basel)."; date: "2022-12-22"; volume: "9(1):16. doi"; pages: " 
            10.3390/jof9010016"
COMMENT     Sequence Label: pDIV297
FEATURES             Location/Qualifiers
     source          1..10347
                     /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)"
     CDS             7467..9113
                     /codon_start=1
                     /gene="amdS"
                     /product="acetamidase from Aspergillus nidulans"
                     /label=amdS
                     /note="confers ability to process acetamide for selection 
                     in fungal cells"
                     /translation="MPQSWEELAADKRARLAKTIPDEWKVQTLPAEDSVIDFPKKSGIL
                     SEAELKITEASAADLVSKLAAGELTSVEVTLAFCKRAAIAQQLTNCAHEFFPDAALAQA
                     RELDEYYAKHKRPVGPLHGLPISLKDQLRVKGYETSMGYISWLNKYDEGDSVLTTMLRK
                     AGAVFYVKTSVPQTLMVCETVNNIIGRTVNPRNKNWSCGGSSGGEGAIVGIRGGVIGVG
                     TDIGGSIRVPAAFNFLYGLRPSHGRLPYAKMANSMEGQETVHSVVGPITHSVEDLRLFT
                     KSVLGQEPWKYDSKVIPMPWRQSESDIIASKIKNGGLNIGYYNFDGNVLPHPPILRGVE
                     TTVAALAKAGHTVTPWTPYKHDFGHDLISHIYAADGSADVMRDISASGEPAIPNIKDLL
                     NPNIKAVNMNELWDTHLQKWNYQMEYLEKWREAEEKAGKELDAIIAPITPTAAVRHDQF
                     RYYGYASVINLLDFTSVVVPVTFADKNIDKKNESFKAVSELDALVQEEYDPEAYHGAPV
                     AVQVIGRRLSEERTLAIAEEVGKLLGNVVTP"
     rep_origin      complement(9636..10224)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9716..9735)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10346..10347,1..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 tggtcctccg 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 atatgggtca
      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 gtcyctgaga 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 gcttkgatag cttacctttg ccctttgcgt
     2701 tgcgacaggt cttgcagggt atggttcctt tctcaccagc tgatttagct gccttgctac
     2761 cctcacggcg 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 ggacttgcct ttggcctcca
     3301 tcagttcctt catagtggga ggtccattgt gcaatgtaaa ctccatgccg tgggagttct
     3361 tgtccttcaa gtgcttgacc aatatgtttc tgttggcaga gggaacctgt caactagtta
     3421 ataactagtc agaaactagt atagcagtag actcactgta cgcttgaggc atcccttcac
     3481 tcggcagtag acttcatatg 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 aactagttat gtacctggag tttggatata ggaatgtgtt gtggtaacga aatgtaaggg
     3901 ggaggaaaga aaaagtcggt caagaggtaa ctctaagtcg gccattcctt tttgggaggc
     3961 gctaaccata aacggcatgg tcgacttaga gttagctcag ggaatttagg gagttatctg
     4021 cgaccaccga ggaacggcgg 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 ggcaagtcct 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 gccgtgaggg tagcaaggca gctaaatcag ctggtgagaa
     5701 aggaaccata ccctgcaaga cctgtcgcaa cgcaaagggc aaaggtaagc tatcmaagct
     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 ctcagrgact 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 gctggctgag ggtttaatgc tgagggttta attaagacct cagcgctagt
     7141 ggaggtcaac acatcaatgc ctattttggt ttagtcgtcc aggcggtgag cacaaaattt
     7201 gtgtcgtttg acaagatggt tcatttaggc aactggtcag atcagcccca cttgtagcag
     7261 tagcggcggc gctcgaagtg tgactcttat tagcagacag gaacgaggac attattatca
     7321 tctgctgctt ggtgcacgat aacttggtgc gtttgtcaag caaggtaagt ggacgacccg
     7381 gtcatacctt cttaagttcg cccttcctcc ctttatttca gattcaatct gacttaccta
     7441 ttctacccaa gcatccaaaa acaaccatgc ctcaatcctg ggaagaactg gccgctgata
     7501 agcgcgcccg cctcgcaaaa accatccctg atgaatggaa agtccagacg ctgcctgcgg
     7561 aagacagcgt tattgatttc ccaaagaaat cggggatcct ttcagaggcc gaactgaaga
     7621 tcacagaggc ctccgctgca gaccttgtgt ccaagctggc ggccggagag ttgacctcgg
     7681 tggaagttac gctagcattc tgtaaacggg cagcaatcgc ccagcagtta acaaactgcg
     7741 cccacgagtt cttccctgac gccgctctcg cgcaggcaag ggaactcgat gaatactacg
     7801 caaagcacaa gagacccgtt ggtccactcc atggcctccc catctctctc aaagaccagc
     7861 ttcgagtcaa gggctacgaa acatcaatgg gctacatctc atggctaaac aagtacgacg
     7921 aaggggactc ggttctgaca accatgctcc gcaaagccgg tgccgtcttc tacgtcaaga
     7981 cctctgtccc gcagaccctg atggtctgcg agacagtcaa caacatcatc gggcgcaccg
     8041 tcaacccacg caacaagaac tggtcgtgcg gcggcagttc tggtggtgag ggtgcgatcg
     8101 ttgggattcg tggtggcgtc atcggtgtag gaacggatat cggtggctcg attcgagtgc
     8161 cggccgcgtt caacttcctg tacggtctaa ggccgagtca tgggcggctg ccgtatgcaa
     8221 agatggcgaa cagcatggag ggtcaggaga cggtgcacag cgttgtcggg ccgattacgc
     8281 actctgttga ggacctccgc ctcttcacca aatccgtcct cggtcaggag ccatggaaat
     8341 acgattccaa ggtcatcccc atgccctggc gccagtccga gtcggacatt attgcctcca
     8401 agatcaagaa cggcgggctc aatatcggct actacaactt cgacggcaat gtccttccac
     8461 accctcctat cctgcgcggc gtggaaacca ccgtcgccgc actcgccaaa gccggtcaca
     8521 ccgtgacccc gtggacgcca tacaagcacg atttcggcca cgatctcatc tcccatatct
     8581 acgcggctga cggcagcgcc gacgtaatgc gcgatatcag tgcatccggc gagccggcga
     8641 ttccaaatat caaagaccta ctgaacccga acatcaaagc tgttaacatg aacgagctct
     8701 gggacacgca tctccagaag tggaattacc agatggagta ccttgagaaa tggcgggagg
     8761 ctgaagaaaa ggccgggaag gaactggacg ccatcatcgc gccgattacg cctaccgctg
     8821 cggtacggca tgaccagttc cggtactatg ggtatgcctc tgtgatcaac ctgctggatt
     8881 tcacgagcgt ggttgttccg gttacctttg cggataagaa catcgataag aagaatgaga
     8941 gtttcaaggc ggttagtgag cttgatgccc tcgtgcagga agagtatgat ccggaggcgt
     9001 accatggggc accggttgca gtgcaggtta tcggacggag actcagtgaa gagaggacgt
     9061 tggcgattgc agaggaagtg gggaagttgc tgggaaatgt ggtgactcca tagatcagta
     9121 ctccgacgcc gaccaacacc gccggtccga cgcggcccga cgggtccgag gagcttgaga
     9181 tccacttaac gttactgaaa tcatcaaaca gcttgacgaa tctggatata agatcgttgg
     9241 tgtcgatgtc agctccggag ttgagacaaa tggtgttcag gatctcgata agatacgttc
     9301 atttgtccaa gcagcaaaga gtgccttcta gtgatttaat agctccatgt caacaagaat
     9361 aaaacgcgtt tcgggtttac ctcttccaga tacagctcat ctgcaatgca ttaatgcatt
     9421 ggacctcgca accctagtac gcccttcagg ctccggcgaa gcagaagaat agcttagcag
     9481 agtctatttt cattttcggg agacgagatc aagcagatca acggtcgtca agagacctac
     9541 gagactgagg aatccgctct tggctcatta agacctcagc gagcaaaagg ccagcaaaag
     9601 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     9661 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     9721 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     9781 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     9841 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     9901 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     9961 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
    10021 gtaggcggtg ctacagagtt cttgaagtgg tgggctaact acggctacac tagaagaaca
    10081 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
    10141 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
    10201 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
    10261 cagtggaacg acgcgcgcgt aactcacgtt aagggatttt ggtcatgagc ttgcgccgtc
    10321 ccgtcaagtc agcgtaatgc tctgctt
//
