LOCUS       sequence-494102-        5049 bp    DNA     circular SYN 01-DEC-2025
DEFINITION  Modified plasmid from pADH100 to perform HIS-FLP CRISPR in 
            Candidozyma auris.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5049)
  AUTHORS   Sofras D, Carolus H, Subotic A, Romero CL, Ennis CL, Hernday AD, 
            Nobile CJ, Rybak JM, Van Dijck P
  TITLE     A comparative evaluation of CRISPR-Cas9 allele editing systems in 
            Candida auris: challenging research in a challenging bug.
  JOURNAL   bioRxiv [Preprint]. 2025 Jan 11:2025.01.09.632232. doi: 
            10.1101/2025.01.09.632232.
  PUBMED    39829791
REFERENCE   2  (bases 1 to 5049)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 1, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2025 Jan 11:2025.01.09.632232. doi: 
            10.1101/2025.01.09.632232."
COMMENT     Sequence Label: pADH100Cau
FEATURES             Location/Qualifiers
     source          1..5049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(124..915,916..984))
                     /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     747..766
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(985..1089)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1157..1175
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_RNA        2505..2580
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(2660..2693)
                     /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(4243..4260)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4414..5002)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4494..4513)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
       61 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
      121 cagttaccaa tgcttgatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
      181 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
      241 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
      301 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
      361 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
      421 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
      481 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
      541 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
      601 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
      661 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
      721 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
      781 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
      841 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
      901 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
      961 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     1021 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     1081 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     1141 gacattaacc tataaaaata ggcgtatcac gaggcccttt gacggcacgg ccacgcgttt
     1201 aaaccgccag gtgatgctga agctattgaa gctttggatg gttctttcac taccgatact
     1261 gtttttagag ttactgctac tggtgatggt ttcactttga gagaagttcc agttgatcca
     1321 ccattgacta aggttttccc agatgatgaa tccgatgatg aatccgatgc tggtgaagat
     1381 ggtgatccag attctagaac tttcgttgct tatggtgatg atggtgattt ggctggtttc
     1441 gttgttgttt cttattctgg ttggaacaga agattgactg ttgaagatat tgaagttgct
     1501 ccagaacata gaggtcatgg tgttggtaga gctttgatgg gtttggctac tgaattcgcc
     1561 agagaaagag gtgctggtca tttgtggttg gaagttacca atgttaatgc tccagctatt
     1621 catgcttata gaagaatggg tttcactttg tgtggtttgg atactgcttt atacgatggt
     1681 actgcttccg atggtgaaca agctttgtat atgtctatgc catgtccata ataactcgag
     1741 tcagtactga caataaaaag attcttgttt tcaagaactt gtcatttgta tagttttttt
     1801 atattgtagt tgttctattt taatcaaatg ttagcgtgat ttatattttt tttcgcctcg
     1861 acatcatctg cccagatgcg aagttaagtg cgcagaaagt aatatcatgc gtcaatcgta
     1921 tgtgaatgct acagactcaa tcaacgaagt tttttttttt tttttttttt ttttcttctt
     1981 catagacaat aaccttatca aagtttataa aactatatcg caatagaaaa caaaccccca
     2041 atcagcttaa cttttagagt cacttaccct acaaaggtgt aagcttccct cagatcaggc
     2101 ggcacgcatc tgatatctaa catccattgt aaaattagtg aagattaaca ctcagagcca
     2161 gttattttga acacagatgg cttttggctg ctcttatcgt ctacgtttct tttttttttt
     2221 tttttttttt tttttttttt cctaattttt tacgagttca ctaattggtt tcgggggctc
     2281 aattttcgaa tgcgacattc ataaactttg accttttatg tgttgatgcg gataattttg
     2341 acctcaaaac tcggaaaaat tgatgtatag cgtattcgac atggctttaa aagtcgcaga
     2401 gttctacgat tttcatttgc actggcccat ccggtaagcc tcggggctcc ggcatgtgag
     2461 ggttcgactc cctcagcaga aaacaggaga cggaattccg tctcgtttta gagctagaaa
     2521 tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc
     2581 tttttttctc gagttttttt atcgagtgtt taaggataat gataactgaa gagaagaatt
     2641 agttttgccg ccaccgcgtg aagttcctat actttctaga gaataggaac ttcccgccga
     2701 tcaaaatgtc tagagaagat cctattaagg ctgaaaaaga tttttctgaa actcttgaag
     2761 agcagatccc tctcattgag aagacttctg attacaaaca agctatcgag aagtacttgg
     2821 tgttggaaaa gcaaatccgt caatcttcag atttggctct gagtaagaag atcttaagca
     2881 aaattgttga tacgctagtc cataatgatg attgggacta cttggatgaa ttaatcacgc
     2941 ctttgtcaaa gaaacatggc caactcaaga gctccattca actgatgatt caagagatca
     3001 tcaagagcct cgaaacgtta gataacgaca agcctgtgca attagagaag aagatgaaga
     3061 ttattgaaac tattcgtacg gtgactgata agaagatctt tgtggaggtt gagagggcgg
     3121 tcgtttccaa gatgttggct gaaatatact tggagaagaa gaatgacttg gataaggccg
     3181 cagatatctt atgtgacttg caggtcgaga catactcgtt gatgcctttt gaagagaaaa
     3241 ttgagtacat tctcgagcaa attaggttga cattgcaaaa aaaagacttt gcacaagcaa
     3301 agattttgag cagaaagatt ctcttaaaaa cgttgaaaaa ttttgaaagg gccgacatat
     3361 ttaaggcaac gtacttgaag tacttactag agatcaacaa acatgagaat gactacatca
     3421 tggttgtcaa caactcactt ctcttgatag agatcccact agtcaaggaa tcttcagaat
     3481 acaaggactt gcttgtgtcc atcatctact acatcattct atcgccattc gataaccttc
     3541 aatccgattt gattcataaa atcaagctca acactgcttt ttctaagaat gtggattcta
     3601 aggtgttcaa gcttttggaa attttcacaa caaatgagct cattcactct gaaaatatcg
     3661 aaatggaata tcgtgaaaac tatcttaata agtcacctgt ttttgctgat aacgagacga
     3721 acaacacaaa cttacagaac agaattattg agcacaattt gcgtgtaatc aacatgtact
     3781 atctgtttat caagttagat cggttggctc atttgttaca agtcagttca caagatgcgg
     3841 aggcacatgt cagtggtatg gtaaacaaag gtatgatcac cgcgaaaatc aaccgtcccc
     3901 agggtattat caagttcgac aacttgaaac atggtactag tgccggcgct gctcagatta
     3961 ccagtaatag tgagaacatc aatgatttgt tgaacaactg gatatctgac gttgacaagc
     4021 tattggaaga ggtcgacagc attggtcact tgattaacaa ggaggaaatg atgtatggta
     4081 ttaagcaaaa cctgtgatat acataatgaa tagctcaaat cttaacagaa tttcttgacc
     4141 ctttacttac acaataatag caaaaatgaa agcacccaaa gaggcaaatc ctgaaagtgt
     4201 gctcatctga ttcgcgtcgt tacgtttgct gggtttaaac tcctcgctca ctgactcgct
     4261 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4321 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4381 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4441 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4501 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4561 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4621 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4681 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4741 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4801 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4861 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4921 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4981 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5041 gtggaacga
//
