LOCUS       sequence-180508-        3936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NAT-marked C. albicans ADE2-specific gRNA expression construct; part
            2 of 2 of C.alb HIS1-FLP CRISPR system. Use with pADH99 CAS9 
            expression construct..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3936)
  AUTHORS   Nguyen N, Quail MMF, Hernday AD
  TITLE     An Efficient, Rapid, and Recyclable System for CRISPR-Mediated 
            Genome Editing in Candida albicans.
  JOURNAL   mSphere. 2017 Apr 26;2(2):e00149-17. doi: 
            10.1128/mSphereDirect.00149-17. eCollection 2017 Mar-Apr.
  PUBMED    28497115
REFERENCE   2  (bases 1 to 3936)
  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"; doi: 
            "10.1128/mSphereDirect.00149-17"; journalName: "mSphere"; date: 
            "2017-04-26- 26"; volume: "2"; issue: "2"; pages: "e00149-17"
COMMENT     Sequence Label: pADH100-2
FEATURES             Location/Qualifiers
     source          1..3936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     4..22
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      593..790
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     misc_RNA        1114..1189
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(1216..1249)
                     /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)"
     promoter        complement(join(1794..1800,1801..1818,1819..1824))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1839..1860
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1977..1994)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2148..2736)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2228..2247)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2907..3698,3699..3767))
                     /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     3530..3549
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3768..3872)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaaaataggc gtatcacgag gccctttgac ggcacggcca cgcgtttaaa ccgccaggtg
       61 atgctgaagc tattgaagct ttggatggtt ctttcactac cgatactgtt tttagagtta
      121 ctgctactgg tgatggtttc actttgagag aagttccagt tgatccacca ttgactaagg
      181 ttttcccaga tgatgaatcc gatgatgaat ccgatgctgg tgaagatggt gatccagatt
      241 ctagaacttt cgttgcttat ggtgatgatg gtgatttggc tggtttcgtt gttgtttctt
      301 attctggttg gaacagaaga ttgactgttg aagatattga agttgctcca gaacatagag
      361 gtcatggtgt tggtagagct ttgatgggtt tggctactga attcgccaga gaaagaggtg
      421 ctggtcattt gtggttggaa gttaccaatg ttaatgctcc agctattcat gcttatagaa
      481 gaatgggttt cactttgtgt ggtttggata ctgctttata cgatggtact gcttccgatg
      541 gtgaacaagc tttgtatatg tctatgccat gtccataata actcgagtca gtactgacaa
      601 taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt
      661 tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc
      721 agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt
      781 cgctatactg ctgtcgattc gatactaaca tctaatcaac tcccagatca ccactataca
      841 tttaaaagtc attggtcaat aactatactc gagtattgcc tcatcaaaga aacaatcaaa
      901 tattatagat actcactcca tcacgtgata atttcactgg tatggaaaag tggaaaattt
      961 tataaaaaaa aatttgatgc ctttggcata gctgaaactt cggcccaata ggattggaga
     1021 atatgttttc gcagcgttct tacaattaaa ttgtggtgga agttcgagac ttgcgtaaac
     1081 tatttttaat ttgtctatag tacagatgcc aaggttttag agctagaaat agcaagttaa
     1141 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct ttttttctcg
     1201 agttttttta tcgaggaagt tcctatactt tctagagaat aggaacttcc caccttgatg
     1261 tacacacctt taaagtcaat aggcattctc gtttgtaatt ccttgacaca catatccatt
     1321 aactttaaac catcttcgac cgtcatgtcc tttctataat gtttatctaa caaagatgtt
     1381 gtgtaaaatg cagcgtagcc atgagcacca taagggagtt caacttgtgt acccaaataa
     1441 tctatccaat tcaacgatgg ctcatttttc tttacatcat atcctccaat taaacagttc
     1501 acctggtacg gttttctaga tctaagggaa ctggccaatt ggtttctcac aaacgaggct
     1561 gttgcctttg gagataactc aatgtcattt tctctcatcg agtacaattg aatgtttgct
     1621 tgaacatact cggcaaactg aaccgtatca ccagcttcac cagtgtatgc tattaagttg
     1681 tgtgcattta aatgtcttgt tttatcatct gtatccttta aaattgaaat acctcttgtg
     1741 gcagctttac ggtgtttaaa ccccagcgcc tggcgggcgc tcacaattcc acacaacata
     1801 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     1861 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     1921 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     1981 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2041 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2101 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2161 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2221 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2281 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2341 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2401 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2461 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2521 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2581 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2641 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2701 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2761 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2821 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2881 atatatgagt aaacttggtc tgacagttac caatgcttga tcagtgaggc acctatctca
     2941 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3001 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3061 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3121 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3181 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3241 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3301 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3361 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3421 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3481 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3541 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3601 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3661 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3721 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3781 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3841 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     3901 gtctaagaaa ccattattat catgacatta acctat
//
