LOCUS       sequence-180523-        4152 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NAT-marked C. albicans ADE2-specific gRNA expression construct; part
            2 of 2 of C.mal LEUpOUT CRISPR system. Use with pADH140 CAS9 
            expression construct..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4152)
  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 4152)
  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: pADH143-2
FEATURES             Location/Qualifiers
     source          1..4152
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     138..156
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      727..924
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     misc_RNA        1248..1323
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(join(2144..2150,2151..2168,2169..2174))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2189..2210
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2327..2344)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2498..3086)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2578..2597)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3257..4048,4049..4117))
                     /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     3880..3899
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4118..4152,1..70))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
       61 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
      121 tgacattaac ctataaaaat aggcgtatca cgaggccctt tgacggcacg gccacgcgtt
      181 taaaccgcca ggtgatgctg aagctattga agctttggat ggttctttca ctaccgatac
      241 tgtttttaga gttactgcta ctggtgatgg tttcactttg agagaagttc cagttgatcc
      301 accattgact aaggttttcc cagatgatga atccgatgat gaatccgatg ctggtgaaga
      361 tggtgatcca gattctagaa ctttcgttgc ttatggtgat gatggtgatt tggctggttt
      421 cgttgttgtt tcttattctg gttggaacag aagattgact gttgaagata ttgaagttgc
      481 tccagaacat agaggtcatg gtgttggtag agctttgatg ggtttggcta ctgaattcgc
      541 cagagaaaga ggtgctggtc atttgtggtt ggaagttacc aatgttaatg ctccagctat
      601 tcatgcttat agaagaatgg gtttcacttt gtgtggtttg gatactgctt tatacgatgg
      661 tactgcttcc gatggtgaac aagctttgta tatgtctatg ccatgtccat aataactcga
      721 gtcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt
      781 tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc
      841 gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt
      901 atgtgaatgc tggtcgctat actgctgtcg attcgatact aacatctaat caactcccag
      961 atcaccacta tacatttaaa agtcattggt caataactat actcgagtat tgcctcatca
     1021 aagaaacaat caaatattat agatactcac tccatcacgt gataatttca ctggtatgga
     1081 aaagtggaaa attttataaa aaaaaatttg atgcctttgg catagctgaa acttcggccc
     1141 aataggattg gagaatatgt tttcgcagcg ttcttacaat taaattgtgg tggaagttcg
     1201 agacttgcgt aaactatttt taatttgtct atagtacaga tgccaaggtt ttagagctag
     1261 aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg
     1321 tgcttttttt ctcgagtttt tttatcgagt tgagaccaga agtcgttaaa ggtacaaact
     1381 tgatcattgt tcgtgaatta gttggtggta tttatttcgg tgatcgtgaa gaacaagaag
     1441 aaagtgaaga taaacaaacc gcatgggata ctgaaaaata caccgtcgat gaagtcacca
     1501 gaatcacccg tatggctgcc ttcatggctt tacaacataa cccaccatta ccaatctggt
     1561 cattggataa ggctaatgtt ttagcttctt ctagattatg gagaagaact gtcgataaag
     1621 tgatttctga agagttccca actttgtctg ttcaacatca attgattgac tccgccgcca
     1681 tgattttaat tcaaaaccca accaaattga atggtataat catcacttct aacatgtttg
     1741 gtgatatcat ttccgatgaa gcttcagtta ttccaggttc cttgggttta ttaccatctg
     1801 catctttggc ttcattgcca gataccaaca ccgcttttgg tctttatgaa ccatgtcacg
     1861 gttctgctcc tgatttacca gccaataaag ttaacccaat tgctactatt ttatctgctg
     1921 cttctatgtt gagattatct ttggattgtg tgaaagaagc tgaagcttta gaagaagccg
     1981 tgaaacaagt cttggataaa ggtatcagaa ctgcagattt aagaggtacc agttcagcta
     2041 ctgaagttgg tgacgcgatt gtcgaagctg ttactaaaat cttaaaagaa aaagcttaac
     2101 ggtgtttaaa ccccagcgcc tggcgggcgc tcacaattcc acacaacata cgagccggaa
     2161 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     2221 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     2281 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     2341 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2401 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2461 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2521 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2581 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2641 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     2701 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     2761 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     2821 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     2881 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     2941 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3001 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3061 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3121 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3181 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3241 aaacttggtc tgacagttac caatgcttga tcagtgaggc acctatctca gcgatctgtc
     3301 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3361 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3421 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3481 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3541 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     3601 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     3661 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     3721 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     3781 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     3841 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     3901 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     3961 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4021 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4081 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4141 gaagcattta tc
//
