LOCUS       sequence-349717-        4321 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C. auris LEUpOUT HYG marker gRNA expression construct. Use with 
            pCE38 CAS9 expression construct..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4321)
  AUTHORS   Ennis CL, Hernday AD, Nobile CJ
  TITLE     A Markerless CRISPR-Mediated System for Genome Editing in Candida 
            auris Reveals a Conserved Role for Cas5 in the Caspofungin Response.
  JOURNAL   Microbiol Spectr. 2021 Dec 22;9(3):e0182021. doi: 
            10.1128/Spectrum.01820-21. Epub 2021 Nov 3.
  PUBMED    34730409
REFERENCE   2  (bases 1 to 4321)
  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: "Microbiol 
            Spectr."; date: "2021-12-22"; volume: "9(3):e0182021. doi"; pages: "
            10.1128/Spectrum.01820-21. Epub 2021 Nov 3"
COMMENT     Sequence Label: pCE41
FEATURES             Location/Qualifiers
     source          1..4321
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(72..863,864..932))
                     /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     695..714
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(933..1037)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1105..1123
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_RNA        2384..2459
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(join(3280..3286,3287..3304,3305..3310))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3325..3346
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3463..3480)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3634..4222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3714..3733)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
       61 tggtctgaca gttaccaatg cttgatcagt gaggcaccta tctcagcgat ctgtctattt
      121 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
      181 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
      241 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
      301 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
      361 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
      421 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
      481 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
      541 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
      601 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
      661 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
      721 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
      781 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
      841 actttcacca gagtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
      901 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
      961 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     1021 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     1081 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttga cggcacggcc
     1141 acgcgtttaa accgcctact gtataagtcg aagagcacaa ggggttactt tacaagactt
     1201 gccagaaacc gaattaccag ccgtgcttca gccagtagct gaggctatgg acgcaatcgc
     1261 tgcagccgat ttaagtcaaa catcaggttt cggtcctttt gggccacaag ggatcggtca
     1321 atatactacc tggcgtgatt ttatttgcgc catagcagat cctcacgtgt accactggca
     1381 aaccgtcatg gatgacacag tctcagcatc cgtggcccag gcacttgacg aattgatgtt
     1441 atgggcagag gattgtcctg aggtgcgaca cttggttcat gccgactttg ggagtaacaa
     1501 cgtcttaaca gacaacggac gaataaccgc tgtgattgac tggtcagaag ctatgttcgg
     1561 tgactctcaa tacgaagttg ccaacatctt cttctggcga ccttggcttg cctgcatgga
     1621 acagcaaaca cgatacttcg aacgacgaca tccagaactt gccggatctc ctcgattacg
     1681 tgcctatatg ttacgtatcg gtttagacca attgtatcag agtttggtcg acggaaattt
     1741 cgacgacgcc gcatgggcac agggtcgttg tgacgccata gttagaagtg gagcaggtac
     1801 cgttgggaga actcagattg ctcgtagatc cgcagcagta tggacagatg gatgtgtaga
     1861 ggttttagcc gacagtggaa accgaagacc ttcaacaaga ccacgtgcca aggagtaaag
     1921 cttaactttt agagtcactt accctacaaa ggtgtaagct tccctcagat caggcggcac
     1981 gcatctgata tctaacatcc attgtaaaat tagtgaagat taacactcag agccagttat
     2041 tttgaacaca gatggctttt ggctgctctt atcgtctacg tttctttttt tttttttttt
     2101 tttttttttt ttttttttcc taatttttta cgagttcact aattggtttc gggggctcaa
     2161 ttttcgaatg cgacattcat aaactttgac cttttatgtg ttgatctgga taattttgac
     2221 ctcaaaactc ggaaaaattg atgtatagcg tattcgacat ggctttaaaa gtcgcagagt
     2281 tctacgattt tcatttgcac tggcccatcc ggtaagcctc ggggctccgg catgtgaggg
     2341 ttcgactccc tcagcagaaa acatgccgtg aaagaaaaca ccagttttag agctagaaat
     2401 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
     2461 ttttttctcg agttttttta tcgagtctcc attgaaggcc agtgttgtca aaggaaccaa
     2521 tttcaccgtg gtgcgtgaat tggtcggtgg tatctacttt ggtgagagac aagaacagga
     2581 agagtctgcc gacggtcaga gcgcttggga caccgagaag tacagtgtct ctgaagttga
     2641 aagaatcacc agaatggctg ccttcatggc tttgcagcac aaccctcctt tgcccatctg
     2701 gtccttagac aaggccaacg tgcttgcctc atctagattg tggagaaaga cggtcgacaa
     2761 ggtgatgagc gaggagttcc ctcaattgac catcaaccac cagctcattg actctgctgc
     2821 catgattttg gtgcagagcc caactaaatt gaacggtgtt gtgatcacgt caaacatgtt
     2881 tggtgatatt atttccgacg aggcttccgt gatccctggc tccttgggtc ttttaccttc
     2941 tgcatctttg gcttcattac cagatacaaa taaggcgttt ggtttgtacg aaccatgcca
     3001 cggttcggct ccggacttgc cagaaaacaa ggtcaaccct gtcgccacga ttttgtcggc
     3061 cgctatgatg ttgcgcttgt cattagactg cctcagagag gcagaggcct tggagaaggc
     3121 tgttcgtaac gttttggact ctggtattcg aaccgcagat ttgaagggct caagttccac
     3181 tcaggaagtg ggcgatgcca ttgctgctga ggtaaagaag cttttgcaga aataacggtg
     3241 tttaaacccc agcgcctggc gggcgctcac aattccacac aacatacgag ccggaagcat
     3301 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3361 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3421 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3481 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3541 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3601 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3661 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3721 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3781 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3841 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3901 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3961 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4021 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4081 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4141 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4201 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4261 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4321 c
//
