LOCUS       sequence-180507-        3934 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NAT-marked ""empty"" 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 3934)
  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 3934)
  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
FEATURES             Location/Qualifiers
     source          1..3934
                     /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        1112..1187
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(1214..1247)
                     /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(1792..1798,1799..1816,1817..1822))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1837..1858
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1975..1992)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2146..2734)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2226..2245)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2905..3696,3697..3765))
                     /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     3528..3547
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3766..3870)
                     /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 ttggagacgg aattccgtct cgttttagag ctagaaatag caagttaaaa
     1141 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttctcgag
     1201 tttttttatc gaggaagttc ctatactttc tagagaatag gaacttccca ccttgatgta
     1261 cacaccttta aagtcaatag gcattctcgt ttgtaattcc ttgacacaca tatccattaa
     1321 ctttaaacca tcttcgaccg tcatgtcctt tctataatgt ttatctaaca aagatgttgt
     1381 gtaaaatgca gcgtagccat gagcaccata agggagttca acttgtgtac ccaaataatc
     1441 tatccaattc aacgatggct catttttctt tacatcatat cctccaatta aacagttcac
     1501 ctggtacggt tttctagatc taagggaact ggccaattgg tttctcacaa acgaggctgt
     1561 tgcctttgga gataactcaa tgtcattttc tctcatcgag tacaattgaa tgtttgcttg
     1621 aacatactcg gcaaactgaa ccgtatcacc agcttcacca gtgtatgcta ttaagttgtg
     1681 tgcatttaaa tgtcttgttt tatcatctgt atcctttaaa attgaaatac ctcttgtggc
     1741 agctttacgg tgtttaaacc ccagcgcctg gcgggcgctc acaattccac acaacatacg
     1801 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     1861 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     1921 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     1981 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2041 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2101 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2161 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2221 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2281 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2341 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2401 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2461 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2521 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2581 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2641 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2701 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2761 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     2821 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     2881 atatgagtaa acttggtctg acagttacca atgcttgatc agtgaggcac ctatctcagc
     2941 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3001 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3061 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3121 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3181 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3241 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3301 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3361 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3421 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     3481 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     3541 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     3601 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     3661 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     3721 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     3781 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     3841 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     3901 ctaagaaacc attattatca tgacattaac ctat
//
