LOCUS       sequence-180512-        4152 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NAT-marked C. albicans ADE2-specific gRNA expression construct; part
            2 of 2 of C.alb LEUpOUT CRISPR system. Use with pADH137 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: pADH118-2
FEATURES             Location/Qualifiers
     source          1..4152
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     72..90
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      661..858
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     misc_RNA        1182..1257
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(join(2078..2084,2085..2102,2103..2108))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2123..2144
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2261..2278)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2432..3020)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2512..2531)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3191..3982,3983..4051))
                     /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     3814..3833
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4052..4152,1..4))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
       61 taacctataa aaataggcgt atcacgaggc cctttgacgg cacggccacg cgtttaaacc
      121 gccaggtgat gctgaagcta ttgaagcttt ggatggttct ttcactaccg atactgtttt
      181 tagagttact gctactggtg atggtttcac tttgagagaa gttccagttg atccaccatt
      241 gactaaggtt ttcccagatg atgaatccga tgatgaatcc gatgctggtg aagatggtga
      301 tccagattct agaactttcg ttgcttatgg tgatgatggt gatttggctg gtttcgttgt
      361 tgtttcttat tctggttgga acagaagatt gactgttgaa gatattgaag ttgctccaga
      421 acatagaggt catggtgttg gtagagcttt gatgggtttg gctactgaat tcgccagaga
      481 aagaggtgct ggtcatttgt ggttggaagt taccaatgtt aatgctccag ctattcatgc
      541 ttatagaaga atgggtttca ctttgtgtgg tttggatact gctttatacg atggtactgc
      601 ttccgatggt gaacaagctt tgtatatgtc tatgccatgt ccataataac tcgagtcagt
      661 actgacaata aaaagattct tgttttcaag aacttgtcat ttgtatagtt tttttatatt
      721 gtagttgttc tattttaatc aaatgttagc gtgatttata ttttttttcg cctcgacatc
      781 atctgcccag atgcgaagtt aagtgcgcag aaagtaatat catgcgtcaa tcgtatgtga
      841 atgctggtcg ctatactgct gtcgattcga tactaacatc taatcaactc ccagatcacc
      901 actatacatt taaaagtcat tggtcaataa ctatactcga gtattgcctc atcaaagaaa
      961 caatcaaata ttatagatac tcactccatc acgtgataat ttcactggta tggaaaagtg
     1021 gaaaatttta taaaaaaaaa tttgatgcct ttggcatagc tgaaacttcg gcccaatagg
     1081 attggagaat atgttttcgc agcgttctta caattaaatt gtggtggaag ttcgagactt
     1141 gcgtaaacta tttttaattt gtctatagta cagatgccaa ggttttagag ctagaaatag
     1201 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt
     1261 ttttctcgag tttttttatc gagttaaaag ctgaagtggt caaaggaacc aatttaatta
     1321 ttgttcgtga attagttggt ggtatatatt ttggtgaacg tcaagaacaa gaagaaagtg
     1381 aagacaaaaa aacagcttgg gatacagaaa agtatactgt tgacgaagtt actcgtatca
     1441 ctcgtatggc agcatttatg gctttgcagc acaatccacc attgccaata tggtctttgg
     1501 acaaagctaa tgttttggca tcgtcgagat tgtggagaaa gaccgttgat aaagtgattt
     1561 ctgaagaatt tccagctttg tctgtgcagc atcaattgat tgattcggct gcaatgattt
     1621 tgatccaaaa cccaaccaaa ttgaacggta tcatcattac atctaacatg tttggtgata
     1681 tcatctcaga tgaggcatcc gttatcccag gatctttggg tttattgcca tctgcttcat
     1741 tggcttcatt gccagatacc aacactgctt ttggtcttta tgagccttgc catgggtctg
     1801 ctccagattt accagccaac aaagtgaatc caattgctac aatcttgtct gccgcttcca
     1861 tgttgagatt atccttggat tgtgttaagg aagctgaagc attggaagaa gctgtcaaac
     1921 aggttttgga tagcggtatc agaacagcag atttaagagg tactagctct actacagaag
     1981 tcggtgacgc aatagccgaa acagtcacta aaattttaaa acaaaataca taacggtgtt
     2041 taaaccccag cgcctggcgg gcgctcacaa ttccacacaa catacgagcc ggaagcataa
     2101 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     2161 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2221 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2281 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2341 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2401 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2461 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2521 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2581 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2641 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2701 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2761 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2821 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2881 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2941 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3001 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3061 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3121 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3181 ggtctgacag ttaccaatgc ttgatcagtg aggcacctat ctcagcgatc tgtctatttc
     3241 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3301 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3361 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3421 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3481 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3541 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3601 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3661 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3721 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3781 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     3841 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3901 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3961 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4021 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4081 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4141 aaataggggt tc
//
