LOCUS       sequence-180525-        4152 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NAT-marked C. albicans URA3-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: pADH118-17
FEATURES             Location/Qualifiers
     source          1..4152
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(239..1030,1031..1099))
                     /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     862..881
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1100..1204)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1272..1290
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      1861..2058
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     misc_RNA        2382..2457
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(join(3278..3284,3285..3302,3303..3308))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3323..3344
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3461..3478)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(3632..4152,1..68))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3712..3731)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
       61 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
      121 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
      181 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
      241 accaatgctt gatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
      301 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
      361 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
      421 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
      481 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
      541 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
      601 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
      661 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
      721 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
      781 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
      841 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
      901 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
      961 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     1021 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     1081 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     1141 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     1201 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     1261 taacctataa aaataggcgt atcacgaggc cctttgacgg cacggccacg cgtttaaacc
     1321 gccaggtgat gctgaagcta ttgaagcttt ggatggttct ttcactaccg atactgtttt
     1381 tagagttact gctactggtg atggtttcac tttgagagaa gttccagttg atccaccatt
     1441 gactaaggtt ttcccagatg atgaatccga tgatgaatcc gatgctggtg aagatggtga
     1501 tccagattct agaactttcg ttgcttatgg tgatgatggt gatttggctg gtttcgttgt
     1561 tgtttcttat tctggttgga acagaagatt gactgttgaa gatattgaag ttgctccaga
     1621 acatagaggt catggtgttg gtagagcttt gatgggtttg gctactgaat tcgccagaga
     1681 aagaggtgct ggtcatttgt ggttggaagt taccaatgtt aatgctccag ctattcatgc
     1741 ttatagaaga atgggtttca ctttgtgtgg tttggatact gctttatacg atggtactgc
     1801 ttccgatggt gaacaagctt tgtatatgtc tatgccatgt ccataataac tcgagtcagt
     1861 actgacaata aaaagattct tgttttcaag aacttgtcat ttgtatagtt tttttatatt
     1921 gtagttgttc tattttaatc aaatgttagc gtgatttata ttttttttcg cctcgacatc
     1981 atctgcccag atgcgaagtt aagtgcgcag aaagtaatat catgcgtcaa tcgtatgtga
     2041 atgctggtcg ctatactgct gtcgattcga tactaacatc taatcaactc ccagatcacc
     2101 actatacatt taaaagtcat tggtcaataa ctatactcga gtattgcctc atcaaagaaa
     2161 caatcaaata ttatagatac tcactccatc acgtgataat ttcactggta tggaaaagtg
     2221 gaaaatttta taaaaaaaaa tttgatgcct ttggcatagc tgaaacttcg gcccaatagg
     2281 attggagaat atgttttcgc agcgttctta caattaaatt gtggtggaag ttcgagactt
     2341 gcgtaaacta tttttaattt ggtagttgaa ggattaaaac agttttagag ctagaaatag
     2401 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt
     2461 ttttctcgag tttttttatc gagttaaaag ctgaagtggt caaaggaacc aatttaatta
     2521 ttgttcgtga attagttggt ggtatatatt ttggtgaacg tcaagaacaa gaagaaagtg
     2581 aagacaaaaa aacagcttgg gatacagaaa agtatactgt tgacgaagtt actcgtatca
     2641 ctcgtatggc agcatttatg gctttgcagc acaatccacc attgccaata tggtctttgg
     2701 acaaagctaa tgttttggca tcgtcgagat tgtggagaaa gaccgttgat aaagtgattt
     2761 ctgaagaatt tccagctttg tctgtgcagc atcaattgat tgattcggct gcaatgattt
     2821 tgatccaaaa cccaaccaaa ttgaacggta tcatcattac atctaacatg tttggtgata
     2881 tcatctcaga tgaggcatcc gttatcccag gatctttggg tttattgcca tctgcttcat
     2941 tggcttcatt gccagatacc aacactgctt ttggtcttta tgagccttgc catgggtctg
     3001 ctccagattt accagccaac aaagtgaatc caattgctac aatcttgtct gccgcttcca
     3061 tgttgagatt atccttggat tgtgttaagg aagctgaagc attggaagaa gctgtcaaac
     3121 aggttttgga tagcggtatc agaacagcag atttaagagg tactagctct actacagaag
     3181 tcggtgacgc aatagccgaa acagtcacta aaattttaaa acaaaataca taacggtgtt
     3241 taaaccccag cgcctggcgg gcgctcacaa ttccacacaa catacgagcc ggaagcataa
     3301 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     3361 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     3421 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     3481 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3541 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3601 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3661 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3721 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3781 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3841 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3901 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3961 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4021 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4081 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4141 ccggcaaaca aa
//
