LOCUS       sequence-180522-        4150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NAT-marked ""empty"" 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 4150)
  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 4150)
  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
FEATURES             Location/Qualifiers
     source          1..4150
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     111..130
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(349..453)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     521..539
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      1110..1307
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     misc_RNA        1629..1704
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(join(2525..2531,2532..2549,2550..2555))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2570..2591
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2708..2725)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2879..3467)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2959..2978)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3638..4150,1..279,280..348))
                     /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"
ORIGIN
        1 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
       61 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
      121 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
      181 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
      241 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
      301 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
      361 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
      421 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
      481 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
      541 ctttgacggc acggccacgc gtttaaaccg ccaggtgatg ctgaagctat tgaagctttg
      601 gatggttctt tcactaccga tactgttttt agagttactg ctactggtga tggtttcact
      661 ttgagagaag ttccagttga tccaccattg actaaggttt tcccagatga tgaatccgat
      721 gatgaatccg atgctggtga agatggtgat ccagattcta gaactttcgt tgcttatggt
      781 gatgatggtg atttggctgg tttcgttgtt gtttcttatt ctggttggaa cagaagattg
      841 actgttgaag atattgaagt tgctccagaa catagaggtc atggtgttgg tagagctttg
      901 atgggtttgg ctactgaatt cgccagagaa agaggtgctg gtcatttgtg gttggaagtt
      961 accaatgtta atgctccagc tattcatgct tatagaagaa tgggtttcac tttgtgtggt
     1021 ttggatactg ctttatacga tggtactgct tccgatggtg aacaagcttt gtatatgtct
     1081 atgccatgtc cataataact cgagtcagta ctgacaataa aaagattctt gttttcaaga
     1141 acttgtcatt tgtatagttt ttttatattg tagttgttct attttaatca aatgttagcg
     1201 tgatttatat tttttttcgc ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga
     1261 aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat
     1321 actaacatct aatcaactcc cagatcacca ctatacattt aaaagtcatt ggtcaataac
     1381 tatactcgag tattgcctca tcaaagaaac aatcaaatat tatagatact cactccatca
     1441 cgtgataatt tcactggtat ggaaaagtgg aaaattttat aaaaaaaaat ttgatgcctt
     1501 tggcatagct gaaacttcgg cccaatagga ttggagaata tgttttcgca gcgttcttac
     1561 aattaaattg tggtggaagt tcgagacttg cgtaaactat ttttaatttg gagacggaat
     1621 tccgtctcgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
     1681 gaaaaagtgg caccgagtcg gtgctttttt tctcgagttt ttttatcgag ttgagaccag
     1741 aagtcgttaa aggtacaaac ttgatcattg ttcgtgaatt agttggtggt atttatttcg
     1801 gtgatcgtga agaacaagaa gaaagtgaag ataaacaaac cgcatgggat actgaaaaat
     1861 acaccgtcga tgaagtcacc agaatcaccc gtatggctgc cttcatggct ttacaacata
     1921 acccaccatt accaatctgg tcattggata aggctaatgt tttagcttct tctagattat
     1981 ggagaagaac tgtcgataaa gtgatttctg aagagttccc aactttgtct gttcaacatc
     2041 aattgattga ctccgccgcc atgattttaa ttcaaaaccc aaccaaattg aatggtataa
     2101 tcatcacttc taacatgttt ggtgatatca tttccgatga agcttcagtt attccaggtt
     2161 ccttgggttt attaccatct gcatctttgg cttcattgcc agataccaac accgcttttg
     2221 gtctttatga accatgtcac ggttctgctc ctgatttacc agccaataaa gttaacccaa
     2281 ttgctactat tttatctgct gcttctatgt tgagattatc tttggattgt gtgaaagaag
     2341 ctgaagcttt agaagaagcc gtgaaacaag tcttggataa aggtatcaga actgcagatt
     2401 taagaggtac cagttcagct actgaagttg gtgacgcgat tgtcgaagct gttactaaaa
     2461 tcttaaaaga aaaagcttaa cggtgtttaa accccagcgc ctggcgggcg ctcacaattc
     2521 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     2581 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     2641 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2701 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2761 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2821 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2881 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2941 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3001 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3061 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3121 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3181 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3241 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3301 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3361 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3421 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3481 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3541 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3601 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgcttg atcagtgagg
     3661 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3721 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3781 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3841 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3901 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3961 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4021 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4081 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4141 attctcttac
//
