LOCUS       sequence-180511-        4150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NAT-marked ""empty"" 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 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: pADH118
FEATURES             Location/Qualifiers
     source          1..4150
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     38..56
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      627..824
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     misc_RNA        1146..1221
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(join(2042..2048,2049..2066,2067..2072))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2087..2108
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2225..2242)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2396..2984)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2476..2495)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3155..3946,3947..4015))
                     /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     3778..3797
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4016..4120)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
       61 tgacggcacg gccacgcgtt taaaccgcca ggtgatgctg aagctattga agctttggat
      121 ggttctttca ctaccgatac tgtttttaga gttactgcta ctggtgatgg tttcactttg
      181 agagaagttc cagttgatcc accattgact aaggttttcc cagatgatga atccgatgat
      241 gaatccgatg ctggtgaaga tggtgatcca gattctagaa ctttcgttgc ttatggtgat
      301 gatggtgatt tggctggttt cgttgttgtt tcttattctg gttggaacag aagattgact
      361 gttgaagata ttgaagttgc tccagaacat agaggtcatg gtgttggtag agctttgatg
      421 ggtttggcta ctgaattcgc cagagaaaga ggtgctggtc atttgtggtt ggaagttacc
      481 aatgttaatg ctccagctat tcatgcttat agaagaatgg gtttcacttt gtgtggtttg
      541 gatactgctt tatacgatgg tactgcttcc gatggtgaac aagctttgta tatgtctatg
      601 ccatgtccat aataactcga gtcagtactg acaataaaaa gattcttgtt ttcaagaact
      661 tgtcatttgt atagtttttt tatattgtag ttgttctatt ttaatcaaat gttagcgtga
      721 tttatatttt ttttcgcctc gacatcatct gcccagatgc gaagttaagt gcgcagaaag
      781 taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg attcgatact
      841 aacatctaat caactcccag atcaccacta tacatttaaa agtcattggt caataactat
      901 actcgagtat tgcctcatca aagaaacaat caaatattat agatactcac tccatcacgt
      961 gataatttca ctggtatgga aaagtggaaa attttataaa aaaaaatttg atgcctttgg
     1021 catagctgaa acttcggccc aataggattg gagaatatgt tttcgcagcg ttcttacaat
     1081 taaattgtgg tggaagttcg agacttgcgt aaactatttt taatttggag acggaattcc
     1141 gtctcgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa
     1201 aaagtggcac cgagtcggtg ctttttttct cgagtttttt tatcgagtta aaagctgaag
     1261 tggtcaaagg aaccaattta attattgttc gtgaattagt tggtggtata tattttggtg
     1321 aacgtcaaga acaagaagaa agtgaagaca aaaaaacagc ttgggataca gaaaagtata
     1381 ctgttgacga agttactcgt atcactcgta tggcagcatt tatggctttg cagcacaatc
     1441 caccattgcc aatatggtct ttggacaaag ctaatgtttt ggcatcgtcg agattgtgga
     1501 gaaagaccgt tgataaagtg atttctgaag aatttccagc tttgtctgtg cagcatcaat
     1561 tgattgattc ggctgcaatg attttgatcc aaaacccaac caaattgaac ggtatcatca
     1621 ttacatctaa catgtttggt gatatcatct cagatgaggc atccgttatc ccaggatctt
     1681 tgggtttatt gccatctgct tcattggctt cattgccaga taccaacact gcttttggtc
     1741 tttatgagcc ttgccatggg tctgctccag atttaccagc caacaaagtg aatccaattg
     1801 ctacaatctt gtctgccgct tccatgttga gattatcctt ggattgtgtt aaggaagctg
     1861 aagcattgga agaagctgtc aaacaggttt tggatagcgg tatcagaaca gcagatttaa
     1921 gaggtactag ctctactaca gaagtcggtg acgcaatagc cgaaacagtc actaaaattt
     1981 taaaacaaaa tacataacgg tgtttaaacc ccagcgcctg gcgggcgctc acaattccac
     2041 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     2101 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     2161 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     2221 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2281 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2341 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2401 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2461 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2521 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2581 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2641 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2701 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2761 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2821 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2881 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2941 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3001 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3061 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3121 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttgatc agtgaggcac
     3181 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3241 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3301 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3361 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3421 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3481 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3541 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3601 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3661 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3721 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3781 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     3841 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     3901 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     3961 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4021 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4081 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4141 cacctgacgt
//
