LOCUS       sequence-337337-        4075 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses sgRNA (ENO1 gRNA) in Candida auris..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4075)
  AUTHORS   Santana DJ, O'Meara TR
  TITLE     Forward and reverse genetic dissection of morphogenesis identifies 
            filament-competent Candida auris strains.
  JOURNAL   Nat Commun. 2021 Dec 10;12(1):7197. doi: 10.1038/s41467-021-27545-5.
  PUBMED    34893621
REFERENCE   2  (bases 1 to 4075)
  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"; journalName: "Nat 
            Commun."; date: "2021-12-10"; volume: "12(1):7197. doi"; pages: " 
            10.1038/s41467-021-27545-5"
COMMENT     Sequence Label: pTO136_pADH1-tRNA-Ala-gRNA-tracrRNA-HDV-tAgTEF2
FEATURES             Location/Qualifiers
     source          1..4075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        983..1058
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           1063..1130
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     terminator      1145..1342
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1423..1439)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1423..1439)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1436..1458)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1447..1463
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1471..1477,1478..1495,1496..1501))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1516..1537
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1654..1671)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1825..2413)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1905..1924)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2584..3375,3376..3444))
                     /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     3207..3226
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3445..3549)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3617..3635
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3673..3695)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3795..3814
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4008..4030
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4022..4039
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4023..4039
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gctctaaagt ttccaatacg atttaaaacg cataaggtgg tccggcagca ggcccttcaa
       61 agcgggaagt atacttctag aaactattca ttgcgctaga cgctcatatg caacgccact
      121 caaatctgca aaattattca gatctatgct gacatactct gacgattctg tggcgatgca
      181 cttttttttt tttttttttt catttttttt tttatttttt ttgcaaccgt tccattcgac
      241 aggttacatt tctagaaacg tctgtcccca acatggcacg gaaaaattag accagattcc
      301 cacgtactgg gcgacttggc tgtgcatgat gcagtttcgg gcatcatgat ccttaagcgc
      361 atacatcagg tgatgcgaca ccacatcgtg acgaaaggaa gctggagcca ggccaatttc
      421 accgcagcct ttgaccctga gccgcaagac accacccctc cttggcggcg atgcccggtt
      481 cttccaattg cggctcgtag gtgcagttga aacgacgatc tgatgttcca tgaaagagca
      541 gatagaaaaa agtgagattg gcgaagacag taaaatttca acaccacatg tcaatcttac
      601 aaaaacaatc aatcagttca tttttcttct cccctcaact acaattttca aaaatgccag
      661 tgaatgaaaa aaaagaccca ttgtcccaaa actggctccc ttcttgaaac cacctgattc
      721 ctcgagcgaa tcggatggga aaaagccgca ccgtcaccca atgcgaacca ccggccatgc
      781 gtcagatggc accctttggc tgcgaaaaat ataaaagcag agcaacaccg gtggaatttc
      841 caaagtcttt tcttctccat catcaatcaa tcttcacgaa atcctactcg ggcgtgtggc
      901 gtagttggta gcgcgttcgc ttagcatgcg aaaggtctcc ggttcgaatc cggactcgtc
      961 cagtttgcgc ttcaaaccac ttgttttaga gctagaaata gcaagttaaa ataaggctag
     1021 tccgttatca acttgaaaaa gtggcaccga gtcggtgctt ttggccggca tggtcccagc
     1081 ctcctcgctg gcgccggctg ggcaacatgc ttcggcatgg cgaatgggac taactcgagt
     1141 cagtactgac aataaaaaga ttcttgtttt caagaacttg tcatttgtat agttttttta
     1201 tattgtagtt gttctatttt aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga
     1261 catcatctgc ccagatgcga agttaagtgc gcagaaagta atatcatgcg tcaatcgtat
     1321 gtgaatgctg gtcgctatac tgctgtcgat tcgatactaa cgccgccatc cagtggatcc
     1381 tctagagtcg acctgcaggc atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt
     1441 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     1501 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     1561 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     1621 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     1681 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     1741 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1801 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1861 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1921 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1981 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2041 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2101 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2161 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2221 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     2281 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2341 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2401 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2461 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     2521 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     2581 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     2641 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     2701 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     2761 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     2821 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     2881 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     2941 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3001 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     3061 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     3121 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     3181 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     3241 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     3301 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     3361 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     3421 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     3481 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     3541 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     3601 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     3661 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     3721 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     3781 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     3841 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     3901 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     3961 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     4021 ttgtaaaacg acggccagtg aattcgagct cggtacccga gatagatcga aatac
//
