LOCUS       sequence-492801-        7663 bp    DNA     circular SYN 14-NOV-2025
DEFINITION  CaHygB hygromycin resistance gene in the pSFS2A flipper backbone for
            genetic manipulation of Candida albicans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7663)
  AUTHORS   Park SO, Frazer C, Bennett RJ
  TITLE     An Adjuvant-Based Approach Enables the Use of Dominant HYG and KAN 
            Selectable Markers in Candida albicans.
  JOURNAL   mSphere. 2022 Aug 15:e0034722. doi: 10.1128/msphere.00347-22.
  PUBMED    35968963
REFERENCE   2  (bases 1 to 7663)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "mSphere. 
            2022 Aug 15:e0034722. doi: 10.1128/msphere.00347-22."
COMMENT     Sequence Label: pSFS2A-CaHygB
FEATURES             Location/Qualifiers
     source          1..7663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     58..80
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     72..89
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     73..89
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..118
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        99..117
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(147..180)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             739..2010
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=FLP
                     /note="FLP is a site-specific recombinase from 
                     Saccharomyces cerevisiae. Recombination occurs at FRT 
                     sequences."
                     /translation="MPQFDILCKTPPKVLVRQFVERFERPSGEKIALCAAELTYLCWMI
                     THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI
                     IPYYGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN
                     SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET
                     KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR
                     SYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR
                     TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQPKGSAEGSMR
                     YPAWNGIISQEVLDYLSSYINRRI"
     CDS             2967..3992
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     protein_bind    complement(4420..4453)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(4455..4471)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4455..4471)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4508..4528)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4508..4526)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4547..4563)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4547..4563)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4560..4582)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4571..4587
                     /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(4595..4601,4602..4619,4620..4625))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4640..4661
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4778..4795)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4949..5537)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5029..5048)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5757..5859
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5860..6519
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5926..5945)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(7008..7112)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7139..7595
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7226..7245)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7436..7457
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
       61 agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg
      121 aattgggtac cgggcccccc ctcgaggaag ttcctatact ttctagagaa taggaacttc
      181 ggatccaata atgattggtt tgatattttt gtctagtacc atctgtacca ttacacttaa
      241 attatcttta tatctgtcta actcgactgt ctggatttca ttgatgtagt cgtatgcatc
      301 gttagttcca aaaaatattg tcatcaattt gatattggtt tccgactcta aaatttttgg
      361 aagaatttgt ctagcgtgct ctgagttgta gccactgaaa ccacggttaa taacatccaa
      421 ttttcggata tacacattct gtaatgctgg atgaaagcca tactgggtac aactaaactg
      481 ggtgatggag tcaccgaaca acacaaattt accgtattcc atgattgcta tggttgagaa
      541 tttttttttt ttcttgtccc acgccatttt tcaaattatg cagttgagaa tgttagtttt
      601 tgtgtacacc ccgttcgctg aatatttcgg aataattcaa agattgggga gtgggggagg
      661 cgatagacga agacacggta taaaaatggg caaaattttc cccaactttt tgcagtggtt
      721 taactaataa tcgtcgacat gccacaattt gatatattat gtaaaacacc acctaaggtg
      781 cttgttcgtc agtttgtgga aaggtttgaa agaccttcag gtgagaaaat agcattatgt
      841 gctgctgaac taacctattt atgttggatg attacacata acggaacagc aatcaagaga
      901 gccacattca tgagctataa tactatcata agcaattcgt tgagtttcga tattgtcaat
      961 aaatcactcc agtttaaata caagacgcaa aaagcaacaa ttttggaagc ctcattaaag
     1021 aaattgattc ctgcttggga atttacaatt attccttact atggacaaaa acatcaatct
     1081 gatatcactg atattgtaag tagtttgcaa ttacagttcg aatcatcgga agaagcagat
     1141 aagggaaata gccacagtaa aaaaatgctt aaagcacttc taagtgaggg tgaaagcatc
     1201 tgggagatca ctgagaaaat actaaattcg tttgagtata cttcgagatt tacaaaaaca
     1261 aaaactttat accaattcct cttcctagct actttcatca attgtggaag attcagcgat
     1321 attaagaacg ttgatccgaa atcatttaaa ttagtccaaa ataagtattt gggagtaata
     1381 atccagtgtt tagtgacaga gacaaagaca agcgttagta ggcacatata cttctttagc
     1441 gcaaggggta ggatcgatcc acttgtatat ttggatgaat ttttgaggaa ttctgaacca
     1501 gtcctaaaac gagtaaatag gaccggcaat tcttcaagca ataaacagga ataccaatta
     1561 ttaaaagata acttagtcag atcgtacaat aaagctttga agaaaaatgc gccttattca
     1621 atctttgcta taaaaaatgg cccaaaatct cacattggaa gacatttgat gacctcattt
     1681 ctttcaatga agggcctaac ggagttgact aatgttgtgg gaaattggag cgataagcgt
     1741 gcttctgccg tggccaggac aacgtatact catcagataa cagcaatacc tgatcactac
     1801 ttcgcactag tttctcggta ctatgcatat gatccaatat caaaggaaat gatagcattg
     1861 aaggatgaga ctaatccaat tgaggagtgg cagcatatag aacagccaaa gggtagtgct
     1921 gaaggaagca tgcgataccc cgcatggaat gggataatat cacaggaggt actagactac
     1981 ctttcatcct acataaatag acgcatataa gagtgaaatt ctggaaatct ggaaatctgg
     2041 ttttgtattc ttgttattct tctttttgtt attacatata taacttgtta cttttttaaa
     2101 aaaatctttg tatattttat aaatatataa aactaaattt aagaaaaaga gaaaaatgtt
     2161 ttatttgaga gattgaaatt ttacttgaat ttagcttagc ttttataaag tattattatg
     2221 taaaaaaaca aaacaaatat acattaaaaa gttaagacta taaaatagcc acccaaggca
     2281 tttctatccg ctgctaggcg cgccgtgcca cctgacgtcg tatagtgctt gctgttcgat
     2341 attgctagag gcaaagaaaa aggattacct cacgcctaaa atggaatttg aaaattgagg
     2401 agatgagttg agcttttgtc ttttacactt tccctattca attgtgccgc aaaaacttca
     2461 atatccaaca cagtgtgaaa tatttgttat gtataccgaa agccaaatac catttaaact
     2521 ggtatttcaa gatttcataa atcaagtttc tggttaaatt aattgctgtt aaacagaaca
     2581 agagcttcac attgatgttg gaataactac tatagaatta taaaatacac aaaaccaggt
     2641 cgtgtacgca ataacttttc aatgtatgaa aaaatttttt tttttttttg cttcttctct
     2701 tcgcttcttt tttctagtta tcaatttggc aatgaattga tttgattgcc ctttgtttag
     2761 tttactatga aaaaaaattt ttttttgtct ctgtcagatt ttctctcttt tttttttgcc
     2821 tcataatgaa caactcaaaa aagaatataa atttctcatc aaagaccttt ccaattgaaa
     2881 attttttctt tccttctttt cctttttttt tttctttctt tcttcttttc taagtataca
     2941 ttttatataa tcaatcctcg atcgagatga aaaaaccaga attgactgct acttctgttg
     3001 aaaaattttt gattgaaaaa tttgattctg tttctgattt gatgcaattg tctgaaggtg
     3061 aagaatcaag agctttttct tttgatgttg gtggtagagg ttatgttttg agagttaatt
     3121 cttgtgctga tggtttttat aaagatagat atgtttatag acattttgct tctgctgctt
     3181 tgccaattcc agaagttttg gatattggtg aattttctga atctttgact tattgtattt
     3241 caagaagagc gcaaggtgtt actttgcaag atttgccaga aactgaattg ccagctgttt
     3301 tgcaaccagt tgctgaagct atggatgcta ttgctgctgc tgatttgtct caaacttctg
     3361 gttttggtcc atttggtcca caaggtattg gtcaatatac tacttggaga gattttattt
     3421 gtgctattgc tgatccacat gtttatcatt ggcaaactgt tatggatgat actgtttctg
     3481 cttctgttgc tcaagcattg gatgaattga tgttgtgggc tgaagattgt ccagaagtta
     3541 gacatttggt tcatgctgat tttggttcta ataatgtttt gactgataat ggtagaatta
     3601 ctgctgttat tgattggtct gaagctatgt ttggtgattc tcaatatgaa gttgctaata
     3661 tttttttttg gagaccatgg ttggcatgta tggaacaaca aactagatat tttgaaagaa
     3721 gacatccaga attggctggt tctccaagat tgagagctta tatgttgaga attggtttgg
     3781 atcaattgta tcaatctttg gttgatggta attttgatga tgctgcttgg gctcaaggta
     3841 gatgtgatgc tattgttaga tcaggtgctg gtactgttgg tagaactcaa attgctagaa
     3901 gatcagctgc tgtttggact gatggttgtg ttgaagtttt ggctgattct ggtaatagaa
     3961 gaccatctac tagaccaaga gctaaagaat aaggatccga gtgaaattct ggaaatctgg
     4021 aaatctggtt ttgtattctt gttattcttc tttttgttat tacatatata acttgttact
     4081 tttttaaaaa aatctttgtt tattttataa atatataaaa ctaaatttaa gaaaaagaga
     4141 aaaatgtttt atttgagaga ttgaaatttt acttgaattt agcttagctt ttataaagta
     4201 ttattatgta aaaaaacaaa acaaatatac attaaaaagt taagactata aaatagccac
     4261 ccaaggcatt tctatatctt gttgttgttg ttttcatctt ctgtatcaga ggaacttatt
     4321 ttattatttt cgtcacgggt attttctctt gtttgatgat tcatcccatt cattccatca
     4381 taaaatgacg tctaagaaac cattattatc atgctgcagg aagttcctat actttctaga
     4441 gaataggaac ttcagatcca ctagttctag agcggccgcc accgcggtgg agctccagct
     4501 tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc
     4561 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4621 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4681 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4741 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4801 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4861 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4921 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4981 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5041 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5101 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5161 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5221 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5281 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5341 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5401 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5461 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5521 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5581 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5641 tccttttcga ccgaataaat acctgtgacg gaagatcact tcgcagaata aataaatcct
     5701 ggtgtccctg ttgataccgg gaagccctgg gccaactttt ggcgaaaatg agacgttgat
     5761 cggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt
     5821 tttgagttgt cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
     5881 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
     5941 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
     6001 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
     6061 aatgctcatc cggaattacg tatggcaatg aaagacggtg agctggtgat atgggatagt
     6121 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
     6181 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
     6241 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc
     6301 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
     6361 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
     6421 gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa
     6481 ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttttttaagg cagttattgg
     6541 tgcccttaaa cgcctggttg ctacgcctga ataagtgata ataagcggat gaatggcaga
     6601 aattcgaaag caaattcgac ccggtcgtcg gttcagggca gggtcgttaa atagccgctt
     6661 atgtctattg ctggtttacc ggtttattga ctaccggaag cagtgtgacc gtgtgcttct
     6721 caaatgcctg aggccagttt gctcaggctc tccccgtgga ggtaataatt gacgatatga
     6781 tccttttttt ctgatcaaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6841 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6901 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6961 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7021 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7081 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7141 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     7201 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     7261 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     7321 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     7381 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     7441 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     7501 gggattttgc cgatttcggc ctattggtta aaaaaatgag ctgatttaac aaaaatttaa
     7561 cgcgaatttt aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac
     7621 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gcc
//
