LOCUS       sequence-370106-        7795 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Codon optomized Kan resistance gene in the pSFS2A flipper backbone 
            for genetic manipulation of Candida albicans using G418 selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7795)
  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 7795)
  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: "mSphere. 
            2022 Aug 15:e0034722. doi: 10.1128/msphere.00347-22."
COMMENT     Sequence Label: pSFS2A-CaKan
FEATURES             Location/Qualifiers
     source          1..7795
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      5..461
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(92..111)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     302..323
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     587..609
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     601..618
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     602..618
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     628..647
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        628..646
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(676..709)
                     /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             1268..2539
                     /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
                     TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR
                     YPAWNGIISQEVLDYLSSYINRRI"
     protein_bind    complement(5081..5114)
                     /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(5116..5132)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5116..5132)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5169..5189)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5169..5187)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5208..5224)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5208..5224)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5221..5243)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5232..5248
                     /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(5256..5262,5263..5280,5281..5286))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5301..5322
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5439..5456)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5610..6198)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5690..5709)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        6418..6520
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             6521..7180
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(6587..6606)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(7669..7773)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
       61 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
      121 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
      181 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
      241 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
      301 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
      361 ttataaggga ttttgccgat ttcggcctat tggttaaaaa aatgagctga tttaacaaaa
      421 atttaacgcg aattttaaca aaatattaac gcttacaatt tccattcgcc attcaggctg
      481 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
      541 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
      601 tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga attgggtacc
      661 gggccccccc tcgaggaagt tcctatactt tctagagaat aggaacttcg gatccaataa
      721 tgattggttt gatatttttg tctagtacca tctgtaccat tacacttaaa ttatctttat
      781 atctgtctaa ctcgactgtc tggatttcat tgatgtagtc gtatgcatcg ttagttccaa
      841 aaaatattgt catcaatttg atattggttt ccgactctaa aatttttgga agaatttgtc
      901 tagcgtgctc tgagttgtag ccactgaaac cacggttaat aacatccaat tttcggatat
      961 acacattctg taatgctgga tgaaagccat actgggtaca actaaactgg gtgatggagt
     1021 caccgaacaa cacaaattta ccgtattcca tgattgctat ggttgagaat tttttttttt
     1081 tcttgtccca cgccattttt caaattatgc agttgagaat gttagttttt gtgtacaccc
     1141 cgttcgctga atatttcgga ataattcaaa gattggggag tgggggaggc gatagacgaa
     1201 gacacggtat aaaaatgggc aaaattttcc ccaacttttt gcagtggttt aactaataat
     1261 cgtcgacatg ccacaatttg atatattatg taaaacacca cctaaggtgc ttgttcgtca
     1321 gtttgtggaa aggtttgaaa gaccttcagg tgagaaaata gcattatgtg ctgctgaact
     1381 aacctattta tgttggatga ttacacataa cggaacagca atcaagagag ccacattcat
     1441 gagctataat actatcataa gcaattcgtt gagtttcgat attgtcaata aatcactcca
     1501 gtttaaatac aagacgcaaa aagcaacaat tttggaagcc tcattaaaga aattgattcc
     1561 tgcttgggaa tttacaatta ttccttacta tggacaaaaa catcaatctg atatcactga
     1621 tattgtaagt agtttgcaat tacagttcga atcatcggaa gaagcagata agggaaatag
     1681 ccacagtaaa aaaatgctta aagcacttct aagtgagggt gaaagcatct gggagatcac
     1741 tgagaaaata ctaaattcgt ttgagtatac ttcgagattt acaaaaacaa aaactttata
     1801 ccaattcctc ttcctagcta ctttcatcaa ttgtggaaga ttcagcgata ttaagaacgt
     1861 tgatccgaaa tcatttaaat tagtccaaaa taagtatttg ggagtaataa tccagtgttt
     1921 agtgacagag acaaagacaa gcgttagtag gcacatatac ttctttagcg caaggggtag
     1981 gatcgatcca cttgtatatt tggatgaatt tttgaggaat tctgaaccag tcctaaaacg
     2041 agtaaatagg accggcaatt cttcaagcaa taaacaggaa taccaattat taaaagataa
     2101 cttagtcaga tcgtacaata aagctttgaa gaaaaatgcg ccttattcaa tctttgctat
     2161 aaaaaatggc ccaaaatctc acattggaag acatttgatg acctcatttc tttcaatgaa
     2221 gggcctaacg gagttgacta atgttgtggg aaattggagc gataagcgtg cttctgccgt
     2281 ggccaggaca acgtatactc atcagataac agcaatacct gatcactact tcgcactagt
     2341 ttctcggtac tatgcatatg atccaatatc aaaggaaatg atagcattga aggatgagac
     2401 taatccaatt gaggagtggc agcatataga acagctaaag ggtagtgctg aaggaagcat
     2461 acgatacccc gcatggaatg ggataatatc acaggaggta ctagactacc tttcatccta
     2521 cataaataga cgcatataag agtgaaattc tggaaatctg gaaatctggt tttgtattct
     2581 tgttattctt ctttttgtta ttacatatat aacttgttac ttttttaaaa aaatctttgt
     2641 atattttata aatatataaa actaaattta agaaaaagag aaaaatgttt tatttgagag
     2701 attgaaattt tacttgaatt tagcttagct tttataaagt attattatgt aaaaaaacaa
     2761 aacaaatata cattaaaaag ttaagactat aaaatagcca cccaaggcat ttctatatct
     2821 tgttgttgtt gttttcatct tctgtatcag aggaacttat tttattattt tcgtcacggg
     2881 tattttctct tgtttgatga ttcatcccat tcattccatc ataaaatgtc gagcgtcaaa
     2941 actagagaat aataaagaaa acgatctttt caaaaagaaa aaacctttta gttttccttt
     3001 gttgttgttg tgggtgtgtg ctatttatat tatatagttt actcataata ccataaaata
     3061 ttcggtttga ttaggttatt ttaataagct aatttgtttc taatcgtgta atttatgctg
     3121 tgtatattaa gtagtgtgtg cactgcccaa aaatgtttgt tgtttatagt cggttaaaga
     3181 gaaaaaagaa aaaaagatcc atacacacac gttaattagt tgttcaacgt aatacactca
     3241 tattttgttc ttatttgctt tcggtcgctg ttctcaccaa gatttattgc caacgaaaca
     3301 attttttttt atatattttc agatttttct ttttttcctt tcctttcctt ttctaatttt
     3361 cactcctggt tttctttctt tcttagaaac attatctcga tattaatatt aaaaaaatat
     3421 aatcattcaa aatggacggt ggtatgtttt agtttagctt caattctaat tgattgatta
     3481 atcagttgat tggtttcaat atgacaaatg ggtagggtgg gaaaacttca ttttcaattc
     3541 agatcaaact tttttgttgt cgacataata tttctcgttt gggatgttac tgtcacatta
     3601 ataatacaca cacatcagct tataattttg aaagtaattt atcagatatg ttgtgacgat
     3661 cagtggaaat ggctaacttc aatgtatctg ttcttcccct ttttcaaagt tcacgttttt
     3721 tgattgattg attgatctgt cggcagtggt ttcaaaacca ttcggtgagt aatcctatca
     3781 atcaatgtta cgacaaaagg ctcaatattc aaaattgcaa tgttttatgt tttcctacgt
     3841 gtacttgtgc aaggcaattg attcaacatt gcttttggtg tttgacgagt ttctagtttg
     3901 gacttgtgtt gttatctggg ctatacagat ttcccggctc actatgaatt ttttttttcg
     3961 acgctcagtg cacacaacta taaacaacac aaacacaaac acagcaagaa aaaaaaaaaa
     4021 cgaacattga attgaaacca agccaactga aaaattcctt atttaaatga ctgtcatact
     4081 aacccatttt tatagaagaa gttgctgctt tagttatcga taacggttct catatgggaa
     4141 aagaaaagac ccacgtgtct cgaccacgtt tgaactcaaa tatggacgca gacttatacg
     4201 ggtacaaatg ggctcgtgac aacgtcgggc aatccggggc taccatatac agattgtatg
     4261 ggaaacctga tgctccagag ttgttcttaa agcatggtaa aggatctgtc gcaaacgatg
     4321 taaccgatga gatggtcaga cttaactggc ttacagagtt tatgcctttg ccaactatta
     4381 agcacttcat tagaacccct gacgatgcat ggttattgac tactgctatc ccaggtaaga
     4441 ctgccttcca agtcttagag gaatatcctg acagtggaga gaacatcgtc gatgcattgg
     4501 ccgtcttttt acgtcgtctt cattccatcc ctgtttgcaa ttgccctttt aactccgata
     4561 gagtctttcg tttggctcag gcccaatctc gtatgaataa cgggttggtg gacgcttcag
     4621 attttgatga tgaacgtaac ggttggcctg tagaacaagt gtggaaagaa atgcacaaac
     4681 ttttaccatt ttctccagat tctgtcgtga ctcatggaga cttcagttta gataatctta
     4741 tattcgatga agggaaattg atcggttgca tagatgtagg gagagtgggt attgccgatc
     4801 gttatcaaga tttggcaatc ttgtggaact gccttggaga gttctcccct agtttgcaaa
     4861 agcgtttatt ccagaagtac ggaatagata atccagatat gaataaattg cagttccatt
     4921 tgatgttaga tgagttcttc tgacatatgt gaagtgtgaa gggggagatt ttcactttat
     4981 tagatttgta tatatgtata ataaataaat aaataagtta aataaataat tagataaggg
     5041 tggtaattat tactatttac aatcaaaggt ggtcctgcag gaagttccta tactttctag
     5101 agaataggaa cttcagatcc actagttcta gagcggccgc caccgcggtg gagctccagc
     5161 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     5221 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5281 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5341 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5401 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5461 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5521 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5581 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5641 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5701 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5761 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5821 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5881 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5941 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6001 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6061 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6121 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6181 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6241 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6301 atccttttcg accgaataaa tacctgtgac ggaagatcac ttcgcagaat aaataaatcc
     6361 tggtgtccct gttgataccg ggaagccctg ggccaacttt tggcgaaaat gagacgttga
     6421 tcggcacgta agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt
     6481 ttttgagttg tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg
     6541 atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc
     6601 agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac
     6661 cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat
     6721 gaatgctcat ccggaattac gtatggcaat gaaagacggt gagctggtga tatgggatag
     6781 tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag
     6841 tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta
     6901 cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc
     6961 caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt
     7021 cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct
     7081 ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga
     7141 attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg
     7201 gtgcccttaa acgcctggtt gctacgcctg aataagtgat aataagcgga tgaatggcag
     7261 aaattcgaaa gcaaattcga cccggtcgtc ggttcagggc agggtcgtta aatagccgct
     7321 tatgtctatt gctggtttac cggtttattg actaccggaa gcagtgtgac cgtgtgcttc
     7381 tcaaatgcct gaggccagtt tgctcaggct ctccccgtgg aggtaataat tgacgatatg
     7441 atcctttttt tctgatcaaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7501 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7561 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7621 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7681 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7741 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgcca
//
