LOCUS       sequence-243299-        8239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-terminal fusions of C. albicans optimized mNeonGreen fluorescent 
            protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8239)
  AUTHORS   Frazer C, Hernday AD, Bennett RJ
  TITLE     Monitoring Phenotypic Switching in Candida albicans and the Use of 
            Next-Gen Fluorescence Reporters.
  JOURNAL   Curr Protoc Microbiol. 2019 Feb 12:e76. doi: 10.1002/cpmc.76.
  PUBMED    30747494
REFERENCE   2  (bases 1 to 8239)
  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: "Curr Protoc
            Microbiol. 2019 Feb 12:e76. doi: 10.1002/cpmc.76."
COMMENT     Sequence Label: pRB895
FEATURES             Location/Qualifiers
     source          1..8239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(109..130)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     321..340
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        454..558
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(1047..1706)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1621..1640
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1707..1809)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      2029..2617
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2518..2537
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2771..2788
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2905..2926
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2941..2946,2947..2964,2965..2971)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2979..2995
                     /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)."
     primer_bind     2984..3006
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3003..3019
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3003..3019
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3038..3058
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3040..3058
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     3095..3111
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3095..3111
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    3113..3146
                     /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             complement(5403..6674)
                     /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    7233..7266
                     /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(8024..8043)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8025..8043)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8053..8070)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8053..8069)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8062..8084)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(join(8210..8239,1..427))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 tcgcgttaaa tttttgttaa atcagctcat ttttttaacc aataggccga aatcggcaaa
       61 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
      121 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
      181 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
      241 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
      301 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
      361 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
      421 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
      481 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
      541 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
      601 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
      661 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
      721 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcactttttg atcagaaaaa
      781 aaggatcata tcgtcaatta ttacctccac ggggagagcc tgagcaaact ggcctcaggc
      841 atttgagaag cacacggtca cactgcttcc ggtagtcaat aaaccggtaa accagcaata
      901 gacataagcg gctatttaac gaccctgccc tgaaccgacg accgggtcga atttgctttc
      961 gaatttctgc cattcatccg cttattatca cttattcagg cgtagcaacc aggcgtttaa
     1021 gggcaccaat aactgcctta aaaaaattac gccccgccct gccactcatc gcagtactgt
     1081 tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga
     1141 atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg
     1201 ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag
     1261 ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt
     1321 tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg
     1381 tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg
     1441 tgaacactat cccatatcac cagctcaccg tctttcattg ccatacgtaa ttccggatga
     1501 gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc
     1561 tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga
     1621 gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg
     1681 gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgacaac
     1741 tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg
     1801 tgccgatcaa cgtctcattt tcgccaaaag ttggcccagg gcttcccggt atcaacaggg
     1861 acaccaggat ttatttattc tgcgaagtga tcttccgtca caggtattta ttcggtcgaa
     1921 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     1981 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2041 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2101 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2161 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2221 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2281 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2341 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2401 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2461 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2521 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2581 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2641 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     2701 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     2761 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     2821 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     2881 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     2941 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     3001 cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga
     3061 acaaaagctg gagctccacc gcggtggcgg ccgctctaga actagtggat ctgaagttcc
     3121 tattctctag aaagtatagg aacttcctgc aggaccacct ttgattgtaa atagtaataa
     3181 ttaccaccct tatctaatta tttatttaac ttatttattt atttattata catatataca
     3241 aatctaataa agtgaaaatc tcccccttca cacttcacat atgttaggcg tcatcctgtg
     3301 ctcccgagaa ccagtaccag tacatcgctg tttcgttcga gacttgaggt ctagttttat
     3361 acgtgaagag gtcaatgccg ccgagagtaa agccacattt tgcgtacaaa ttgcaggcag
     3421 gtacattgtt cgtttgtgtc tctaatcgta tgccaaggag ctgtctgctt agtgcccact
     3481 ttttcgcaaa ttcgatgaga ctgtgcgcga ctcctttgcc tcggtgcgtg tgcgacacaa
     3541 caatgtgttc gatagaggct agatcgttcc atgttgagtt gagttcaatc ttcccgacaa
     3601 gctcttggtc gatgaatgcg ccatagcaag cagagtcttc atcagagtca tcatccgaga
     3661 tgtaatcctt ccggtagggg ctcacacttc tggtagatag ttcaaagcct tggtcggata
     3721 ggtgcacatc gaacacttca cgaacaatga aatggttctc agcatccaat gtttccgcca
     3781 cctgctcagg gatcaccgaa attttcatat gagaaccgtt atcgataact aaagcagcaa
     3841 cttcttctat aaaaatgggt tagtatgaca gtcatttaaa taaggaattt ttcagttggc
     3901 ttggtttcaa ttcaatgttc gttttttttt tttcttgctg tgtttgtgtt tgtgttgttt
     3961 atagttgtgt gcactgagcg tcgaaaaaaa aaattcatag tgagccggga aatctgtata
     4021 gcccagataa caacacaagt ccaaactaga aactcgtcaa acaccaaaag caatgttgaa
     4081 tcaattgcct tgcacaagta cacgtaggaa aacataaaac attgcaattt tgaatattga
     4141 gccttttgtc gtaacattga ttgataggat tactcaccga atggttttga aaccactgcc
     4201 gacagatcaa tcaatcaatc aaaaaacgtg aactttgaaa aaggggaaga acagatacat
     4261 tgaagttagc catttccact gatcgtcaca acatatctga taaattactt tcaaaattat
     4321 aagctgatgt gtgtgtatta ttaatgtgac agtaacatcc caaacgagaa atattatgtc
     4381 gacaacaaaa aagtttgatc tgaattgaaa atgaagtttt cccaccctac ccatttgtca
     4441 tattgaaacc aatcaactga ttaatcaatc aattagaatt gaagctaaac taaaacatac
     4501 caccgtccat tttgaatgat tatatttttt taatattaat atcgagataa tgtttctaag
     4561 aaagaaagaa aaccaggagt gaaaattaga aaaggaaagg aaaggaaaaa aagaaaaatc
     4621 tgaaaatata taaaaaaaaa ttgtttcgtt ggcaataaat cttggtgaga acagcgaccg
     4681 aaagcaaata agaacaaaat atgagtgtat tacgttgaac aactaattaa cgtgtgtgta
     4741 tggatctttt tttctttttt ctctttaacc gactataaac aacaaacatt tttgggcagt
     4801 gcacacacta cttaatatac acagcataaa ttacacgatt agaaacaaat tagcttatta
     4861 aaataaccta atcaaaccga atattttatg gtattatgag taaactatat aatataaata
     4921 gcacacaccc acaacaacaa caaaggaaaa ctaaaaggtt ttttcttttt gaaaagatcg
     4981 ttttctttat tattctctag ttttgacgct cgacatttta tgatggaatg aatgggatga
     5041 atcatcaaac aagagaaaat acccgtgacg aaaataataa aataagttcc tctgatacag
     5101 aagatgaaaa caacaacaac aagatataga aatgccttgg gtggctattt tatagtctta
     5161 actttttaat gtatatttgt tttgtttttt tacataataa tactttataa aagctaagct
     5221 aaattcaagt aaaatttcaa tctctcaaat aaaacatttt tctctttttc ttaaatttag
     5281 ttttatatat ttataaaata tacaaagatt tttttaaaaa agtaacaagt tatatatgta
     5341 ataacaaaaa gaagaataac aagaatacaa aaccagattt ccagatttcc agaatttcac
     5401 tcttatatgc gtctatttat gtaggatgaa aggtagtcta gtacctcctg tgatattatc
     5461 ccattccatg cggggtatcg tatgcttcct tcagcactac cctttagctg ttctatatgc
     5521 tgccactcct caattggatt agtctcatcc ttcaatgcta tcatttcctt tgatattgga
     5581 tcatatgcat agtaccgaga aactagtgcg aagtagtgat caggtattgc tgttatctga
     5641 tgagtatacg ttgtcctggc cacggcagaa gcacgcttat cgctccaatt tcccacaaca
     5701 ttagtcaact ccgttaggcc cttcattgaa agaaatgagg tcatcaaatg tcttccaatg
     5761 tgagattttg ggccattttt tatagcaaag attgaataag gcgcattttt cttcaaagct
     5821 ttattgtacg atctgactaa gttatctttt aataattggt attcctgttt attgcttgaa
     5881 gaattgccgg tcctatttac tcgttttagg actggttcag aattcctcaa aaattcatcc
     5941 aaatatacaa gtggatcgat cctacccctt gcgctaaaga agtatatgtg cctactaacg
     6001 cttgtctttg tctctgtcac taaacactgg attattactc ccaaatactt attttggact
     6061 aatttaaatg atttcggatc aacgttctta atatcgctga atcttccaca attgatgaaa
     6121 gtagctagga agaggaattg gtataaagtt tttgtttttg taaatctcga agtatactca
     6181 aacgaattta gtattttctc agtgatctcc cagatgcttt caccctcact tagaagtgct
     6241 ttaagcattt ttttactgtg gctatttccc ttatctgctt cttccgatga ttcgaactgt
     6301 aattgcaaac tacttacaat atcagtgata tcagattgat gtttttgtcc atagtaagga
     6361 ataattgtaa attcccaagc aggaatcaat ttctttaatg aggcttccaa aattgttgct
     6421 ttttgcgtct tgtatttaaa ctggagtgat ttattgacaa tatcgaaact caacgaattg
     6481 cttatgatag tattatagct catgaatgtg gctctcttga ttgctgttcc gttatgtgta
     6541 atcatccaac ataaataggt tagttcagca gcacataatg ctattttctc acctgaaggt
     6601 ctttcaaacc tttccacaaa ctgacgaaca agcaccttag gtggtgtttt acataatata
     6661 tcaaattgtg gcatgtcgac gattattagt taaaccactg caaaaagttg gggaaaattt
     6721 tgcccatttt tataccgtgt cttcgtctat cgcctccccc actccccaat ctttgaatta
     6781 ttccgaaata ttcagcgaac ggggtgtaca caaaaactaa cattctcaac tgcataattt
     6841 gaaaaatggc gtgggacaag aaaaaaaaaa aattctcaac catagcaatc atggaatacg
     6901 gtaaatttgt gttgttcggt gactccatca cccagtttag ttgtacccag tatggctttc
     6961 atccagcatt acagaatgtg tatatccgaa aattggatgt tattaaccgt ggtttcagtg
     7021 gctacaactc agagcacgct agacaaattc ttccaaaaat tttagagtcg gaaaccaata
     7081 tcaaattgat gacaatattt tttggaacta acgatgcata cgactacatc aatgaaatcc
     7141 agacagtcga gttagacaga tataaagata atttaagtgt aatggtacag atggtactag
     7201 acaaaaatat caaaccaatc attattggat ccgaagttcc tattctctag aaagtatagg
     7261 aacttcctcg agctatttat acaattcatc catacccata acatcagtga atgctttttg
     7321 ccattctttg aaattcaatt cagttttaga atgttttaat tcagtttttc taaaaacata
     7381 cattggttga tttttcaaat aattagcagc cattggttta gcgaaagtat aagtagttct
     7441 agcagttgat ctatatcttt taccattacc agtagtataa gaccatttaa aagttgaaat
     7501 aatagtttta tcatttggat aagttttctt tgatctacac caatcagcag cagttaatga
     7561 attagtcata actggaccat cagctggaaa accagtacct ttaacttgag cttcaccttt
     7621 aatatgagaa ccttcataag tatatctata attaacagtc aatgaagcac catcttcgaa
     7681 ttgcatagtt ctatgaactt gataaccaga accatcaacc atagcagctt gaaatggtga
     7741 cattccatct ggatatggca aatattgatg gaaaccataa ccaatatgtg gaaccaaaat
     7801 ccatggagaa aattgcaaat ctcctttagt tgatttcaaa ttcaattctt cataaccatc
     7861 atttggatta ccagtacctt gaccaaccat atcaaaatca acaccattaa tactaccaaa
     7921 aatatgtaat tcatgtgttg caggcaaact agccatatta tcttcttcac ctttagaaac
     7981 cataccacta ccaccctcga gggggggccc ggtacccaat tcgccctata gtgagtcgta
     8041 ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     8101 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     8161 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga aattgtaagc
     8221 gttaatattt tgttaaaat
//
