LOCUS pRB897 8224 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION C-terminal fusions of C. albicans optimized mScarlet fluorescent
protein..
ACCESSION .
VERSION .
KEYWORDS pRB897
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8224)
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 8224)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8224
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 372..1643
/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(3900..3933)
/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(3935..3951)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3935..3951)
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(3988..4008)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(3988..4006)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(4027..4043)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4027..4043)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4040..4062)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4051..4067
/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(4075..4105)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4120..4141
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4258..4275)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4429..5017)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4509..4528)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 5237..5339
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS 5340..5999
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(5406..5425)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(6488..6592)
/gene="bla"
/label=AmpR promoter
rep_origin 6619..7075
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6706..6725)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6916..6937
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 7201..7223
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 7215..7232
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 7216..7232
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 7242..7261
/label=T7
/note="T7 promoter, forward primer"
promoter 7242..7260
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind complement(8004..8037)
/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)"
ORIGIN
1 tgtctagcgt gctctgagtt gtagccactg aaaccacggt taataacatc caattttcgg
61 atatacacat tctgtaatgc tggatgaaag ccatactggg tacaactaaa ctgggtgatg
121 gagtcaccga acaacacaaa tttaccgtat tccatgattg ctatggttga gaattttttt
181 tttttcttgt cccacgccat ttttcaaatt atgcagttga gaatgttagt ttttgtgtac
241 accccgttcg ctgaatattt cggaataatt caaagattgg ggagtggggg aggcgataga
301 cgaagacacg gtataaaaat gggcaaaatt ttccccaact ttttgcagtg gtttaactaa
361 taatcgtcga catgccacaa tttgatatat tatgtaaaac accacctaag gtgcttgttc
421 gtcagtttgt ggaaaggttt gaaagacctt caggtgagaa aatagcatta tgtgctgctg
481 aactaaccta tttatgttgg atgattacac ataacggaac agcaatcaag agagccacat
541 tcatgagcta taatactatc ataagcaatt cgttgagttt cgatattgtc aataaatcac
601 tccagtttaa atacaagacg caaaaagcaa caattttgga agcctcatta aagaaattga
661 ttcctgcttg ggaatttaca attattcctt actatggaca aaaacatcaa tctgatatca
721 ctgatattgt aagtagtttg caattacagt tcgaatcatc ggaagaagca gataagggaa
781 atagccacag taaaaaaatg cttaaagcac ttctaagtga gggtgaaagc atctgggaga
841 tcactgagaa aatactaaat tcgtttgagt atacttcgag atttacaaaa acaaaaactt
901 tataccaatt cctcttccta gctactttca tcaattgtgg aagattcagc gatattaaga
961 acgttgatcc gaaatcattt aaattagtcc aaaataagta tttgggagta ataatccagt
1021 gtttagtgac agagacaaag acaagcgtta gtaggcacat atacttcttt agcgcaaggg
1081 gtaggatcga tccacttgta tatttggatg aatttttgag gaattctgaa ccagtcctaa
1141 aacgagtaaa taggaccggc aattcttcaa gcaataaaca ggaataccaa ttattaaaag
1201 ataacttagt cagatcgtac aataaagctt tgaagaaaaa tgcgccttat tcaatctttg
1261 ctataaaaaa tggcccaaaa tctcacattg gaagacattt gatgacctca tttctttcaa
1321 tgaagggcct aacggagttg actaatgttg tgggaaattg gagcgataag cgtgcttctg
1381 ccgtggccag gacaacgtat actcatcaga taacagcaat acctgatcac tacttcgcac
1441 tagtttctcg gtactatgca tatgatccaa tatcaaagga aatgatagca ttgaaggatg
1501 agactaatcc aattgaggag tggcagcata tagaacagct aaagggtagt gctgaaggaa
1561 gcatacgata ccccgcatgg aatgggataa tatcacagga ggtactagac tacctttcat
1621 cctacataaa tagacgcata taagagtgaa attctggaaa tctggaaatc tggttttgta
1681 ttcttgttat tcttcttttt gttattacat atataacttg ttactttttt aaaaaaatct
1741 ttgtatattt tataaatata taaaactaaa tttaagaaaa agagaaaaat gttttatttg
1801 agagattgaa attttacttg aatttagctt agcttttata aagtattatt atgtaaaaaa
1861 acaaaacaaa tatacattaa aaagttaaga ctataaaata gccacccaag gcatttctat
1921 atcttgttgt tgttgttttc atcttctgta tcagaggaac ttattttatt attttcgtca
1981 cgggtatttt ctcttgtttg atgattcatc ccattcattc catcataaaa tgtcgagcgt
2041 caaaactaga gaataataaa gaaaacgatc ttttcaaaaa gaaaaaacct tttagttttc
2101 ctttgttgtt gttgtgggtg tgtgctattt atattatata gtttactcat aataccataa
2161 aatattcggt ttgattaggt tattttaata agctaatttg tttctaatcg tgtaatttat
2221 gctgtgtata ttaagtagtg tgtgcactgc ccaaaaatgt ttgttgttta tagtcggtta
2281 aagagaaaaa agaaaaaaag atccatacac acacgttaat tagttgttca acgtaataca
2341 ctcatatttt gttcttattt gctttcggtc gctgttctca ccaagattta ttgccaacga
2401 aacaattttt ttttatatat tttcagattt ttcttttttt cctttccttt ccttttctaa
2461 ttttcactcc tggttttctt tctttcttag aaacattatc tcgatattaa tattaaaaaa
2521 atataatcat tcaaaatgga cggtggtatg ttttagttta gcttcaattc taattgattg
2581 attaatcagt tgattggttt caatatgaca aatgggtagg gtgggaaaac ttcattttca
2641 attcagatca aacttttttg ttgtcgacat aatatttctc gtttgggatg ttactgtcac
2701 attaataata cacacacatc agcttataat tttgaaagta atttatcaga tatgttgtga
2761 cgatcagtgg aaatggctaa cttcaatgta tctgttcttc ccctttttca aagttcacgt
2821 tttttgattg attgattgat ctgtcggcag tggtttcaaa accattcggt gagtaatcct
2881 atcaatcaat gttacgacaa aaggctcaat attcaaaatt gcaatgtttt atgttttcct
2941 acgtgtactt gtgcaaggca attgattcaa cattgctttt ggtgtttgac gagtttctag
3001 tttggacttg tgttgttatc tgggctatac agatttcccg gctcactatg aatttttttt
3061 ttcgacgctc agtgcacaca actataaaca acacaaacac aaacacagca agaaaaaaaa
3121 aaaacgaaca ttgaattgaa accaagccaa ctgaaaaatt ccttatttaa atgactgtca
3181 tactaaccca tttttataga agaagttgct gctttagtta tcgataacgg ttctcatatg
3241 aaaatttcgg tgatccctga gcaggtggcg gaaacattgg atgctgagaa ccatttcatt
3301 gttcgtgaag tgttcgatgt gcacctatcc gaccaaggct ttgaactatc taccagaagt
3361 gtgagcccct accggaagga ttacatctcg gatgatgact ctgatgaaga ctctgcttgc
3421 tatggcgcat tcatcgacca agagcttgtc gggaagattg aactcaactc aacatggaac
3481 gatctagcct ctatcgaaca cattgttgtg tcgcacacgc accgaggcaa aggagtcgcg
3541 cacagtctca tcgaatttgc gaaaaagtgg gcactaagca gacagctcct tggcatacga
3601 ttagagacac aaacgaacaa tgtacctgcc tgcaatttgt acgcaaaatg tggctttact
3661 ctcggcggca ttgacctctt cacgtataaa actagacctc aagtctcgaa cgaaacagcg
3721 atgtactggt actggttctc gggagcacag gatgacgcct aacatatgtg aagtgtgaag
3781 ggggagattt tcactttatt agatttgtat atatgtataa taaataaata aataagttaa
3841 ataaataatt agataagggt ggtaattatt actatttaca atcaaaggtg gtcctgcagg
3901 aagttcctat actttctaga gaataggaac ttcagatcca ctagttctag agcggccgcc
3961 accgcggtgg agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
4021 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
4081 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
4141 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
4201 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
4261 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
4321 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
4381 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
4441 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
4501 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
4561 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
4621 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
4681 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
4741 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
4801 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
4861 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
4921 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
4981 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
5041 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
5101 aaaaaggatc ttcacctaga tccttttcga ccgaataaat acctgtgacg gaagatcact
5161 tcgcagaata aataaatcct ggtgtccctg ttgataccgg gaagccctgg gccaactttt
5221 ggcgaaaatg agacgttgat cggcacgtaa gaggttccaa ctttcaccat aatgaaataa
5281 gatcactacc gggcgtattt tttgagttgt cgagattttc aggagctaag gaagctaaaa
5341 tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac
5401 attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata
5461 ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc
5521 acattcttgc ccgcctgatg aatgctcatc cggaattacg tatggcaatg aaagacggtg
5581 agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa
5641 cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt
5701 cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga
5761 atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg
5821 ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg
5881 acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg
5941 tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat
6001 ttttttaagg cagttattgg tgcccttaaa cgcctggttg ctacgcctga ataagtgata
6061 ataagcggat gaatggcaga aattcgaaag caaattcgac ccggtcgtcg gttcagggca
6121 gggtcgttaa atagccgctt atgtctattg ctggtttacc ggtttattga ctaccggaag
6181 cagtgtgacc gtgtgcttct caaatgcctg aggccagttt gctcaggctc tccccgtgga
6241 ggtaataatt gacgatatga tccttttttt ctgatcaaaa agtgctcatc attggaaaac
6301 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
6361 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
6421 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
6481 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
6541 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
6601 cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
6661 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
6721 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
6781 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
6841 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
6901 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
6961 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaaatgag
7021 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttccatt
7081 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
7141 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
7201 cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg
7261 gcgaattggg taccgggccc cccctcgagg gtggtagtgg tatggtttct aaaggtgaag
7321 cagttattaa agaattcatg agattcaaag ttcatatgga aggtagtatg aatggtcatg
7381 aatttgaaat tgaaggtgaa ggtgaaggta gaccatatga aggtactcaa actgctaaat
7441 tgaaagttac taaaggtggt ccattaccat tttcatggga tattttgtca ccacaattca
7501 tgtatggttc aagagctttt attaaacatc cagctgatat tccagattat tataaacaat
7561 cttttccaga aggttttaaa tgggaaagag ttatgaattt cgaagatggt ggtgctgtta
7621 ctgttactca agatacttca ttagaagatg gtactttgat ctataaagtt aaattgagag
7681 gtactaattt tccaccagat ggtccagtta tgcaaaagaa aactatgggt tgggaagctt
7741 ctactgaaag attatatcca gaagatggtg ttttgaaagg agatattaaa atggctttga
7801 gattgaaaga tggtggtaga tatttggctg atttcaaaac tacttataaa gctaagaaac
7861 cagttcaaat gccaggtgct tataatgttg atagaaaatt ggatattact tcacataatg
7921 aagattatac tgttgttgaa caatatgaaa gaagtgaagg tagacatagt acaggtggta
7981 tggatgaatt atattagctc gaggaagttc ctatactttc tagagaatag gaacttcgga
8041 tccaataatg attggtttga tatttttgtc tagtaccatc tgtaccatta cacttaaatt
8101 atctttatat ctgtctaact cgactgtctg gatttcattg atgtagtcgt atgcatcgtt
8161 agttccaaaa aatattgtca tcaatttgat attggtttcc gactctaaaa tttttggaag
8221 aatt
//
