LOCUS pRB895 8239 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION C-terminal fusions of C. albicans optimized mNeonGreen fluorescent
protein..
ACCESSION .
VERSION .
KEYWORDS pRB895
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 May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8239
/organism="synthetic DNA construct"
/mol_type="other DNA"
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 2941..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(8210..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
//
