LOCUS Exported 8424 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Fluoroescent marker for use in Cryptococcus neoformans. Contains a codon optimised mMaroon1 marker.. ACCESSION . VERSION . KEYWORDS pGWKS6 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8424) AUTHORS Spencer GWK, Chua SMH, Erpf PE, Wizrah MSI, Dyba TG, Condon ND, Fraser JA TITLE Broadening the spectrum of fluorescent protein tools for use in the encapsulated human fungal pathogen Cryptococcus neoformans. JOURNAL Fungal Genet Biol. 2020 May;138:103365. doi: 10.1016/j.fgb.2020.103365. Epub 2020 Mar 4. PUBMED 32145317 REFERENCE 2 (bases 1 to 8424) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8424 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(1025..1588) /codon_start=1 /gene="Streptomyces noursei nat1" /product="nourseothricin acetyltransferase" /label=NrsR /note="confers resistance to nourseothricin" /translation="TLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGFTLR EVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNRRLT VEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFTLCG LDTALYDGTASDGEQALYMSMPCP" primer_bind 2696..2718 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2710..2727 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2711..2727 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 2737..2756 /label=T7 /note="T7 promoter, forward primer" promoter 2737..2755 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5431..5451) /label=T3 /note="T3 promoter, forward primer" promoter complement(5431..5449) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(5470..5486) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5470..5486) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5483..5505) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5494..5510 /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(5518..5548) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5563..5584 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5701..5718) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5872..6460) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5952..5971) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6631..7491) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 7254..7273 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7492..7596) /gene="bla" /label=AmpR promoter ORIGIN 1 accaaaccag tctgtgccaa gtggattgct tatgcatcga gtggcgagcc cactgacctc 61 tgggcccctt ttttgaacta taaaaagcgg ccaagtctga tttcggtctc acaatcatca 121 acatcgactt ctcttttcgc ctcgatcaac ctaaacaacg gtcactcagc catgaactgg 181 acaatggaat atttcgcctt gccttggatc tttggcctct agactcaatc aacgtcagga 241 taaaggtgag cataggttgc tcactcactg tagccttttt ttcccttctt cttggtccag 301 atggttccca aggctctata aagtgtatct gtcagttcca cttccttact ccgaagcgat 361 tcagagttgt aaagtgcgtc tgaccccctg attatatctc aaagtccctt cacaaattca 421 gacatgatgc gataaagtgc cggattgtca ttcgcgtctt gtgactgtat agaaagtgcc 481 ggcatcctct tttcttttcc tctttggccg agcgactcag agttgtgaag cctgctggta 541 tctgtctgtt cccttgccct gataaagaat atcgaagagc atttctttga ttcacaatac 601 gttgacgaat ggtgtgatgg atcctaagca gcgccagccc tttgaacttg tgtcgaaaaa 661 aaaaaaaaaa cagcatatga tacgtaaagg gaagaaaagg atggagctga ctgttttgac 721 aggcataatt ttgagcgtgc ttcattggcc ggtttatctg tattaacacg gaagagatgt 781 agaaactagc ttcctggttt cagagacaag gagccatgaa gatcctgagg attcaacggc 841 gttgctgagg gtgggaagga tggtattgtt gtctgtagat agattaatgg gcatataatg 901 tcaaaaaaga tgattgacac gatgtctgtc tcgaattcgc agtgtgctaa aattgtatca 961 cttctctaat atgtatatat acaccctcta aggaaaatta accccttacc gccttcacga 1021 attcttaggg gcagggcatg ctcatgtaga gcgcctgctc gccgtccgag gcggtgccgt 1081 cgtacagggc ggtgtccagg ccgcagaggg tgaaccccat ccgccggtac gcgtggatag 1141 ccggtgcgtt aacgttggtg acctccagcc agaggtgccc ggcgccccgc tcgcgggcga 1201 actccgtcgc gagccccatc aacgcgcgcc cgaccccgtg cccccggtgc tccggggcga 1261 cctcgatgtc ctcgacggtc agccggcggt tccagccgga gtacgagacg accacgaagc 1321 ccgccaggtc gccgtcgtcc ccgtacgcga cgaacgtccg ggagtccggg tcgccgtcct 1381 ccccgtcgtc cgattcgtcg tccgattcgt cgtcggggaa caccttggtc aggggcgggt 1441 ccaccggcac ctcccgcagg gtgaagccgt ccccggtggc ggtgacgcgg aagacggtgt 1501 cggtggtgaa ggacccatcc agtgcctcga tggcctcggc gtcccccggg acactggtgc 1561 ggtaccggta agccgtgtcg tcaagagtgg cggccgccat agacatgttg ggcgagttta 1621 ctaatggaaa aagacaatta gcaaagaggc tgcatattag gagacgcggt gaagattggg 1681 gaggacaata gggtccgtgc accaaaaagt cctgacggca gctctgttca acgcacatcc 1741 atctcttcca gctatgcgtg tggcaccagc acggatggat tgttgcacca aaaggacaag 1801 gcgagatgga tgcaggtcga ggcgatgcgg tgctatcggc cctcgttgtc gaacgccacg 1861 tcgtcctctc ggcgaccgtc aatcctggaa ggaaatgacc aagcgtgcac ccagcatcca 1921 gcatccaaca tccagcaccc agcacctagc acccatgatc caatggccgg catgcagatc 1981 agcgtgggga gagcaccatc tagcagcccc tggcaggccc agcattgtgc tcggatgtgc 2041 gataggcggt gggtgcgata aggtggatta gtggaggatg gaggactgag tggcgggcaa 2101 gggaaggggg cgggaagatg actcaccttg ggcagtgggt gggggatgcg gtatagggga 2161 tgtatgtgga tgctggcgga ggatagaagc tgtaagctga gctggtgcca acggtgacgc 2221 tgtgagagtg gtctcggcga ccctgacggc gactcggcgt taccccgtgt ggcacaccgg 2281 acgcgcgtcg agtgacgttg ctgctattgt tgttaccatc atcctctcct ccgacaacca 2341 tacactcagt gcacaccatg ccactcgaat cctgcatgct tatgtgagtc ctccccttcc 2401 cgtgcgccgc tctccagctc acatcctcgc agccatggac tcaaccctat ctcggtctat 2461 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa atgagctgat 2521 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt ccattcgcca 2581 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag 2641 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag 2701 tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa 2761 ttgggtaccg ggccccccct cgagcattct atgctatacg gtacaagatt gtggctacta 2821 tgcaagctcc atagcgaatg atcttacact gaccttgtcg accgttctga atgattaaca 2881 atggtcgcaa gtgtggttga cccacctgcg ggcgcttctt aactatctat catcggtatg 2941 tcaagtatca ctccctttaa acaaaagaat aagagcataa tacagcagtc tcatcatggc 3001 aacccttcgc gcaatgtaca aaaagcagcc agaagccccc acgcacaagc aataatgtcg 3061 tgttctaccc tgccatgaac aagcgtcatg gccattagcg aacgccccaa tattcagccc 3121 cattcgccaa gcaagttgcc tcgccccctt cccagaagat catgggcatc tcgtcttctt 3181 ctaatttcgc attttgtgaa gctccattca tgcacgtcgg cttctggacg actcttatca 3241 tatgcattgg cctgctcggt gtgaggatga gccactccca catgtttaga cggcaggggc 3301 ctcttgccac ccgttgaatt tttggtgcga agacgtgcct gggaacgacc tccactttaa 3361 atttctccgt tgttgcggtc gctccatcct cctcacattt cttgcctcct aagtcccatg 3421 tgccacactc ggtgaaaaac ggagtttgat acggatgaaa tttccagtga gcttttcaca 3481 ttctggaggt ttcgaaaaaa attgtgggat tttctgaatt aatccgagaa tccgccctgc 3541 ccatcaaccg gaaagaatta aatccctcgc acattccttt ctcccaccac cttctttatc 3601 tctacattct ttaaccaaaa cacctaggtg agtacccaaa gtcttttgac acctttgaaa 3661 atccgcgcca gtcgtagggc acgtaaaaca attctcgctg gtggcttctg caggtgaagt 3721 cggggcagag tggagctgag catggtcgac cacccacatc attctgtctt gtttttggcg 3781 atgtcttcca tgcttttgcc ctcatctgtt gcgtcgactg cagagtactg ttttgccact 3841 actgagctca acatgggctt gtgtgtggtg aagaaaaaaa tttggtggct cttgaaaatt 3901 ttcaacccat cggatacatg cccttccaac tcacaacgcc ctaacacttt attgtcctat 3961 cattgcctct tcgcacatgc taacgatctc cacagaaaac ttcaaaatgg tctccaaggg 4021 cgaggagctc atcaaggaga acatgcacac caagctctac ctcactggca ccgtcaacaa 4081 ccactacttc gagtgcaccg ccgagggcga gggcaagccc tacgagggca cccagaccaa 4141 caggatcaag gtcgtccgtg gtggccccct ccccttcgct ttcgacatcc tcgccccctg 4201 cttcatgtac ggctccaaga ccttcattaa ccaccctcct gacattcccg actacttcaa 4261 gcagtccttc cctgagggct tcacttggga gaggaccacc gtttacgagg acggcggcac 4321 tctcactgct actcaggaca cctccctcca ggacggctgc ctcatctaca acgtccaggt 4381 caggggcgag aacttccctt ccaacggccc tgtcatgcag aagaagactc tcggctggga 4441 ggcctccacc gagaccctct accccgctga cggctccctc gagggcaggc tcgactgggc 4501 cctcaagctc gtcggcggcg gccacctcca ctgcaggctc gagaccactt acaggtccaa 4561 gaagcccgct aagaacctca agatgcccgg cgtctacttc attgacagga ggctcgagag 4621 gatcaaggag gccgacaacg agacctacgt cgagcagcac gaggtcgctg tcgccaggta 4681 ctgcgacctc ccttccaagc tcggccacaa gcttaacggc atggacgagc tctacaagta 4741 gaaggttgag cgcgtaatag tgggttggtc ttgtctctat ccattcatct gtgagacata 4801 ctgatttcaa ctttttcatc agataataag gatggtgtga gataagcaag gctgggttat 4861 gaggtttggg actagatggt gtttcgcgtc taaaggtgag tcttcgttgg ttcaatcaca 4921 ttgatctgat cgctgatcaa ttgtcagcct ctgaatgtgc tcttgaagaa aatggttgtc 4981 tgtaagaatc cagaaccaaa aacggtcatt tcaagtatgt atcctccgtg aagttccttt 5041 agctcttgct aatattggac tttagcgtgc tttagccaaa gacagggtcg cgaggttaag 5101 aggatgggcc aggaaaaatc taagttagat ttgtcacaca aagaaaaaga ttgtctcagt 5161 tttggttttg tttggtttga tttatgcata ggaaatattt atgatcgaaa tcactcattt 5221 cctgcttcgt aaatatcatc tactatatgc ttcgtaaatg tccatctcct acatattgtg 5281 gatatgataa atgcaatcct ctcgtctaca ctctcggctt atgcctttgt cttctttgcg 5341 ctccttttag ccttcttttt attcctagtc ttcaagccca tcactagttc tagagcggcc 5401 gccaccgcgg tggagctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc 5461 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa 5521 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac 5581 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca 5641 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc 5701 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 5761 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 5821 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 5881 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 5941 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 6001 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 6061 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 6121 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 6181 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 6241 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 6301 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 6361 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 6421 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 6481 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 6541 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 6601 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 6661 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac 6721 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg 6781 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag 6841 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 6901 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt 6961 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt 7021 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt 7081 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct 7141 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt 7201 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac 7261 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa 7321 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa 7381 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca 7441 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct 7501 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga 7561 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc 7621 tgacgcgccc tgtagcggca agatctgccc atagacttca aatcatggaa ttggacgttt 7681 atcttcttgc tttggatatc caccctatcg tacaatcttg gcactttgga tcatacgagg 7741 cactttgcgg actagggaat ggttatcatc aatttcttgg acctctccat gacacctggc 7801 atcgtcgagc taccacagcc ccctgaattc ttaaaggcga accccccttc ctaggaacct 7861 tgagcgctgg actggatggc actctgagac cgcctatgct taaggacgcg agcttaccct 7921 tccattgatg gaagtcgatc atttatgata atagaacacg gcggaacttg actttacagg 7981 ccaagggaag caacaggaga agaagctaag gttataacag gctcagcata tttgtatata 8041 cggtgccatt caacgctgaa cactttgtac tcacggtctg cccgttgaat aacaccatct 8101 gctgatgatc atcccaattt gtgagtctta tttgccattg atcctggtgt ataaccgata 8161 ctgaagccct ttaggtctga tcagttatag acgtggtata ttcattcttc ctagattagc 8221 tatgccgaaa gagatggata ccttgatgtg actggaatgc catgaagcag gagattgtag 8281 cagctcaaat tggagaagga tgttgcaacc gtcggccgtg aaggtcttgt catatctttg 8341 tctgcctggg agaatatgta caagtgcccc gatgttactg atggaaaaca tcgtgtggcg 8401 cgccacgata cttgtgttaa ttaa //