LOCUS       sequence-292547-        8424 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluoroescent marker for use in Cryptococcus neoformans. Contains a 
            codon optimised mMaroon1 marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Fungal 
            Genet Biol."; date: "2020-05"; volume: "138:103365. doi"; pages: " 
            10.1016/j.fgb.2020.103365. Epub 2020 Mar 4"
COMMENT     Sequence Label: pGWKS6
FEATURES             Location/Qualifiers
     source          1..8424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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(join(5518..5524,5525..5542,5543..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(join(6631..7422,7423..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
//
