LOCUS       sequence-443459-       10030 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GAL7(p)-amdS:mRuby3 fusion reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10030)
  AUTHORS   Phillips-Rose LS, Yu CK, West NP, Fraser JA
  TITLE     A Chimeric ORF Fusion Phenotypic Reporter for Cryptococcus 
            neoformans.
  JOURNAL   J Fungi (Basel). 2024 Aug 12;10(8):567. doi: 10.3390/jof10080567.
  PUBMED    39194893
REFERENCE   2  (bases 1 to 10030)
  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: "J Fungi 
            (Basel)."; date: "2024-08-12"; volume: "10(8):567. doi"; pages: " 
            10.3390/jof10080567"
COMMENT     Sequence Label: pLSPR26
FEATURES             Location/Qualifiers
     source          1..10030
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1823..2623)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MSMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1934..1953)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2544..2563
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3713..3735
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3727..3744
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3728..3744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3754..3773
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3754..3772
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7841..7861)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7841..7859)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7880..7896)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7880..7896)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7893..7915)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7904..7920
                     /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(7928..7934,7935..7952,7953..7958))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7973..7994
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8111..8128)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8282..8870)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8362..8381)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9041..9832,9833..9901))
                     /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     9664..9683
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9902..10006)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgacgcgccc tgtagcggca agatctgccc atagacttca aatcatggaa ttggacgttt
       61 atcttcttgc tttggatatc caccctatcg tacaatcttg gcactttgga tcatacgagg
      121 cactttgcgg actagggaat ggttatcatc aatttcttgg acctctccat gacacctggc
      181 atcgtcgagc taccacagcc ccctgaattc ttaaaggcga accccccttc ctaggaacct
      241 tgagcgctgg actggatggc actctgagac cgcctatgct taaggacgcg agcttaccct
      301 tccattgatg gaagtcgatc atttatgata atagaacacg gcggaacttg actttacagg
      361 ccaagggaag caacaggaga agaagctaag gttataacag gctcagcata tttgtatata
      421 cggtgccatt caacgctgaa cactttgtac tcacggtctg cccgttgaat aacaccatct
      481 gctgatgatc atcccaattt gtgagtctta tttgccattg atcctggtgt ataaccgata
      541 ctgaagccct ttaggtctga tcagttatag acgtggtata ttcattcttc ctagattagc
      601 tatgccgaaa gagatggata ccttgatgtg actggaatgc catgaagcag gagattgtag
      661 cagctcaaat tggagaagga tgttgcaacc gtcggccgtg aaggtcttgt catatctttg
      721 tctgcctggg agaatatgta caagtgcccc gatgttactg atggaaaaca tcgtgtggcg
      781 cgccacgata cttgtgttaa ttaaaccaaa ccagtctgtg ccaagtggat tgcttatgca
      841 tcgagtggcg agcccactga cctctgggcc ccttttttga actataaaaa gcggccaagt
      901 ctgatttcgg tctcacaatc atcaacatcg acttctcttt tcgcctcgat caacctaaac
      961 aacggtcact cagccatgaa ctggacaatg gaatatttcg ccttgccttg gatctttggc
     1021 ctctagactc aatcaacgtc aggataaagg tgagcatagg ttgctcactc actgtagcct
     1081 ttttttccct tcttcttggt ccagatggtt cccaaggctc tataaagtgt atctgtcagt
     1141 tccacttcct tactccgaag cgattcagag ttgtaaagtg cgtctgaccc cctgattata
     1201 tctcaaagtc ccttcacaaa ttcagacatg atgcgataaa gtgccggatt gtcattcgcg
     1261 tcttgtgact gtatagaaag tgccggcatc ctcttttctt ttcctctttg gccgagcgac
     1321 tcagagttgt gaagcctgct ggtatctgtc tgttcccttg ccctgataaa gaatatcgaa
     1381 gagcatttct ttgattcaca atacgttgac gaatggtgtg atggatccta agcagcgcca
     1441 gccctttgaa cttgtgtcga aaaaaaaaaa aaaacagcat atgatacgta aagggaagaa
     1501 aaggatggag ctgactgttt tgacaggcat aattttgagc gtgcttcatt ggccggttta
     1561 tctgtattaa cacggaagag atgtagaaac tagcttcctg gtttcagaga caaggagcca
     1621 tgaagatcct gaggattcaa cggcgttgct gagggtggga aggatggtat tgttgtctgt
     1681 agatagatta atgggcatat aatgtcaaaa aagatgattg acacgatgtc tgtctcgaat
     1741 tcgcagtgtg ctaaaattgt atcacttctc taatatgtat atatacaccc tctaaggaaa
     1801 attaacccct taccgccttc actcagaaga actcgtcaag aaggcgatag aaggcgatgc
     1861 gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc
     1921 caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac
     1981 ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca
     2041 agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgcgc gccttgagcc
     2101 tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga
     2161 caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga
     2221 atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata
     2281 ctttctcggc aggagcaagg tgggatgaca ggagatcctg ccccggcact tcgcccaata
     2341 gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg
     2401 tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca
     2461 ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat
     2521 cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg
     2581 ccggagaacc tgcgtgcaat ccatcttgtt caatcataga catgttgggc gagtttacta
     2641 atggaaaaag acaattagca aagaggctgc atattaggag acgcggtgaa gattggggag
     2701 gacaataggg tccgtgcacc aaaaagtcct gacggcagct ctgttcaacg cacatccatc
     2761 tcttccagct atgcgtgtgg caccagcacg gatggattgt tgcaccaaaa ggacaaggcg
     2821 agatggatgc aggtcgaggc gatgcggtgc tatcggccct cgttgtcgaa cgccacgtcg
     2881 tcctctcggc gaccgtcaat cctggaagga aatgaccaag cgtgcaccca gcatccagca
     2941 tccaacatcc agcacccagc acctagcacc catgatccaa tggccggcat gcagatcagc
     3001 gtggggagag caccatctag cagcccctgg caggcccagc attgtgctcg gatgtgcgat
     3061 aggcggtggg tgcgataagg tggattagtg gaggatggag gactgagtgg cgggcaaggg
     3121 aagggggcgg gaagatgact caccttgggc agtgggtggg ggatgcggta taggggatgt
     3181 atgtggatgc tggcggagga tagaagctgt aagctgagct ggtgccaacg gtgacgctgt
     3241 gagagtggtc tcggcgaccc tgacggcgac tcggcgttac cccgtgtggc acaccggacg
     3301 cgcgtcgagt gacgttgctg ctattgttgt taccatcatc ctctcctccg acaaccatac
     3361 actcagtgca caccatgcca ctcgaatcct gcatgcttat gtgagtcctc cccttcccgt
     3421 gcgccgctct ccagctcaca tcctcgcagc catggactca accctatctc ggtctattct
     3481 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaaatg agctgattta
     3541 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcca ttcgccattc
     3601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     3661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     3721 cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg
     3781 ggtaccgggc cccccctcga tagtgaagtc gtattgtctc tgcaatgtaa ttctctttgc
     3841 catgcatatg cttgagcctg tgccttctct ttaatcgaca gcactgacaa tctcttgcag
     3901 tttcgtcgca gtagtcacgc gtatatgttt ccatagttcc ggcgggaagt tatcttacta
     3961 aatacggtat cgaaggaatt acacaatatt tgtactccgg cggtggaaac aggtcttgtc
     4021 gaaccctgat gaagcgaaag ctaagaggtt cgaaagtagc aaaatcatac tttaaaatct
     4081 tttatacgcg agtccaactt cgctgacaag aaaatggtag tgggccttgc gaatgaatga
     4141 ggagagaaag ttttagctgt ggattcccag caggcctcct tacttaagtt tgattaaaaa
     4201 ttattacgaa aagatatata tagtaataaa tttgaaatga actaatcata atcgaaaaat
     4261 tcccgttcaa cataaattcc cgttcattta aattcccgtt caacataaat tcccgttcat
     4321 ttaaattccc gttcaacata aaattcccgt tcataaattc ccatggcatt catcatcatg
     4381 aatcctttgt tgtatagtta ttatataacg gatattcccg ttcgtatcca ttgcatatct
     4441 tatcgcatag tcttcccgtt catcgtctca tcatcaagtt attgattgtc aattcacgct
     4501 tacatattcc cgttcttctg tctaccatct atctatcaca tcatctgtga attcttcccg
     4561 tttttatggg aagactgtcg tcatgttgtc aattgagcag cagacataat cgatgctata
     4621 tatctagagc ttcacggctc aggctgaact tggattaatg tggactactt tcacaacgaa
     4681 tacttatcag cactcaattc tctcctgaga atgcctcagt cctgggagga gctcgccgct
     4741 gacaagcgtg cccgtctcgc taagaccatc cctgacgagt ggaaggtcca gaccctccct
     4801 gccgaggact ccgttattga cttccctaag aagtctggca tcctttctga ggccgagctc
     4861 aagatcactg aggcctccgc tgctgacctt gtctccaagc tcgccgccgg tgagttgacc
     4921 tctgtcgagg ttacccttgc tttctgtaag cgagctgcta tcgcccagca gttgactaac
     4981 tgcgcccacg agttcttccc tgacgccgct ctcgcccagg ctagggagct cgacgagtac
     5041 tacgctaagc acaagaggcc cgttggtcct ctccacggcc tccccatctc tctcaaggac
     5101 cagcttcgag tcaagggcta cgagacttct atgggctaca tctcttggct taacaagtac
     5161 gacgagggcg actctgttct cactaccatg ctccgtaagg ccggtgccgt cttctacgtc
     5221 aagacctctg tcccccagac cctcatggtc tgcgagactg tcaacaacat catcggccgt
     5281 accgtcaacc ctcgtaacaa gaactggtct tgcggcggct cctctggtgg tgagggtgcc
     5341 atcgttggca ttcgtggtgg cgtcatcggt gttggtaccg acattggtgg ctctattcga
     5401 gtccccgccg ccttcaactt cctctacggt cttaggccct cccacggccg actcccctac
     5461 gctaagatgg ccaactccat ggagggtcag gagaccgtcc actccgttgt cggccccatt
     5521 acccactctg ttgaggacct ccgtctcttc accaagtccg tcctcggtca ggagccttgg
     5581 aagtacgact ccaaggtcat ccccatgccc tggcgtcagt ccgagtctga cattattgcc
     5641 tccaagatca agaacggcgg cctcaacatc ggctactaca acttcgacgg caacgtcctt
     5701 cctcaccctc ctatcctccg tggcgtcgag accaccgtcg ccgctctcgc caaggccggt
     5761 cacaccgtca ccccctggac cccttacaag cacgacttcg gccacgacct catctcccac
     5821 atctacgccg ctgacggctc cgccgacgtt atgcgtgaca tctccgcttc cggcgagccc
     5881 gccattccta acatcaagga ccttctcaac cccaacatca aggctgttaa catgaacgag
     5941 ctctgggaca cccacctcca gaagtggaac taccagatgg agtaccttga gaagtggcga
     6001 gaggctgagg agaaggccgg caaggaggtg agtagatcac aatttgtaac aggttggatg
     6061 ataagtcccc ggtctaacat gtagcgacga cactgaagct cacttcttgt agctcgacgc
     6121 catcatcgcc cccattaccc ctaccgctgc cgttcgacac gaccagttcc gatactacgg
     6181 ctacgcctct gtcatcaacc tcctcgactt cacctccgtc gttgttcccg ttaccttcgc
     6241 cgacaagaac atcgacaaga agaacgagtc cttcaaggcc gtttccgagc ttgacgccct
     6301 cgtccaggag gagtacgacc ccgaggccta ccacggcgct cccgttgctg tccaggttat
     6361 cggtcgaagg ctctccgagg agaggacctt ggccattgct gaggaggtcg gcaagttgct
     6421 cggtaacgtc gtcactcctg gttctggcgg tggtggctcc ggcatggtct ctaagggcga
     6481 ggagctcatc aaggagaaca tgcgtatgaa ggtcgtcatg gagggttctg tcaacggcca
     6541 ccagttcaag tgcactggtg agggtgaggg caggccctac gagggtgtcc agaccatgag
     6601 gatcaaggtc atcgagggtg gtcccctccc tttcgccttc gacattcttg ccacttcttt
     6661 catgtacggc tcccgtacct tcatcaagta ccccgccgac atccctgact tcttcaagca
     6721 gtccttccct gagggtttca cttgggagag ggttaccagg tacgaggacg gtggtgtcgt
     6781 caccgtcacc caggacacct cccttgagga cggcgagctc gtctacaacg tcaaggtcag
     6841 gggcgttaac ttcccctcca acggtcccgt catgcagaag aagaccaagg gttgggagcc
     6901 taacactgag atgatgtacc ctgctgacgg tggtctcagg ggttacactg acatcgctct
     6961 caaggttgac ggtggtggcc acctccactg caacttcgtc actacttaca ggtctaagaa
     7021 gaccgtcggc aacatcaaga tgcccggtgt ccacgccgtt gaccaccgtc tcgagaggat
     7081 cgaggagtcc gacaacgaga cctacgttgt ccagagggag gtcgctgttg ccaagtactc
     7141 caaccttggt ggtggcatgg acgagctcta caagtagaag gttgagcgcg taatagtggg
     7201 ttggtcttgt ctctatccat tcatctgtga gacatactga tttcaacttt ttcatcagat
     7261 aataaggatg gtgtgagata agcaaggctg ggttatgagg tttgggacta gatggtgttt
     7321 cgcgtctaaa ggtgagtctt cgttggttca atcacattga tctgatcgct gatcaattgt
     7381 cagcctctga atgtgctctt gaagaaaatg gttgtctgta agaatccaga accaaaaacg
     7441 gtcatttcaa gtatgtatcc tccgtgaagt tcctttagct cttgctaata ttggacttta
     7501 gcgtgcttta gccaaagaca gggtcgcgag gttaagagga tgggccagga aaaatctaag
     7561 ttagatttgt cacacaaaga aaaagattgt ctcagttttg gttttgtttg gtttgattta
     7621 tgcataggaa atatttatga tcgaaatcac tcatttcctg cttcgtaaat atcatctact
     7681 atatgcttcg taaatgtcca tctcctacat attgtggata tgataaatgc aatcctctcg
     7741 tctacactct cggcttatgc ctttgtcttc tttgcgctcc ttttagcctt ctttttattc
     7801 ctagtcttca agcccatcgg tggagctcca gcttttgttc cctttagtga gggttaattg
     7861 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     7921 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     7981 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     8041 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     8101 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     8161 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     8221 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     8281 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     8341 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     8401 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     8461 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     8521 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     8581 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     8641 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     8701 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     8761 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     8821 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     8881 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     8941 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     9001 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     9061 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     9121 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     9181 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     9241 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     9301 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     9361 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     9421 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     9481 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     9541 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     9601 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     9661 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     9721 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     9781 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     9841 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     9901 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     9961 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
    10021 aagtgccacc
//
