LOCUS       sequence-443454-       10030 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GAL7(p)-mRuby3:amdS 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: pLSPR30
FEATURES             Location/Qualifiers
     source          1..10030
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1824..2624)
                     /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(1935..1954)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2545..2564
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3714..3736
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3728..3745
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3729..3745
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3755..3774
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3755..3773
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7842..7862)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7842..7860)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7881..7897)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7881..7897)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7894..7916)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7905..7921
                     /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(7929..7935,7936..7953,7954..7959))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7974..7995
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8112..8129)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8283..8871)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8363..8382)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9042..9833,9834..9902))
                     /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     9665..9684
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9903..10007)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgcgcc ctgtagcggc aagatctgcc catagacttc aaatcatgga attggacgtt
       61 tatcttcttg ctttggatat ccaccctatc gtacaatctt ggcactttgg atcatacgag
      121 gcactttgcg gactagggaa tggttatcat caatttcttg gacctctcca tgacacctgg
      181 catcgtcgag ctaccacagc cccctgaatt cttaaaggcg aacccccctt cctaggaacc
      241 ttgagcgctg gactggatgg cactctgaga ccgcctatgc ttaaggacgc gagcttaccc
      301 ttccattgat ggaagtcgat catttatgat aatagaacac ggcggaactt gactttacag
      361 gccaagggaa gcaacaggag aagaagctaa ggttataaca ggctcagcat atttgtatat
      421 acggtgccat tcaacgctga acactttgta ctcacggtct gcccgttgaa taacaccatc
      481 tgctgatgat catcccaatt tgtgagtctt atttgccatt gatcctggtg tataaccgat
      541 actgaagccc tttaggtctg atcagttata gacgtggtat attcattctt cctagattag
      601 ctatgccgaa agagatggat accttgatgt gactggaatg ccatgaagca ggagattgta
      661 gcagctcaaa ttggagaagg atgttgcaac cgtcggccgt gaaggtcttg tcatatcttt
      721 gtctgcctgg gagaatatgt acaagtgccc cgatgttact gatggaaaac atcgtgtggc
      781 gcgccacgat acttgtgtta attaaaccaa accagtctgt gccaagtgga ttgcttatgc
      841 atcgagtggc gagcccactg acctctgggc cccttttttg aactataaaa agcggccaag
      901 tctgatttcg gtctcacaat catcaacatc gacttctctt ttcgcctcga tcaacctaaa
      961 caacggtcac tcagccatga actggacaat ggaatatttc gccttgcctt ggatctttgg
     1021 cctctagact caatcaacgt caggataaag gtgagcatag gttgctcact cactgtagcc
     1081 tttttttccc ttcttcttgg tccagatggt tcccaaggct ctataaagtg tatctgtcag
     1141 ttccacttcc ttactccgaa gcgattcaga gttgtaaagt gcgtctgacc ccctgattat
     1201 atctcaaagt cccttcacaa attcagacat gatgcgataa agtgccggat tgtcattcgc
     1261 gtcttgtgac tgtatagaaa gtgccggcat cctcttttct tttcctcttt ggccgagcga
     1321 ctcagagttg tgaagcctgc tggtatctgt ctgttccctt gccctgataa agaatatcga
     1381 agagcatttc tttgattcac aatacgttga cgaatggtgt gatggatcct aagcagcgcc
     1441 agccctttga acttgtgtcg aaaaaaaaaa aaaaacagca tatgatacgt aaagggaaga
     1501 aaaggatgga gctgactgtt ttgacaggca taattttgag cgtgcttcat tggccggttt
     1561 atctgtatta acacggaaga gatgtagaaa ctagcttcct ggtttcagag acaaggagcc
     1621 atgaagatcc tgaggattca acggcgttgc tgagggtggg aaggatggta ttgttgtctg
     1681 tagatagatt aatgggcata taatgtcaaa aaagatgatt gacacgatgt ctgtctcgaa
     1741 ttcgcagtgt gctaaaattg tatcacttct ctaatatgta tatatacacc ctctaaggaa
     1801 aattaacccc ttaccgcctt cactcagaag aactcgtcaa gaaggcgata gaaggcgatg
     1861 cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg
     1921 ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca
     1981 cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc
     2041 aagcaggcat cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg cgccttgagc
     2101 ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg
     2161 acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg
     2221 aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat
     2281 actttctcgg caggagcaag gtgggatgac aggagatcct gccccggcac ttcgcccaat
     2341 agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc
     2401 gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac
     2461 aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca
     2521 tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg
     2581 gccggagaac ctgcgtgcaa tccatcttgt tcaatcatag acatgttggg cgagtttact
     2641 aatggaaaaa gacaattagc aaagaggctg catattagga gacgcggtga agattgggga
     2701 ggacaatagg gtccgtgcac caaaaagtcc tgacggcagc tctgttcaac gcacatccat
     2761 ctcttccagc tatgcgtgtg gcaccagcac ggatggattg ttgcaccaaa aggacaaggc
     2821 gagatggatg caggtcgagg cgatgcggtg ctatcggccc tcgttgtcga acgccacgtc
     2881 gtcctctcgg cgaccgtcaa tcctggaagg aaatgaccaa gcgtgcaccc agcatccagc
     2941 atccaacatc cagcacccag cacctagcac ccatgatcca atggccggca tgcagatcag
     3001 cgtggggaga gcaccatcta gcagcccctg gcaggcccag cattgtgctc ggatgtgcga
     3061 taggcggtgg gtgcgataag gtggattagt ggaggatgga ggactgagtg gcgggcaagg
     3121 gaagggggcg ggaagatgac tcaccttggg cagtgggtgg gggatgcggt ataggggatg
     3181 tatgtggatg ctggcggagg atagaagctg taagctgagc tggtgccaac ggtgacgctg
     3241 tgagagtggt ctcggcgacc ctgacggcga ctcggcgtta ccccgtgtgg cacaccggac
     3301 gcgcgtcgag tgacgttgct gctattgttg ttaccatcat cctctcctcc gacaaccata
     3361 cactcagtgc acaccatgcc actcgaatcc tgcatgctta tgtgagtcct ccccttcccg
     3421 tgcgccgctc tccagctcac atcctcgcag ccatggactc aaccctatct cggtctattc
     3481 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaaat gagctgattt
     3541 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc attcgccatt
     3601 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     3661 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     3721 acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat agggcgaatt
     3781 gggtaccggg ccccccctcg atagtgaagt cgtattgtct ctgcaatgta attctctttg
     3841 ccatgcatat gcttgagcct gtgccttctc tttaatcgac agcactgaca atctcttgca
     3901 gtttcgtcgc agtagtcacg cgtatatgtt tccatagttc cggcgggaag ttatcttact
     3961 aaatacggta tcgaaggaat tacacaatat ttgtactccg gcggtggaaa caggtcttgt
     4021 cgaaccctga tgaagcgaaa gctaagaggt tcgaaagtag caaaatcata ctttaaaatc
     4081 ttttatacgc gagtccaact tcgctgacaa gaaaatggta gtgggccttg cgaatgaatg
     4141 aggagagaaa gttttagctg tggattccca gcaggcctcc ttacttaagt ttgattaaaa
     4201 attattacga aaagatatat atagtaataa atttgaaatg aactaatcat aatcgaaaaa
     4261 ttcccgttca acataaattc ccgttcattt aaattcccgt tcaacataaa ttcccgttca
     4321 tttaaattcc cgttcaacat aaaattcccg ttcataaatt cccatggcat tcatcatcat
     4381 gaatcctttg ttgtatagtt attatataac ggatattccc gttcgtatcc attgcatatc
     4441 ttatcgcata gtcttcccgt tcatcgtctc atcatcaagt tattgattgt caattcacgc
     4501 ttacatattc ccgttcttct gtctaccatc tatctatcac atcatctgtg aattcttccc
     4561 gtttttatgg gaagactgtc gtcatgttgt caattgagca gcagacataa tcgatgctat
     4621 atatctagag cttcacggct caggctgaac ttggattaat gtggactact ttcacaacga
     4681 atacttatca gcactcaatt ctctcctgag aatggtctct aagggcgagg agctcatcaa
     4741 ggagaacatg cgtatgaagg tcgtcatgga gggttctgtc aacggccacc agttcaagtg
     4801 cactggtgag ggtgagggca ggccctacga gggtgtccag accatgagga tcaaggtcat
     4861 cgagggtggt cccctccctt tcgccttcga cattcttgcc acttctttca tgtacggctc
     4921 ccgtaccttc atcaagtacc ccgccgacat ccctgacttc ttcaagcagt ccttccctga
     4981 gggtttcact tgggagaggg ttaccaggta cgaggacggt ggtgtcgtca ccgtcaccca
     5041 ggacacctcc cttgaggacg gcgagctcgt ctacaacgtc aaggtcaggg gcgttaactt
     5101 cccctccaac ggtcccgtca tgcagaagaa gaccaagggt tgggagccta acactgagat
     5161 gatgtaccct gctgacggtg gtctcagggg ttacactgac atcgctctca aggttgacgg
     5221 tggtggccac ctccactgca acttcgtcac tacttacagg tctaagaaga ccgtcggcaa
     5281 catcaagatg cccggtgtcc acgccgttga ccaccgtctc gagaggatcg aggagtccga
     5341 caacgagacc tacgttgtcc agagggaggt cgctgttgcc aagtactcca accttggtgg
     5401 tggcatggac gagctctaca agggttctgg cggtggtggc tccggcatgc ctcagtcctg
     5461 ggaggagctc gccgctgaca agcgtgcccg tctcgctaag accatccctg acgagtggaa
     5521 ggtccagacc ctccctgccg aggactccgt tattgacttc cctaagaagt ctggcatcct
     5581 ttctgaggcc gagctcaaga tcactgaggc ctccgctgct gaccttgtct ccaagctcgc
     5641 cgccggtgag ttgacctctg tcgaggttac ccttgctttc tgtaagcgag ctgctatcgc
     5701 ccagcagttg actaactgcg cccacgagtt cttccctgac gccgctctcg cccaggctag
     5761 ggagctcgac gagtactacg ctaagcacaa gaggcccgtt ggtcctctcc acggcctccc
     5821 catctctctc aaggaccagc ttcgagtcaa gggctacgag acttctatgg gctacatctc
     5881 ttggcttaac aagtacgacg agggcgactc tgttctcact accatgctcc gtaaggccgg
     5941 tgccgtcttc tacgtcaaga cctctgtccc ccagaccctc atggtctgcg agactgtcaa
     6001 caacatcatc ggccgtaccg tcaaccctcg taacaagaac tggtcttgcg gcggctcctc
     6061 tggtggtgag ggtgccatcg ttggcattcg tggtggcgtc atcggtgttg gtaccgacat
     6121 tggtggctct attcgagtcc ccgccgcctt caacttcctc tacggtctta ggccctccca
     6181 cggccgactc ccctacgcta agatggccaa ctccatggag ggtcaggaga ccgtccactc
     6241 cgttgtcggc cccattaccc actctgttga ggacctccgt ctcttcacca agtccgtcct
     6301 cggtcaggag ccttggaagt acgactccaa ggtcatcccc atgccctggc gtcagtccga
     6361 gtctgacatt attgcctcca agatcaagaa cggcggcctc aacatcggct actacaactt
     6421 cgacggcaac gtccttcctc accctcctat cctccgtggc gtcgagacca ccgtcgccgc
     6481 tctcgccaag gccggtcaca ccgtcacccc ctggacccct tacaagcacg acttcggcca
     6541 cgacctcatc tcccacatct acgccgctga cggctccgcc gacgttatgc gtgacatctc
     6601 cgcttccggc gagcccgcca ttcctaacat caaggacctt ctcaacccca acatcaaggc
     6661 tgttaacatg aacgagctct gggacaccca cctccagaag tggaactacc agatggagta
     6721 ccttgagaag tggcgagagg ctgaggagaa ggccggcaag gaggtgagta gatcacaatt
     6781 tgtaacaggt tggatgataa gtccccggtc taacatgtag cgacgacact gaagctcact
     6841 tcttgtagct cgacgccatc atcgccccca ttacccctac cgctgccgtt cgacacgacc
     6901 agttccgata ctacggctac gcctctgtca tcaacctcct cgacttcacc tccgtcgttg
     6961 ttcccgttac cttcgccgac aagaacatcg acaagaagaa cgagtccttc aaggccgttt
     7021 ccgagcttga cgccctcgtc caggaggagt acgaccccga ggcctaccac ggcgctcccg
     7081 ttgctgtcca ggttatcggt cgaaggctct ccgaggagag gaccttggcc attgctgagg
     7141 aggtcggcaa gttgctcggt aacgtcgtca ctccttagaa ggttgagcgc gtaatagtgg
     7201 gttggtcttg tctctatcca ttcatctgtg agacatactg atttcaactt tttcatcaga
     7261 taataaggat ggtgtgagat aagcaaggct gggttatgag gtttgggact agatggtgtt
     7321 tcgcgtctaa aggtgagtct tcgttggttc aatcacattg atctgatcgc tgatcaattg
     7381 tcagcctctg aatgtgctct tgaagaaaat ggttgtctgt aagaatccag aaccaaaaac
     7441 ggtcatttca agtatgtatc ctccgtgaag ttcctttagc tcttgctaat attggacttt
     7501 agcgtgcttt agccaaagac agggtcgcga ggttaagagg atgggccagg aaaaatctaa
     7561 gttagatttg tcacacaaag aaaaagattg tctcagtttt ggttttgttt ggtttgattt
     7621 atgcatagga aatatttatg atcgaaatca ctcatttcct gcttcgtaaa tatcatctac
     7681 tatatgcttc gtaaatgtcc atctcctaca tattgtggat atgataaatg caatcctctc
     7741 gtctacactc tcggcttatg cctttgtctt ctttgcgctc cttttagcct tctttttatt
     7801 cctagtcttc aagcccatcg gtggagctcc agcttttgtt ccctttagtg agggttaatt
     7861 gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     7921 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     7981 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     8041 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     8101 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     8161 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     8221 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     8281 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     8341 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     8401 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     8461 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     8521 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     8581 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     8641 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     8701 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     8761 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     8821 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     8881 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     8941 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     9001 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     9061 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     9121 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     9181 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     9241 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     9301 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     9361 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     9421 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     9481 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     9541 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     9601 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     9661 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     9721 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     9781 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     9841 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     9901 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     9961 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
    10021 aaagtgccac
//
