LOCUS       sequence-417311-        7258 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  episomal fluorescent reporter plasmid for S. stipitis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7258)
  AUTHORS   Reichard WD, Smith SE, Robertson JB
  TITLE     BLINCAR: a reusable bioluminescent and Cas9-based genetic toolset 
            for repeatedly modifying wild-type Scheffersomyces stipitis.
  JOURNAL   mSphere. 2023 Aug 24;8(4):e0022423. doi: 10.1128/msphere.00224-23. 
            Epub 2023 Jun 22.
  PUBMED    37345937
REFERENCE   2  (bases 1 to 7258)
  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: "mSphere."; 
            date: "2023-08-24"; volume: "8(4):e0022423. doi"; pages: " 
            10.1128/msphere.00224-23. Epub 2023 Jun 22"
COMMENT     Sequence Label: pWR76
FEATURES             Location/Qualifiers
     source          1..7258
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             777..1805
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     rep_origin      complement(5503..6091)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5583..5602)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6262..7053,7054..7122))
                     /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     6885..6904
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7123..7227)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccaatgatac cggtgcgatc gtgggcccta gtacttccct cgagccgtac acttattgtt
       61 aactatgaaa catatactta tggacaaggt tcactgattt ggactgaaaa gcaaaaacga
      121 gaaatgagtc aacgtctgag atcaatccct agaccacccg ggaaggtcat gtgtttcatg
      181 aagtacgata aggttggtaa ccgattgact cattggttcg tggcggagaa gtacgcagag
      241 taaaaccggg gccgattcgt ggtaaattct ggaatgatcc agaggcgcga catttatgca
      301 gacaatttgt gttttgtcgc aaacgatgtt atagcgaaat ttttcactct gtcagataaa
      361 tggattttgt caaaaggggg aagtagaagg agaatgggcc cgagatgttc tgccaaattc
      421 tcagtagcat aatgtgaaag aagcccttac attgtccagc ctctggcatc attaaaaacc
      481 gtagcggaaa ccaattgtct ctgttcttcc ctggcacacc ctggtagccc cacccagttg
      541 tagtacatct cacacgctgg caacttggga caatcagcaa cttttttttc ttttaatttt
      601 ttcagcgcga cattttgcct cttctgcgag aacagacttt ttcacctcca tctcaccccc
      661 ctttgcactt atataaattg gaccagttcc tcccattgta gaaaaaattt tgctggacct
      721 ttttctcttt tttttgtcct ttagtttcat acaatctaag tctatctaca tgtacaatgg
      781 gtaaaaagcc tgaactcacc gcgacctctg tcgagaagtt tttgatcgaa aagttcgaca
      841 gcgtctccga cttgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg
      901 taggagggcg tggatatgtc ttgcgggtaa atagctgcgc cgatggtttc tacaaagatc
      961 gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg
     1021 gggagttcag cgagagcttg acctattgca tctcccgccg tgcacagggt gtcacgttgc
     1081 aagacttgcc tgaaaccgaa ttgcccgctg ttttgcagcc ggtcgcggag gccatggatg
     1141 cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa
     1201 tcggtcaata cactacatgg cgtgatttca tttgcgcgat tgctgatccc catgtgtatc
     1261 actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagt
     1321 tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct
     1381 ccaacaatgt cttgacggac aatggccgca taacagcggt cattgactgg agcgaggcga
     1441 tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt
     1501 gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccac
     1561 ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg
     1621 gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag
     1681 ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct
     1741 gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg
     1801 aataagcggc cgctaaccac ttgcaaaatc ctttgtattc ttgttctgca aacattttgt
     1861 ccaatctctt atctttctac gatgttgaag aacatacatt tttgttagtc cagcttgtta
     1921 tccttttttt attaaataaa aaattttatt gttaaagttg tttttcttct ttgtctgtcc
     1981 tgttttaact catatatatt ctatatatta cagaaagaaa gtagatcatc taaaataaac
     2041 atttatttga tgagacaaag taggtttttt agaatacaat gtagcatcct ctaatataca
     2101 atagagttgt tttataaaag cgatacattt aatattatga caggactggt gaagtgtata
     2161 gaattgtgct aatctggcaa acgaacccaa gaagacgtta aagcaaatag tgcattttgt
     2221 attacaatca agttcgtcga ccatgcatta gatcttgcat gcctctagaa cctaggtggc
     2281 gccacagctg tggatcctcg ttagtatttc cgtgaaggca gccattgtga aaattgaaca
     2341 tcacctgagg tattttagcc accagaagcg gcggtacgga agaaagtgtg tacgatggtt
     2401 ggtggtggaa ttgcgtgcat gcctgatggg gcaatattaa ttagatagag ctttggtgat
     2461 attagtggat aatagaattc acagagaaga catcaggagc aatttccaag agccattgat
     2521 gatgtaattg ccccaacagc aagattcaga tctgacaatt gaccaccgtt ttgtagaagc
     2581 aaaaaatcgt agattatcac caagagggtt tttcaccgaa ccagcaaata gaaactatgc
     2641 cgtagaactc gcccaggctt ttttgctagc actttccagc agtagaaccg tccaattaag
     2701 tcaacaggaa ccattgaggt cgagcccaac cacctgaacc ccctcacggt cgtgtcccta
     2761 ttattgatcc agagggtgcc agtttcggta gccaatattg gttcatgggt ttctatggcc
     2821 cggagtgagt ttgcaggttg gccccggcgc cgtctgcagg atgggagtta tagcggccaa
     2881 acttcacatt tcgaaatcct gctgcagcca atctgaagaa ttaatataaa ttcgtgtcga
     2941 atcgccgtct gtgaaatttc agtacttgat tttcttttct tcttcttttt ctcttttgtt
     3001 tcttcagaat caatacacat tttttcttcc ctataaacaa ttcatcggta ccatggtttc
     3061 aaaaggtgaa gaagataata tggctattat taaagaattt atgagattta aagttcatat
     3121 ggaaggttca gttaatggtc atgaatttga aattgaaggt gaaggtgaag gtagaccata
     3181 tgaaggtact caaactgcta aattgaaagt tactaaaggt ggtccattac catttgcttg
     3241 ggatattttg tcaccacaat ttatgtatgg ttcaaaagct tatgttaaac atccagctga
     3301 tattccagat tatttaaaat tgtcatttcc agaaggtttt aaatgggaaa gagttatgaa
     3361 ttttgaagat ggtggtgttg ttactgttac tcaagattca tcattacaag atggtgaatt
     3421 tatttataaa gttaaattga gaggtactaa ttttccatca gatggtccag ttatgcaaaa
     3481 aaaaactatg ggttgggaag cttcatcaga aagaatgtat ccagaagatg gtgctttaaa
     3541 aggtgaaatt aaacaaagat tgaagttaaa agatggtggt cattatgatg ctgaagttaa
     3601 aactacttat aaagctaaaa aaccagttca attaccaggt gcttataatg ttaatattaa
     3661 attggatatt acttcacata atgaagatta tactattgtt gaacaatatg aaagagctga
     3721 aggtagacat tcaactggtg gtatggatga attatataaa taacttaagc attttagaca
     3781 agtgcctatt tattcagatt gattttttat attgaatatg gatgttgatt acgacttgtg
     3841 ttacgaagca tgaacatgtg atttgaaaaa gtgtttacga ataaaagaac tttttttgca
     3901 accgttgcta gttgaagtgt agactctgaa tgcgaagggt tgcaaatgtg cccagtagaa
     3961 aaaatattcc atatttttct gaacttaatt caatgaaata cgagtattac tacaactgcc
     4021 aatatcggaa caatttaacg agagcaagct atataaaatg aatgttcatt tacacaagct
     4081 agaaccaaaa cttgaattaa aatttaacat aactaaatgt aactagataa aagagagaca
     4141 taaccaaacg ttcagaaggc agaagaattc agttaactcc tgcaggaatc gattggccgg
     4201 cctcacgtgg acgcgttatt aatcgtttaa acctacgtaa ccgcggctta attaaagcta
     4261 gccagcgctt gagctcagta taggatatgg tgatttagca aaatatgcct ctgttttttt
     4321 ggtaagttgt agtaattcgg aatttttcgc aacttttcca gaaatcgctt tatgccaaac
     4381 atattatacg caaattattt ataatcatgt aatatgttaa gtctctggaa tttcatgtca
     4441 gcagtgaaac gaagttgcac agtctatagc acccttaacg ctgaagtatt gactaaatgt
     4501 gtctattttt ctcttcgtac attttagtca gattgtcgaa atctaaatct ctacaggctg
     4561 caaccaaaga catcttgctg attcgtatct catttctaac tcagcaaaat tttattatcc
     4621 agcagacaac aattatctcc tttcttacat gatacttaga cttctaatct cctagtggat
     4681 aagaaaaata aatccaaaga atggaaaatg taaagctgcg ctggtgatgt ccgctaaggt
     4741 cataaactca agctacattg cgtcgaatat tttgctacga ttgctcagaa ataaaatcca
     4801 ttgtatatca tgagggtcaa taataataat aatcaacgag tccgattgct gaaccaccca
     4861 ataaatatca atcatggtct tacaagcaaa cacaacatac aataattatc ggggcggtcc
     4921 aacaatgtcg gctcgacttg tactgtatgc acctagcgtt tttcatctgg catttatccg
     4981 agtaccccta aacgctaaag cgtagagaat aatgtaacct tatcttgccg gagcaactaa
     5041 caatcgcctt gaacacaaac tgggttgctc ccctgttctc gaagtcgaag agtagctcct
     5101 tctagctcca gtgaaaatgt acagatacgc agagatggtg actgtaccta aacagactcc
     5161 gtttgcacct actccgtcag aacaatagct ctacaataaa gcctcccacc acgctccact
     5221 tacatatgca tatgcacata cattgtaaga gaagtatgtt ggccatgtag acacaaacaa
     5281 gatgtcggcg acctacagat tcttttatcc agccaatgga gagagaaggt acttgtcaga
     5341 gaagaatgca tgcaccggga aataatgcgg tgtctggggc ttttggatga gttaaaatac
     5401 ttctgacctt gtagacaccg cagagagctc catcggaaag aacatgtgag caaaaggcca
     5461 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5521 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5581 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5641 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5701 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5761 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5821 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5881 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5941 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6001 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6061 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6121 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6181 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6241 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6301 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6361 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6421 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6481 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6541 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6601 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6661 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6721 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6781 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6841 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6901 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6961 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7021 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7081 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7141 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7201 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
