LOCUS       sequence-433867-       10099 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing the RME1(ins-308A) and MKT1(30G) genes from S. 
            cerevisiae to correct sporulation defect of S288C derived yeast lab 
            strains.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10099)
  AUTHORS   Loll-Krippleber R, Jiang YK, Brown GW
  TITLE     pSPObooster: A Plasmid System to Improve Sporulation Efficiency of 
            Saccharomyces cerevisiae Lab Strains.
  JOURNAL   Yeast. 2024 Sep 9. doi: 10.1002/yea.3978.
  PUBMED    39248173
REFERENCE   2  (bases 1 to 10099)
  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: "Yeast. 2024
            Sep 9. doi: 10.1002/yea.3978."
COMMENT     Sequence Label: pSPObooster
FEATURES             Location/Qualifiers
     source          1..10099
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     73..95
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     87..104
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     88..104
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     111..130
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        111..129
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5945..5963)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5984..6000)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5984..6000)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5997..6019)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6008..6024
                     /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)."
     primer_bind     complement(6334..6351)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6505..7093)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6585..6604)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7272..8063,8064..8132))
                     /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     7895..7914
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8133..8237)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    8280..8783
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(9101..9904)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDLLTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSACVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLATGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(join(9905..10099,1..22))
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
ORIGIN
        1 aaaaaaaaaa agatgaatga aaagcgatcg gatgtgctgc aaggcgatta agttgggtaa
       61 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg taatacgact
      121 cactataggg cgaattggag aaacagaata atatataaac ctgtataata taaccttgaa
      181 gactatattt cttttcgaag taggaagaat gactgagcga aaaaaaccgt agaagtagaa
      241 cttaaaataa gaacggaggc taaaccacca aaacagctca tcaacagtta aaacaccacc
      301 aatatttcca gaaaaaagaa ctgtagatat agtctcatta gtatttagaa gaaagaggat
      361 tttgcaatgg caggggatag tataagcgct gacggcactg gcgtccatcc agttgaactg
      421 tcggtgtatt cagtattgtc caccgattta gacggactgt atcaatcgat caatgaatta
      481 cgcgagtcgc aagcactact catccttatg cttcggaagg ttcgcgataa acttcgaaga
      541 gaaggtcaag tcctttatga cccagaacca ttcaaaccaa ccatggataa actggctgac
      601 ttatctgccc gtgtgcgcat tctctcacaa agatacgagg agcttcaggg gaatgctaga
      661 gccctcaaca actaagtaaa catacacatg tatagcataa tcttcgattc cttttaccca
      721 ttcgtagatg gcatcgacag tgtacttata atactaccgt gagaaacgat attaagaaca
      781 agacgaaaaa tggtggaaaa tctggaaagc aagaacgtag cacgctagag ttattctagc
      841 ttgtcagctc gactgtttaa ttattattgt tattatatta acattaacgt aataaccatt
      901 gaactacatt aaagaaataa ctgtctcgct tatatctggt ctattttttg ttttttgcct
      961 ttccttttaa attctttata accttgaaaa caaaaaatca cattggtaaa ttacaatcat
     1021 ttattaccaa atttgtccat taactatttt agcgtgactg actgataaag actgtttcac
     1081 tctctttatt tattcataac aaaaattcag atattttgaa ctataaagta ctaaaggcag
     1141 aaaaattaat agcaaattaa gcgatggcaa tcaagtcatt ggaatcgttc cttttcgaaa
     1201 gaggtctagt aggatcctat gccattgagg ctctgaataa ttgtaccctg ggtatagacg
     1261 tcaaccatta tgtttccaga ttgttgacca ataaaagaga acagtatttg gatgccattg
     1321 gtggtttccc taccagttta aaaatgtatt tggaaagcga cttaaagata ttcaaggatt
     1381 tcaatattac ccctattttt gtctttaatg gtggtttgac ttacaaccaa ttagaggcat
     1441 cgggccattt tactgcagca tctgcttctg cgtcgatttc gtcaaccact acttccagca
     1501 gcggtaccaa tgccacaaca agatccaaca cggaatccgt tcttttgcag agaagtagag
     1561 gctggacaca atggaacaac ttgattagtt cgaatcagaa ttcctatatt gaccagccta
     1621 ttcaacccca agagccattc agacataaca ccacaattga ctccaaagct taccaaaacg
     1681 atttgatcgc ttattttatt gagcatgggt atatgtacca agtggctcct tactcctctt
     1741 ggtttcaact agcatattta ttgaatagcg cttatattga tgcgatctac ggcccaactg
     1801 attgtttgat gctagattgt gttgataggt tcattttggg tatggaattt ccaaataaag
     1861 aatttcgatt cattgacagg tcaagagtga tgaaggattt aggttgtacc catgaagagt
     1921 ttatcgatat tgctatggct gtaggtaatg atttgcaacc aacaacttta ccaccattgc
     1981 aaatttatcc agttcctcag ctgtttgata ttgctctaga aatggttttg aatacaggta
     2041 caaacttcta tgcctatcaa ctatctacaa ccttacaaaa tgactccaaa gagaatattc
     2101 aaaactacca aagaggtatt tctgcattaa gatacatgcc agtcttgaaa gatacaggta
     2161 aagttgaatt atttgttcaa gagatcgttg tttcagagga ggacagtgaa aagaataaca
     2221 aggatggaaa aaagagtaat ctttcctcac catcttctgc gtcatctagt gcttctcctg
     2281 ctaccaccgt gactaagaac gctagtgaaa agttaactta tgaaaaatcc agcactaaag
     2341 aggttagaaa accaagagat attcctaatg atgttcatga ttttatcggc caaatgttac
     2401 ctcatgaata ctatttctat agatccatag gactggttac tggaaaatta tttgacgcaa
     2461 ttgtcacagg agtttatcct gaagagcctc cattgggcgg aggatcttct acatcctaca
     2521 agaaactggt ttccaagtcc gttgaaatct tcaagaataa agaaatcaat ttattaactc
     2581 aaccaattaa cagatactat cagattaaac aaattaaaca agtcaagtgg tatgcggcaa
     2641 atgagcctac cactttgacc aacaggatgt caccttctat gtttgaaact atcaaccact
     2701 taattgttaa aacagaaacg tctgatgaaa aggaattttc tatctctgag ttcattacaa
     2761 caataaatgg ttctagcaat atggcgaaag attttatatc tgaaaaagtc attttcccaa
     2821 actccgtacc aatcgaatcc aaattaaact caccatttaa tttattgtcg accaatttct
     2881 taagattact agtactattg gagttcttca cttttgattt caaggaaaaa ctgctagagc
     2941 ctacgagatg gggtgaggta ttcttaaaat taaatgagct caatattgac agtaagtatc
     3001 acgaatcagt tattatcttt ttggtatttt tgaaatgcga tgtcttaaaa cttgacgaag
     3061 aagttcaacc accggctccc tcagccctat ctcaagccac tcttcgttca tatcctgaag
     3121 aatcattgta cgtcctattg attacgcgtg tattaacttt atttcaagtg gaccaaaagc
     3181 catcaaatta ccacggacca attgataaga aaactttgat ctttagagat catctttctt
     3241 tcataaagga aaatttaaat gaactttttg aagccgtttt gatctcatct ttaacttctg
     3301 gcgagtttaa tagattatca cttgataact ttggttgggc aagaaagatt gttagatatt
     3361 taccgttcaa attagattcc ccaaatacta taatggctat gatgtgggag tttttcttac
     3421 aaaaatatct tcacaatggt aatgccaaga atgatgcatt gtctttagtt gctactgaat
     3481 tcaacactta caaatccaca ccaaatcttg atgaacagtt tgtcgaatcc catagatttc
     3541 ttcttgaaat ttcaaaagtc atgcaggagc taaatgctgc gaaattaatc gatgaaaacg
     3601 tgtttaaatt attcactaaa gcagttgagt ttactacaac agctctatct agctgaagcg
     3661 tgagaatgag tatgatggaa atagtggaac tatttaaaaa gcacccaata aagaaagata
     3721 gttactgtaa agaggtatat tatcattatt cagagggttc aagtaccatc atcaagattt
     3781 aacacaagca cagtgaatag caacaattaa agaaaaaact aatcaaaatc acggtagaaa
     3841 taggagctgt ccaaagaagc aaagtgaata tagaagaaaa aatgcgagta cttacgattt
     3901 tttatttagc taagtaacca acgtggtgcc atattcacga catagcaagt gtccatagtt
     3961 gcatgcgttt tctttaatat cattgtgttt tatttttatt actttaagaa aaaatacagt
     4021 agctttcatg cctgcacaaa tttaacaaga gtttcatggg gtacattttt aatgcctcaa
     4081 ctatttggta ttgttcaaaa ttttcttcaa acgcttgtta aatcttgaat cagcgttgtt
     4141 atgcaccatg gccacatgtc tcttataggc gtctttgcgc ctaaacgttt tctgacagat
     4201 aggaaatggg caattcagtc ctggatattt ttctactttt agatttaatg atttctccat
     4261 ctctatatca agctctccta tatgttggga agcacaatgt cttctcagac cagttgcttg
     4321 ttggaaaccc aatattttcc agggacattg ctctatggga cacttacacg gtctttcaag
     4381 gttgaattcg tctaagtgcg cggcaaactc aactaacgtt gcgaactttt cagaacaatg
     4441 agaacagtgg taacttcctc tttttgtttg cccttcggaa attgtgctga gtactttggg
     4501 gtcgtgatat agctgtgcca cgtgcgattt ttggggatag cctagaatac actgtgaatg
     4561 ggattgttct gttgtattgc tctcaataaa tacaggattt gctccttttt gacgtccata
     4621 attaataaga tctctgaact ccggatcaac taatgacgtg atacagtgtg tactgtccgc
     4681 tatttcatcc ggccgtaatt gaggtgctgt accttcctca gtagacgacg tagagtcata
     4741 agcctgtctc tgatttccgg ctataactgt gattgtgttt tctaatgata gtagtaaaca
     4801 gtcctcagtg ctggaaggca ggctggaagg caggctggaa tccccctcta agggtgatgc
     4861 cgaatattct ttcatgtttt gcccgaaatc gtgccaatga taaatcggtt gcacctcttg
     4921 taaaacctct ctgttccaag accccttggc gatggcactg ttttgtccat aacacggtga
     4981 cataagtgct gaaaatccca acttctagta aagaattaac aaattcaact gtatgactta
     5041 taggataaga acaatgtcag ttccaatgcg ttgacacagt agaaaatata tttactaaga
     5101 agcagcttcg tcgttggtat agtttcggtt gatctagata taaatataac tacaaataaa
     5161 tggcaggaaa aattgaagca aatgacacag gacatatcta tcacattgct ccctttaatt
     5221 tcctcgaagg cactctggcc tttgttctct gcataagaaa aagccagcag aaaatacggt
     5281 gatatattta attacatgcc cggccacttt tctgctggta aagcccaccg gaaaagttcg
     5341 gattttctta tccagtatac gccgaccttt tgatgtaatc tgtgacatct cactccggcg
     5401 gccattttac gtgacgaagc atcccttaca acagaaagaa gaaaaaagat atgccgcttt
     5461 gcggtttctt tctggcaatg tatgcactca taatgctact cgtttaccca ctatccctgt
     5521 ccaaactaaa gagggaggaa agcacttttt gcatttacac atcgtagatt ataaaatgat
     5581 cgttaacagg cgcttgtgat tttgaattta agaaatgtgg actagagaag tcttaaatcg
     5641 ccaatgctgt accagactct ctatagcatc taaacacgaa attcaactgt tatcttagtt
     5701 tttcacttac cagtagcgcg cttgttattc ccacgttatt atttgccccc acatcatagg
     5761 tcaagtgacc ttctcttacc cgacatgaat aaagaaaaga aaagaaatca tacccttcag
     5821 cctgtttagc cataaatagt aaagagtaga tgtttcgacg gactaaataa tgtgatattt
     5881 acagataacg atgtcgttat tatatatata tatatatatt gtcacttgta cccagctttt
     5941 gttcccttta gtgagggtta atttcgagct tggcgtaatc atggtcatag ctgtttcctg
     6001 tgtgaaattg ttatccgctc acaacagctg gatgaattga atagctttat ggaccctgaa
     6061 accacagcca cattaacctt ctttgatggt caaaacttat ccttcaccat aaatatgcct
     6121 cgcaaaaaag gtaattaaca tatatagaat tacattattt atgaaatatc atcactatct
     6181 cttagcatct ttaatccttt tctacatcag ataacttcgg tttgttatca tcgtctgtat
     6241 tgtcatcaat tggcgcgcct ctccctgcag gcattaatga atcggccaac gcgcggggag
     6301 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6361 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6421 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6481 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6541 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6601 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6661 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6721 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6781 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6841 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6901 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6961 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7021 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7081 aaaaggatct caagaagatc ctttgatctt ttgtttaaac ctacggggtc tgacgctcag
     7141 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7201 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7261 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7321 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7381 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7441 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7501 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7561 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7621 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7681 tgcaaaaaag cggttagctc cttcggtcct ccgatggttg tcagaagtaa gttggccgca
     7741 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7801 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7861 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7921 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7981 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8041 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8101 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     8161 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8221 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcgg gtccttttca
     8281 tcacgtgcta taaaaataat tataatttaa attttttaat ataaatatat aaattaaaaa
     8341 tagaaagtaa aaaaagaaat taaagaaaaa atagtttttg ttttccgaag atgtaaaaga
     8401 ctctaggggg atcgccaaca aatactacct tttatcttgc tcttcctgct ctcaggtatt
     8461 aatgccgaat tgtttcatct tgtctgtgta gaagaccaca cacgaaaatc ctgtgatttt
     8521 acattttact tatcgttaat cgaatgtata tctatttaat ctgcttttct tgtctaataa
     8581 atatatatgt aaagtacgct ttttgttgaa attttttaaa cctttgttta tttttttttc
     8641 ttcattccgt aactcttcta ccttctttat ttactttcta aaatccaaat acaaaacata
     8701 aaaataaata aacacagagt aaattcccaa attattccat cattaaaaga tacgaggcgc
     8761 gtgtaagtta caggcaagcg atccgtctta attaactctg gcgcgccact actttttgga
     8821 actcttgttg ttctttggag ttcaatgcgt ccatctttac agtcctgtct tattgttctt
     8881 gatttgtgcc ccgtaaaata ctgttacttg gttctggcga ggtattggat agttcctttt
     8941 tataaaggcc atgaaggttt ttctttccaa tttttttttt ttcgtcatta tagaaatcat
     9001 tacgaccgag attcccggca tatggtaata actgatataa ttaaattgaa gctctaattt
     9061 gtgagtttag tatacatgca tttacttata atacagtttt ttagttttgc tggccgcatc
     9121 ttctcaaata tgcttcccag cctgcttttc tgtaacgttc accctctacc ttagcatccc
     9181 ttccctttgc aaatagtcct cttccaacaa taataatgtc agatcctgta gagaccacat
     9241 catccacggt tctatactgt tgacccaatg cgtctccctt gtcatctaaa cccacaccgg
     9301 gtgtcataat caaccaatcg taaccttcat ctcttccacc catgtctctt tgagcaataa
     9361 agccgataac aaaatctttg tcgctcttcg caatgtcaac agtaccctta gtatattctc
     9421 cagtagctag ggagcccttg catgacaatt ctgctaacat caaaaggcct ctaggttcct
     9481 ttgttacttc ttccgccgcc tgcttcaaac cgctaacaat acctgggccc accacaccgt
     9541 gtgcattcgt aatgtctgcc cattctgcta ttctgtatac acacgcagag tactgcaatt
     9601 tgactgtatt accaatgtca gcaaattttc tgtcttcgaa gagtaaaaaa ttgtacttgg
     9661 cggataatgc ctttagcggc ttaactgtgc cctccatgga aaaatcagtc aagagatcca
     9721 catgtgtttt tagtaaacaa attttgggac ctaatgcttc aactaactcc agtaattcct
     9781 tggtggtacg aacatccaat gaagcacaca agtttgtttg cttttcgtgc atgatattaa
     9841 atagcttggc agcaacagga ctaggatgag tagcagcacg ttccttatat gtagctttcg
     9901 acatgattta tcttcgtttc ctgcacgttt ttgttctgtg cagttgggtt aagaatactg
     9961 ggcaatttca tgtttcttca acaccaccta tgcgtatata taccaatcta agtctgtgct
    10021 ccttccttcg ttcttccttc tgctcggaga ttaccgaatc aaaaaaattt caaagaaacc
    10081 ggaatcaaaa aaaagaaca
//
