LOCUS       Exported                6430 bp ds-DNA     circular SYN 18-SEP-2025
DEFINITION  Plasmid for tagging Kre2 with moxGFP2 by gene replacement.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6430)
  AUTHORS   Glick Lab
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Oct 2, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Linearize with BlpI for integration at KRE2.
FEATURES             Location/Qualifiers
     source          1..6430
                     /organism="synthetic DNA construct"
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             245..1211
                     /codon_start=2
                     /label=KRE2
                     /translation="PKACYVTLVRNKELKGLLSSIKYVENKINKKFPYPWVFLNDEPFT
                     EEFKEAVTKAVSSEVKFGILPKEHWSYPEWINQTKAAEIRADAATKYIYGGSESYRHMC
                     RYQSGFFWRHELLEEYDWYWRVEPDIKLYCDINYDVFKWMQENEKVYGFTVSIHEYEVT
                     IPTLWQTSMDFIKKNPEYLDENNLMSFLSNDNGKTYNLCHFWSNFEIANLNLWRSPAYR
                     EYFDTLDHQGGFFYERWGDAPVHSIAAALFLPKDKIHYFSDIGYHHPPYDNCPLDKEVY
                     NSNNCECDQGNDFTFQGYSCGKEYYDAQGLVKPKNWKKFRE"
     CDS             1221..1913
                     /codon_start=1
                     /product="oxidation-resistant GFP derivative (Valbuena et 
                     al., 2024)"
                     /label=moxGFP2
                     /note="yeast codon-optimized"
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FISTTGKLPVPWPTLVTTLTYGVQVFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGIT"
     3'UTR           1914..2904
                     /label=3' KRE2
     primer_bind     complement(2923..2939)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3407..4210)
                     /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
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(4211..4431)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     promoter        4524..4628
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4629..5489
                     /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"
     rep_origin      5660..6248
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 atgcgccaaa agcttgttac gtcactttgg tgagaaacaa ggagttgaaa ggtttgctaa
      301 gctccattaa atatgtggaa aacaaaatta acaagaaatt cccatatcct tgggttttcc
      361 taaacgatga accttttact gaagaattca aggaagcagt caccaaagct gtttcttccg
      421 aagttaagtt tggtattttg cccaaggaac attggtcata tcctgaatgg attaatcaaa
      481 ccaaggctgc tgaaattcgt gcagatgctg ccaccaaata catatacggt ggctccgaat
      541 cttatagaca catgtgtcgt taccaatctg ggtttttctg gagacatgaa ttattagaag
      601 agtacgattg gtactggcgt gtggaaccag acatcaagtt atactgtgat attaattacg
      661 acgtttttaa gtggatgcaa gaaaacgaaa aagtttacgg ctttaccgtt tctattcatg
      721 aatatgaagt gacgatccca acactatggc aaacgtccat ggatttcatc aaaaagaacc
      781 ccgaatactt agatgaaaac aacctgatga gttttctttc gaacgataat ggtaaaacat
      841 acaatctgtg ccatttctgg tcaaactttg aaattgcaaa cttgaatttg tggaggtcac
      901 cagcctacag agagtatttt gacactttgg atcatcaagg tggatttttc tacgaaagat
      961 ggggcgatgc tcccgttcat tctattgctg ctgctttgtt tttgccaaag gataaaatcc
     1021 attatttttc agacattggt taccatcatc caccttatga taactgccca ttggacaagg
     1081 aggtctataa cagtaacaac tgtgaatgtg accaaggtaa tgatttcact ttccaaggtt
     1141 actcttgtgg taaggaatat tatgatgctc aagggttggt aaagccaaaa aactggaaaa
     1201 aattccgtga gggaagttcg atggatagca ctgagagctt gttcacaggt gttgttccta
     1261 tcttggtcga attagacggt gacgtaaacg gtcataagtt cagtgttaga ggtgaaggtg
     1321 aaggtgacgc tactaacggt aaattgacat tgaagtttat tagtactaca ggtaaattgc
     1381 cagttccttg gccaactttg gttactacat tgacatacgg tgttcaagtt ttttcaagat
     1441 acccagatca tatgaagcaa catgatttct ttaagtctgc tatgccagaa ggttacgttc
     1501 aagaaagaac tatcactttt aaagatgatg gtacttacaa gacaagagca gaggttaagt
     1561 tcgaaggtga cactttggtt aacagaatcg aattgaaggg tatcgatttt aaagaagatg
     1621 gtaatatttt gggtcataaa ttggaataca actacaactc acataacgtt tacattacag
     1681 ctgataaaca aaagaatggt attaaagcaa acttcaagat cagacataat gttgaagatg
     1741 gttctgttca attagcagat cattaccaac aaaacactcc aattggtgac ggtccagttt
     1801 tgttaccaga taaccattac ttgtctacac aatcaaaatt gtctaaagat ccaaacgaaa
     1861 agagagatca tatggttttg ttagaatttg ttactgctgc tggtatcaca taaaaatctt
     1921 ggaacatact gtttctttgt tttgacttta tactttctat ttatatttta tttttataac
     1981 tggttaagta cacataggac tgcgtatcaa acatataagt gaggcaatcc acattttttt
     2041 taaagattcg aatattttta ttctcattag cgtattccga gaatagttcg aaaaaatata
     2101 aggtatatca agagttttta caagtgagag gaaagaggaa taagctataa gcaacaaaag
     2161 cgtaaaaaaa ttagctgaag acatagaact atggatgttc tcaaagaggt gttgtcacta
     2221 gaccaagata aatttgacca gctgaaggaa acgagccgag ataaaacaaa tgaaacggat
     2281 gatccttttg aaaactattt gaaggattgt aaatttaaag cgccttcaaa caaagatcag
     2341 tcaccatttg ctaaacttaa atcattacag gaaactcatt ctaacaatga agcggctatt
     2401 aatataatta ttcctcaatt gattgattac ttaaccgaat tcactaatag gttatcaaat
     2461 tacacacaag atttagactt cattaaaaaa aagtccaatg aattacagtc attgctcgaa
     2521 tacaactcca ctaaactggc acatatctct cctatggtta atgatttgat gattcctcct
     2581 gaactcattg atgacatcat taaagggaag atcaatgaaa gctggcagga taatataaca
     2641 ttcatagcag ataaagaaga aatttataac aagtataggt ccaataatct cgatcaagac
     2701 aacaaggacg cagaaaattc agcaatgcta gcaccaaagg attttgataa gttatgtcaa
     2761 ctcctggaca tcctaaaaaa tgttattcta gaaaggtcga aaagacttat tatttccaaa
     2821 atcaaaactt tgaggagtca taacccagta ccatcgcaaa ggatacaaaa caaattatta
     2881 aaagttcaaa aaattttccc cttcggtacc gagctcgaat tcactggccg tcgttttaca
     2941 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     3001 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     3061 cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3121 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3181 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     3241 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     3301 atcaccgaaa cgcgcgagac gaaagggcgg gtaataactg atataattaa attgaagctc
     3361 taatttgtga gtttagtata catgcattta cttataatac agttttttag ttttgctggc
     3421 cgcatcttct caaatatgct tcccagcctg cttttctgta acgttcaccc tctaccttag
     3481 catcccttcc ctttgcaaat agtcctcttc caacaataat aatgtcagat cctgtagaga
     3541 ccacatcatc cacggttcta tactgttgac ccaatgcgtc tcccttgtca tctaaaccca
     3601 caccgggtgt cataatcaac caatcgtaac cttcatctct tccacccatg tctctttgag
     3661 caataaagcc gataacaaaa tctttgtcgc tcttcgcaat gtcaacagta cccttagtat
     3721 attctccagt agatagggag cccttgcatg acaattctgc taacatcaaa aggcctctag
     3781 gttcctttgt tacttcttct gccgcctgct tcaaaccgct aacaatacct gggcccacca
     3841 caccgtgtgc attcgtaatg tctgcccatt ctgctattct gtatacaccc gcagagtact
     3901 gcaatttgac tgtattacca atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt
     3961 acttggcgga taatgccttt agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga
     4021 tatccacatg tgtttttagt aaacaaattt tgggacctaa tgcttcaact aactccagta
     4081 attccttggt ggtacgaaca tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga
     4141 tattaaatag cttggcagca acaggactag gatgagtagc agcacgttcc ttatatgtag
     4201 ctttcgacat gatttatctt cgtttcctgc atgtttttgt tctgtgcagt tgggttaaga
     4261 atactgggca atttcatgtt tcttcaacac tacatatgcg tatatatacc aatctaagtc
     4321 tgtgctcctt ccttcgttct tccttctgtt cggagattac cgaatcaaaa aaatttcaaa
     4381 gaaaccgaaa tcaaaaaaaa gaataaaaaa aaaatgatga attgaattga aaagctggcc
     4441 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     4501 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4561 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4621 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4681 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4741 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4801 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4861 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4921 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4981 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     5041 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5101 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5161 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     5221 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5281 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5341 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5401 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5461 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     5521 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5581 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5641 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5701 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5761 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5821 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5881 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5941 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6001 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6061 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6121 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6181 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6241 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6301 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     6361 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     6421 aagcggaaga
//
