LOCUS       sequence-491558-        7316 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  Plasmid for genomic integration of Atp5 (MTS)-DHFR-TEV into the 
            YORW-Delta-22 locus of yeast cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7316)
  TITLE     The IQ-compete assay measures mitochondrial protein import 
            efficiencies in living yeast cells by fluorescence dequenching
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7316)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 4, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: Atp5-DHFR-TEV (integrating)
FEATURES             Location/Qualifiers
     source          1..7316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             992..1549
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="VRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVEG
                     KQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQPE
                     LASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEYP
                     GVLSEVQEEKGIKYKFEVYEKKD"
     CDS             1556..2263
                     /codon_start=1
                     /product="tobacco etch virus protease (Kapust et al., 
                     2001)"
                     /label=TEV protease
                     /translation="GESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNK
                     HLFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQ
                     REERICLVTTNFQTKSMSSMVSDTSCTFPSSDGTFWKHWIQTKDGQCGNPLVSTRDGFI
                     VGIHSASNFTNTNNYFASVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPE
                     EPFQPVKEATQLMN"
     gene            2719..4074
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     CDS             3063..3872
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3130..3149)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3877..4074
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(join(4838..5629,5630..5698))
                     /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     5461..5480
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5699..5803)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5906..6494
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6395..6414
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgaattc gcatctagaa cggggtcatc acggctcatc atgcgccaaa caaatgtgtg
       61 caatacacgc tcggatgact gcatgatgac cgcactgact ggggacagca gatccaccta
      121 agcctgtgag agaagcagac acccgacaga tcaaggcagt tacgtctcac caaaccagat
      181 gtcaacacag ctacaacgtt gataggtcaa gatcaatgta aacaattact ttgttatgta
      241 gagttttttt agctacctat attccaccat aacatcaatc atgcggttgc tggtgtattt
      301 accaataatg tttaatgtat atatatatat atatatatgg ggccgtatac ttacatatag
      361 tagatgtcaa gcgtaggcgc ttcccctgcc ggctgtgagg gcgccataac caaggtatct
      421 atagaccgcc aatcagcaaa ctacctccgt acattcatgt tgcacccaca catttataca
      481 cccagaccgc gacaaattac ccataaggtt gtttgtgacg gcgtcgtaca agagaacgtg
      541 ggaacttttt aggctcacca aaaaagaaag aaaaaatacg agttgctgac agaagcctca
      601 agaaaaaaaa aattcttctt cgactatgct ggaggcagag atgatcgagc cggtagttaa
      661 ctatatatag ctaaattggt tccatcacct tcttttctgg tgtcgctcct tctagtgcta
      721 tttctggctt ttcctatttt tttttttcca tttttctttc tctctttcta atatataaat
      781 tctcttgcat tttctatttt tctctctatc tattctactt gtttattccc ttcaaggttt
      841 ttttttaagg agtacttgtt tttagaatat acggtcaacg aactataatt aactaaacag
      901 atctatgttt aatagagtct ttaccaggtc atttgcatca agcttaagag ctgctgcttc
      961 caaagctgct gctcccccac cggtgggatc cgttcgacca ttgaactgca tcgtcgccgt
     1021 gtcccaaaat atggggattg gcaagaacgg agatctaccc tggcctccgc tcaggaacga
     1081 gttcaagtac ttccaaagaa tgaccacaac ctcttcagtg gaaggtaaac agaatctggt
     1141 gattatgggt aggaaaacct ggttctccat tcctgagaag aatcgacctt taaaggacag
     1201 aattaatata gttctcagta gagaactcaa agaaccacca cgaggagctc attttcttgc
     1261 caaaagtttg gatgatgcct taagacttat tgaacaaccg gaattggcaa gtaaagtaga
     1321 catggtttgg atagtcggag gcagttctgt ttaccaggaa gccatgaatc aaccaggcca
     1381 cctcagactc tttgtgacaa ggatcatgca ggaatttgaa agtgacacgt ttttcccaga
     1441 aattgatttg gggaaatata aacttctccc agaataccca ggcgtcctct ctgaggtcca
     1501 ggaggaaaaa ggcatcaagt ataagtttga agtctacgag aagaaagacg gttctggaga
     1561 aagcttgttt aaggggccgc gtgattacaa cccgatatcg agcaccattt gtcatttgac
     1621 gaatgaatct gatgggcaca caacatcgtt gtatggtatt ggatttggtc ccttcatcat
     1681 tacaaacaag cacttgttta gaagaaataa tggaacactg ttggtccaat cactacatgg
     1741 tgtattcaag gtcaagaaca ccacgacttt gcaacaacac ctcattgatg ggagggacat
     1801 gataattatt cgcatgccta aggatttccc accatttcct caaaagctga aatttagaga
     1861 gccacaaagg gaagagcgca tatgtcttgt gacaaccaac ttccaaacta agagcatgtc
     1921 tagcatggtg tcagacacta gttgcacatt cccttcatct gatggcacat tctggaagca
     1981 ttggattcaa accaaggatg ggcagtgtgg caatccatta gtatcaacta gagatgggtt
     2041 cattgttggt atacactcag catcgaattt caccaacaca aacaattatt tcgcaagcgt
     2101 gccgaaaaac ttcatggaat tgttgacaaa tcaggaggcg cagcagtggg ttagtggttg
     2161 gcgattaaat gctgactcag tattgtgggg gggccataaa gttttcatgg tgaaacctga
     2221 agagcctttt cagccagtta aggaagcgac tcaactcatg aatggatcct aactcgagat
     2281 aaagcaatct tgatgaggat aatgattttt ttttgaatat acataaatac taccgttttt
     2341 ctgctagatt ttgtgatgac gtaaataagt acatattact ttttaagcca agacaagatt
     2401 aagcattaac tttacccttt tctttctaag tttcaatatt agttatcact gtttaaaagt
     2461 tatggcgaga acgtcggcgg ttaaaatata ttaccctgaa cggctgagca tgagacggaa
     2521 atctgctcgt cagtggtgct cacactgacg aatcatgtac agatcatacc gatgactgcc
     2581 tggcgactca caactaagca agacagccgg aaccagcgcc ggcgaacacc actgcatata
     2641 tggcatatca caacagtcca actagtgcac tgcagtacag tttagcttgc ctcgtccccg
     2701 ccgggtcacc cggccagcga catggaggcc cagaataccc tccttgacag tcttgacgtg
     2761 cgcagctcag gggcatgatg tgactgtcgc ccgtacattt agcccataca tccccatgta
     2821 taatcatttg catccataca ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc
     2881 tcgctccaga cctgcgagca gggaaacgct cccctcacag acgcgttgaa ttgtccccac
     2941 gccgcgcccc tgtagagaaa tataaaaggt taggatttgc cactgaggtt cttctttcat
     3001 atacttcctt ttaaaatctt gctaggatac agttctcaca tcacatccga acataaacaa
     3061 aaatgggtaa ggaaaagact cacgtttcga ggccgcgatt aaattccaac atggatgctg
     3121 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     3181 gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     3241 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     3301 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     3361 ccggcaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     3421 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     3481 acagcgatcg cgtatttcgt ctggctcagg cgcaatcacg aatgaataac ggtttggttg
     3541 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     3601 tgcataagct tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     3661 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     3721 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     3781 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     3841 agtttcattt gatgctcgat gagtttttct aaagtaactg acaataaaaa gattcttgtt
     3901 ttcaagaact tgtcatttgt atagtttttt tatattgtag ttgttctatt ttaatcaaat
     3961 gttagcgtga tttatatttt ttttcgcctc gacatcatct gcccagatgc gaagttaagt
     4021 gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat actggagtgg
     4081 accaactatc atccgctaat tactgacatt accaaatgag atctgtgaat gggcaagata
     4141 aaaaacaaaa attgaaatgt ttgacgttat gtaaaactat taattccttc gctttcggcg
     4201 gtcacagaat ttgcgtgtag ctgactcttg ttcaatcaat atcatttgtt actttatttg
     4261 aaagtctgta ttactgcgcc tattgtcatc cgtaccaaag aacgtcaaaa agaaacaaga
     4321 taatttttgt gcttacacca tttatagatc actgagccca gaatatcgct ggagctcagt
     4381 gtaagtggca tgaacacaac tctgactgat cgcacatatt gccgttatca taaatactag
     4441 ttgtacttgt caatgcgacg aatggcatca tgcctattat tacgttcctc tttttccgtt
     4501 tcatgtttcc agaatgctat tgaatctaac acttcaatta taaaaaagaa taaatccgca
     4561 ataattttag gctaattgtt gtactgtcaa gcgaacctaa tggttaaaat tcagaggaac
     4621 cttcgacgta gtctgatcgc tacttctata tcttatgttc ccagtcaatc aaaagttgat
     4681 actataatag ctgccattta tacctgttag ttatggcgat cgtttatcac gccgagcggc
     4741 cgcgattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4801 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4861 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4921 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcggg
     4981 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5041 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5101 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5161 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5221 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5281 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5341 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5401 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5461 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5521 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5581 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5641 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5701 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5761 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5821 tgccacctgt catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5881 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5941 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6001 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6061 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6121 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6181 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6241 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6301 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6361 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6421 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6481 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6541 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtgcggccg
     6601 ccaatcaccg gagcttggat atgataaacg aaatattctt gaatcgtgag atcgcctgtt
     6661 ttcaaaaccg ttggaggcag aaacaatttt gtcacaagat gggcattcta ccccatcctt
     6721 gctgtattat tgtagtctcg ctttctttta tgctggacaa atgagactac tgcacatttt
     6781 tatacgttct tggttttttt taaaggtgtg gtttcggcat tatcctgccg cacgtttctt
     6841 ggataattca tcctgattct ctattttaaa cgcttcagcc tatcaggatt tggttttgat
     6901 acatactgca agagtgtatc tcgggaacag tcatttattc cgcaacaaac ttaattgcgg
     6961 aacgcgttag gcgatttcta gcatatatca aataccgttc gcgatttctt ctgggttcgt
     7021 ctattttctt ttaaatactt attaacgtac tcaaacaact acacttcgtt gtatctcaga
     7081 atgagatccc tcagtatgac aatacatcat tctaaacgtt cgtaaaacac atatgaaaca
     7141 actttataac aaagcgaaca aaatgggcaa catgagatga aactccgcgt cccttagctg
     7201 aactacccaa acgtacgaat gcctgaacaa ttagtttaga tccgagattc cgcgcttcca
     7261 tcatttagta taatccatat tttatataat atataggata agtaacagcc cgcgaa
//
