LOCUS       sequence-491557-        7391 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  Plasmid for genomic integration of Oxa1(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 7391)
  TITLE     The IQ-compete assay measures mitochondrial protein import 
            efficiencies in living yeast cells by fluorescence dequenching
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7391)
  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            "Unpublished"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: Oxa1-DHFR-TEV (integrating)
FEATURES             Location/Qualifiers
     source          1..7391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1067..1624
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="VRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVEG
                     KQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQPE
                     LASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEYP
                     GVLSEVQEEKGIKYKFEVYEKKD"
     CDS             1631..2338
                     /codon_start=1
                     /product="tobacco etch virus protease (Kapust et al., 
                     2001)"
                     /label=TEV protease
                     /translation="GESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNK
                     HLFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQ
                     REERICLVTTNFQTKSMSSMVSDTSCTFPSSDGTFWKHWIQTKDGQCGNPLVSTRDGFI
                     VGIHSASNFTNTNNYFASVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPE
                     EPFQPVKEATQLMN"
     gene            2794..4149
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     CDS             3138..3947
                     /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(3205..3224)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3952..4149
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(join(4913..5704,5705..5773))
                     /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     5536..5555
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5774..5878)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5981..6569
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6470..6489
                     /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 atctatgttc aaactcacct ctcgactcgt cacgtcaagg tttgctgcct cttccagact
      961 ggccaccgct cgaaccatag tattgccccg gccccatccg tcatggatct cttttcaggc
     1021 caaaagattt aattcgacgg gcccaaatgc caacgatgtc ggatccgttc gaccattgaa
     1081 ctgcatcgtc gccgtgtccc aaaatatggg gattggcaag aacggagatc taccctggcc
     1141 tccgctcagg aacgagttca agtacttcca aagaatgacc acaacctctt cagtggaagg
     1201 taaacagaat ctggtgatta tgggtaggaa aacctggttc tccattcctg agaagaatcg
     1261 acctttaaag gacagaatta atatagttct cagtagagaa ctcaaagaac caccacgagg
     1321 agctcatttt cttgccaaaa gtttggatga tgccttaaga cttattgaac aaccggaatt
     1381 ggcaagtaaa gtagacatgg tttggatagt cggaggcagt tctgtttacc aggaagccat
     1441 gaatcaacca ggccacctca gactctttgt gacaaggatc atgcaggaat ttgaaagtga
     1501 cacgtttttc ccagaaattg atttggggaa atataaactt ctcccagaat acccaggcgt
     1561 cctctctgag gtccaggagg aaaaaggcat caagtataag tttgaagtct acgagaagaa
     1621 agacggttct ggagaaagct tgtttaaggg gccgcgtgat tacaacccga tatcgagcac
     1681 catttgtcat ttgacgaatg aatctgatgg gcacacaaca tcgttgtatg gtattggatt
     1741 tggtcccttc atcattacaa acaagcactt gtttagaaga aataatggaa cactgttggt
     1801 ccaatcacta catggtgtat tcaaggtcaa gaacaccacg actttgcaac aacacctcat
     1861 tgatgggagg gacatgataa ttattcgcat gcctaaggat ttcccaccat ttcctcaaaa
     1921 gctgaaattt agagagccac aaagggaaga gcgcatatgt cttgtgacaa ccaacttcca
     1981 aactaagagc atgtctagca tggtgtcaga cactagttgc acattccctt catctgatgg
     2041 cacattctgg aagcattgga ttcaaaccaa ggatgggcag tgtggcaatc cattagtatc
     2101 aactagagat gggttcattg ttggtataca ctcagcatcg aatttcacca acacaaacaa
     2161 ttatttcgca agcgtgccga aaaacttcat ggaattgttg acaaatcagg aggcgcagca
     2221 gtgggttagt ggttggcgat taaatgctga ctcagtattg tgggggggcc ataaagtttt
     2281 catggtgaaa cctgaagagc cttttcagcc agttaaggaa gcgactcaac tcatgaatgg
     2341 atcctaactc gagataaagc aatcttgatg aggataatga tttttttttg aatatacata
     2401 aatactaccg tttttctgct agattttgtg atgacgtaaa taagtacata ttacttttta
     2461 agccaagaca agattaagca ttaactttac ccttttcttt ctaagtttca atattagtta
     2521 tcactgttta aaagttatgg cgagaacgtc ggcggttaaa atatattacc ctgaacggct
     2581 gagcatgaga cggaaatctg ctcgtcagtg gtgctcacac tgacgaatca tgtacagatc
     2641 ataccgatga ctgcctggcg actcacaact aagcaagaca gccggaacca gcgccggcga
     2701 acaccactgc atatatggca tatcacaaca gtccaactag tgcactgcag tacagtttag
     2761 cttgcctcgt ccccgccggg tcacccggcc agcgacatgg aggcccagaa taccctcctt
     2821 gacagtcttg acgtgcgcag ctcaggggca tgatgtgact gtcgcccgta catttagccc
     2881 atacatcccc atgtataatc atttgcatcc atacattttg atggccgcac ggcgcgaagc
     2941 aaaaattacg gctcctcgct ccagacctgc gagcagggaa acgctcccct cacagacgcg
     3001 ttgaattgtc cccacgccgc gcccctgtag agaaatataa aaggttagga tttgccactg
     3061 aggttcttct ttcatatact tccttttaaa atcttgctag gatacagttc tcacatcaca
     3121 tccgaacata aacaaaaatg ggtaaggaaa agactcacgt ttcgaggccg cgattaaatt
     3181 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
     3241 gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
     3301 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg
     3361 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     3421 tcaccactgc gatccccggc aaaacagcat tccaggtatt agaagaatat cctgattcag
     3481 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt
     3541 gtaattgtcc ttttaacagc gatcgcgtat ttcgtctggc tcaggcgcaa tcacgaatga
     3601 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     3661 aagtctggaa agaaatgcat aagcttttgc cattctcacc ggattcagtc gtcactcatg
     3721 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     3781 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     3841 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     3901 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaaagt aactgacaat
     3961 aaaaagattc ttgttttcaa gaacttgtca tttgtatagt ttttttatat tgtagttgtt
     4021 ctattttaat caaatgttag cgtgatttat attttttttc gcctcgacat catctgccca
     4081 gatgcgaagt taagtgcgca gaaagtaata tcatgcgtca atcgtatgtg aatgctggtc
     4141 gctatactgg agtggaccaa ctatcatccg ctaattactg acattaccaa atgagatctg
     4201 tgaatgggca agataaaaaa caaaaattga aatgtttgac gttatgtaaa actattaatt
     4261 ccttcgcttt cggcggtcac agaatttgcg tgtagctgac tcttgttcaa tcaatatcat
     4321 ttgttacttt atttgaaagt ctgtattact gcgcctattg tcatccgtac caaagaacgt
     4381 caaaaagaaa caagataatt tttgtgctta caccatttat agatcactga gcccagaata
     4441 tcgctggagc tcagtgtaag tggcatgaac acaactctga ctgatcgcac atattgccgt
     4501 tatcataaat actagttgta cttgtcaatg cgacgaatgg catcatgcct attattacgt
     4561 tcctcttttt ccgtttcatg tttccagaat gctattgaat ctaacacttc aattataaaa
     4621 aagaataaat ccgcaataat tttaggctaa ttgttgtact gtcaagcgaa cctaatggtt
     4681 aaaattcaga ggaaccttcg acgtagtctg atcgctactt ctatatctta tgttcccagt
     4741 caatcaaaag ttgatactat aatagctgcc atttatacct gttagttatg gcgatcgttt
     4801 atcacgccga gcggccgcga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4861 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4921 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4981 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5041 caatgatacc gcgggaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5101 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5161 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5221 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5281 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5341 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5401 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5461 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5521 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5581 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5641 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5701 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5761 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5821 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5881 catttccccg aaaagtgcca cctgtcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5941 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6001 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6061 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6121 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6181 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6241 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6301 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6361 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6421 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6481 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6541 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6601 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6661 ataaccgtgc ggccgccaat caccggagct tggatatgat aaacgaaata ttcttgaatc
     6721 gtgagatcgc ctgttttcaa aaccgttgga ggcagaaaca attttgtcac aagatgggca
     6781 ttctacccca tccttgctgt attattgtag tctcgctttc ttttatgctg gacaaatgag
     6841 actactgcac atttttatac gttcttggtt ttttttaaag gtgtggtttc ggcattatcc
     6901 tgccgcacgt ttcttggata attcatcctg attctctatt ttaaacgctt cagcctatca
     6961 ggatttggtt ttgatacata ctgcaagagt gtatctcggg aacagtcatt tattccgcaa
     7021 caaacttaat tgcggaacgc gttaggcgat ttctagcata tatcaaatac cgttcgcgat
     7081 ttcttctggg ttcgtctatt ttcttttaaa tacttattaa cgtactcaaa caactacact
     7141 tcgttgtatc tcagaatgag atccctcagt atgacaatac atcattctaa acgttcgtaa
     7201 aacacatatg aaacaacttt ataacaaagc gaacaaaatg ggcaacatga gatgaaactc
     7261 cgcgtccctt agctgaacta cccaaacgta cgaatgcctg aacaattagt ttagatccga
     7321 gattccgcgc ttccatcatt tagtataatc catattttat ataatatata ggataagtaa
     7381 cagcccgcga a
//
