LOCUS       sequence-491564-        7232 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for genomic integration of 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 7232)
  AUTHORS   Hoffman Y, Egeler A, Rodl S, Herrmann JM
  TITLE     The IQ-compete assay for measuring mitochondrial protein import 
            efficiencies in living yeast cells.
  JOURNAL   FEBS Lett. 2025 Oct 25. doi: 10.1002/1873-3468.70206.
  PUBMED    41137490
REFERENCE   2  (bases 1 to 7232)
  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: "FEBS Lett. 
            2025 Oct 25. doi: 10.1002/1873-3468.70206."
COMMENT     Sequence Label: DHFR-TEV (integrating)
FEATURES             Location/Qualifiers
     source          1..7232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             905..1465
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     CDS             1472..2179
                     /codon_start=1
                     /product="tobacco etch virus protease (Kapust et al., 
                     2001)"
                     /label=TEV protease
                     /translation="GESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNK
                     HLFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQ
                     REERICLVTTNFQTKSMSSMVSDTSCTFPSSDGTFWKHWIQTKDGQCGNPLVSTRDGFI
                     VGIHSASNFTNTNNYFASVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPE
                     EPFQPVKEATQLMN"
     gene            2635..3990
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     CDS             2979..3788
                     /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(3046..3065)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3793..3990
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(join(4754..5545,5546..5614))
                     /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     5377..5396
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5615..5719)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5822..6410
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6311..6330
                     /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 atctatggtt cgaccattga actgcatcgt cgccgtgtcc caaaatatgg ggattggcaa
      961 gaacggagat ctaccctggc ctccgctcag gaacgagttc aagtacttcc aaagaatgac
     1021 cacaacctct tcagtggaag gtaaacagaa tctggtgatt atgggtagga aaacctggtt
     1081 ctccattcct gagaagaatc gacctttaaa ggacagaatt aatatagttc tcagtagaga
     1141 actcaaagaa ccaccacgag gagctcattt tcttgccaaa agtttggatg atgccttaag
     1201 acttattgaa caaccggaat tggcaagtaa agtagacatg gtttggatag tcggaggcag
     1261 ttctgtttac caggaagcca tgaatcaacc aggccacctc agactctttg tgacaaggat
     1321 catgcaggaa tttgaaagtg acacgttttt cccagaaatt gatttgggga aatataaact
     1381 tctcccagaa tacccaggcg tcctctctga ggtccaggag gaaaaaggca tcaagtataa
     1441 gtttgaagtc tacgagaaga aagacggttc tggagaaagc ttgtttaagg ggccgcgtga
     1501 ttacaacccg atatcgagca ccatttgtca tttgacgaat gaatctgatg ggcacacaac
     1561 atcgttgtat ggtattggat ttggtccctt catcattaca aacaagcact tgtttagaag
     1621 aaataatgga acactgttgg tccaatcact acatggtgta ttcaaggtca agaacaccac
     1681 gactttgcaa caacacctca ttgatgggag ggacatgata attattcgca tgcctaagga
     1741 tttcccacca tttcctcaaa agctgaaatt tagagagcca caaagggaag agcgcatatg
     1801 tcttgtgaca accaacttcc aaactaagag catgtctagc atggtgtcag acactagttg
     1861 cacattccct tcatctgatg gcacattctg gaagcattgg attcaaacca aggatgggca
     1921 gtgtggcaat ccattagtat caactagaga tgggttcatt gttggtatac actcagcatc
     1981 gaatttcacc aacacaaaca attatttcgc aagcgtgccg aaaaacttca tggaattgtt
     2041 gacaaatcag gaggcgcagc agtgggttag tggttggcga ttaaatgctg actcagtatt
     2101 gtgggggggc cataaagttt tcatggtgaa acctgaagag ccttttcagc cagttaagga
     2161 agcgactcaa ctcatgaatg gatcctaact cgagataaag caatcttgat gaggataatg
     2221 attttttttt gaatatacat aaatactacc gtttttctgc tagattttgt gatgacgtaa
     2281 ataagtacat attacttttt aagccaagac aagattaagc attaacttta cccttttctt
     2341 tctaagtttc aatattagtt atcactgttt aaaagttatg gcgagaacgt cggcggttaa
     2401 aatatattac cctgaacggc tgagcatgag acggaaatct gctcgtcagt ggtgctcaca
     2461 ctgacgaatc atgtacagat cataccgatg actgcctggc gactcacaac taagcaagac
     2521 agccggaacc agcgccggcg aacaccactg catatatggc atatcacaac agtccaacta
     2581 gtgcactgca gtacagttta gcttgcctcg tccccgccgg gtcacccggc cagcgacatg
     2641 gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac
     2701 tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc catacatttt
     2761 gatggccgca cggcgcgaag caaaaattac ggctcctcgc tccagacctg cgagcaggga
     2821 aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata
     2881 aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta
     2941 ggatacagtt ctcacatcac atccgaacat aaacaaaaat gggtaaggaa aagactcacg
     3001 tttcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     3061 gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc
     3121 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     3181 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     3241 ctcctgatga tgcatggtta ctcaccactg cgatccccgg caaaacagca ttccaggtat
     3301 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     3361 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctgg
     3421 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     3481 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac
     3541 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     3601 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     3661 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     3721 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     3781 ttttctaaag taactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag
     3841 tttttttata ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt
     3901 cgcctcgaca tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc
     3961 aatcgtatgt gaatgctggt cgctatactg gagtggacca actatcatcc gctaattact
     4021 gacattacca aatgagatct gtgaatgggc aagataaaaa acaaaaattg aaatgtttga
     4081 cgttatgtaa aactattaat tccttcgctt tcggcggtca cagaatttgc gtgtagctga
     4141 ctcttgttca atcaatatca tttgttactt tatttgaaag tctgtattac tgcgcctatt
     4201 gtcatccgta ccaaagaacg tcaaaaagaa acaagataat ttttgtgctt acaccattta
     4261 tagatcactg agcccagaat atcgctggag ctcagtgtaa gtggcatgaa cacaactctg
     4321 actgatcgca catattgccg ttatcataaa tactagttgt acttgtcaat gcgacgaatg
     4381 gcatcatgcc tattattacg ttcctctttt tccgtttcat gtttccagaa tgctattgaa
     4441 tctaacactt caattataaa aaagaataaa tccgcaataa ttttaggcta attgttgtac
     4501 tgtcaagcga acctaatggt taaaattcag aggaaccttc gacgtagtct gatcgctact
     4561 tctatatctt atgttcccag tcaatcaaaa gttgatacta taatagctgc catttatacc
     4621 tgttagttat ggcgatcgtt tatcacgccg agcggccgcg attatcaaaa aggatcttca
     4681 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4741 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4801 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4861 taccatctgg ccccagtgct gcaatgatac cgcgggaccc acgctcaccg gctccagatt
     4921 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4981 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5041 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5101 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5161 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5221 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5281 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5341 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5401 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5461 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5521 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5581 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5641 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5701 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgtcatg accaaaatcc
     5761 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5821 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5881 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5941 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     6001 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6061 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6121 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6181 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6241 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6301 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6361 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6421 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6481 cgttatcccc tgattctgtg gataaccgtg cggccgccaa tcaccggagc ttggatatga
     6541 taaacgaaat attcttgaat cgtgagatcg cctgttttca aaaccgttgg aggcagaaac
     6601 aattttgtca caagatgggc attctacccc atccttgctg tattattgta gtctcgcttt
     6661 cttttatgct ggacaaatga gactactgca catttttata cgttcttggt tttttttaaa
     6721 ggtgtggttt cggcattatc ctgccgcacg tttcttggat aattcatcct gattctctat
     6781 tttaaacgct tcagcctatc aggatttggt tttgatacat actgcaagag tgtatctcgg
     6841 gaacagtcat ttattccgca acaaacttaa ttgcggaacg cgttaggcga tttctagcat
     6901 atatcaaata ccgttcgcga tttcttctgg gttcgtctat tttcttttaa atacttatta
     6961 acgtactcaa acaactacac ttcgttgtat ctcagaatga gatccctcag tatgacaata
     7021 catcattcta aacgttcgta aaacacatat gaaacaactt tataacaaag cgaacaaaat
     7081 gggcaacatg agatgaaact ccgcgtccct tagctgaact acccaaacgt acgaatgcct
     7141 gaacaattag tttagatccg agattccgcg cttccatcat ttagtataat ccatatttta
     7201 tataatatat aggataagta acagcccgcg aa
//
