LOCUS       sequence-491566-        7040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for genomic integration of the IQ-compete reporter 
            (GFP-TCS-Quencher-CL1) into the YPRC-Delta-15 locus of yeast cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7040)
  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 7040)
  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: IQ-compete reporter (integrating)
FEATURES             Location/Qualifiers
     source          1..7040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        472..885
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             905..1618
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(935..963)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1558..1578
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1625..1645
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     gene            2247..3821
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     CDS             2591..3619
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             2591..3619
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      3624..3821
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(join(4583..5374,5375..5443))
                     /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     5206..5225
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5444..5548)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5651..6239
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6140..6159
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgaattc gcatctagat ggtagagcca caaacagccg gtacaagcaa cgatctccag
       61 gaccatctga atcatgcgcg gatgacacga actcacgacg gcgatcacag acattaaccc
      121 acagtacaga cactgcgaca acgtggcaat tcgtcgcaat accgtctcac tgaactggcc
      181 gataattgca gacgaacgcc ttgccaacag ggagttcttc agagacatgg aggctcaaaa
      241 cgaaattatt gacagcctag acatcaatag tcatacaaca gaaagcgacc acccaacttt
      301 ggctgataat agcgtataaa caatgcatac tttgtacgtt caaaatacaa tgcagtagat
      361 atatttatgc atattacata taatacatat cacataggaa gcaacaggcg cgttggactt
      421 ttaattttcg aggaccgcga atccttacat cacacccaat cccccacaag tgatccccca
      481 cacaccatag cttcaaaatg tttctactcc ttttttactc ttccagattt tctcggactc
      541 cgcgcatcgc cgtaccactt caaaacaccc aagcacagca tactaaattt cccctctttc
      601 ttcctctagg gtgtcgttaa ttacccgtac taaaggtttg gaaaagaaaa aagacaccgc
      661 ctcgtttctt tttcttcgtc gaaaaaggca ataaaaattt ttatcacgtt tctttttctt
      721 gaaaattttt ttttttgatt tttttctctt tcgatgacct cccattgata tttaagttaa
      781 taaacggtca tcaatttctc aagtttcagt ttcatttttc ttgttctatt acaacttttt
      841 ttacttcttg ctcattagaa agaaagcata gcaatctaat ctaagtttta attacaaaag
      901 atctatgagt aagggtgaag aacttttcac tggagttgtc ccaattcttg ttgaattaga
      961 tggtgatgtt aatgggcaca aattttctgt cagtggagag ggtgaaggtg atgcaacata
     1021 cggaaaactt acccttaaat ttatttgcac tactggaaaa ctacctgttc catggccaac
     1081 acttgtcact actttcactt atggtgttca atgcttttca agatacccag atcatatgaa
     1141 acggcatgac tttttcaaga gtgccatgcc cgaaggttat gtacaggaaa gaactatatt
     1201 tttcaaagat gacgggaact acaagacacg tgctgaagtc aagtttgaag gtgataccct
     1261 tgttaataga atcgagttaa aaggtattga ttttaaagaa gatggaaaca ttcttggaca
     1321 caaattggaa tacaactata actcacacaa tgtatacatc atggcagaca aacaaaagaa
     1381 tggaatcaaa gttaacttca aaattagaca caacattgaa gatggaagcg ttcaactagc
     1441 agaccattat caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca
     1501 ttacctgtcc acacaatctg ccctttcgaa agatcccaac gaaaagagag atcacatggt
     1561 ccttcttgag tttgtaacag ctgctgggat tacacatggc atggatgaac tatacaaagg
     1621 ttctgaaaac ttgtacttcc aatctggtgc ttgtaatgat tcttctgatc cattggttgt
     1681 tgctgcttct attattggta ttttgcattt gattttgtgg attttggata gattgggatc
     1741 catggcttgt aaaaattggt tttcttcttt gtctcatttt gttatacatt tgtaactcga
     1801 gtggcagtgc ttttaactaa gaattattag tcttttctgc ttattttttc atcatagttt
     1861 agaacacttt atattaacga atagtttatg aatctattta ggtttaaaaa ttgatacagt
     1921 tttataagtt actttttcaa agactcgtgc tgtctattgc ataatgcact ggaaggggaa
     1981 aaaaaaggtg cacacgcgtg gctttttctt gaatttgcag tttgaaaaat gctgccaatg
     2041 agacgacggg gtcatcacgg ctcatcatgc gccaaacaaa tgtgtgcaat acacgctcgg
     2101 atgactgcat gatgaccgca ctgactgggg acagcagatc cacctaagcc tgtgagagaa
     2161 gcagacaccc gacagatcaa ggcagttaac tagtgcactg cagtacagtt tagcttgcct
     2221 cgtccccgcc gggtcacccg gccagcgaca tggaggccca gaataccctc cttgacagtc
     2281 ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag cccatacatc
     2341 cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga agcaaaaatt
     2401 acggctcctc gctccagacc tgcgagcagg gaaacgctcc cctcacagac gcgttgaatt
     2461 gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca ctgaggttct
     2521 tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc acatccgaac
     2581 ataaacaaaa atgggtaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
     2641 cgaaaagttc gacagcgtgt ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
     2701 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
     2761 tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga
     2821 agtgcttgac attggggaat ttagcgagag cctgacctat tgcatctccc gccgtgcaca
     2881 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc aaccggtcgc
     2941 ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt
     3001 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
     3061 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
     3121 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     3181 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
     3241 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag
     3301 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
     3361 tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
     3421 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
     3481 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt
     3541 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg
     3601 tccgagggca aaggaataaa gtaactgaca ataaaaagat tcttgttttc aagaacttgt
     3661 catttgtata gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt
     3721 atattttttt tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa
     3781 tatcatgcgt caatcgtatg tgaatgctgg tcgctatact ggagtaatgg aaggtcggga
     3841 tgagcatata caagcactaa gaagaacaat acagaactct acacggtatt attgtgctac
     3901 aagctcgagt aaaaccgagt gttttgacga tactaacgtt gttaagaaag taacttgtta
     3961 tcaaactcat taccaacttg tgattaattg gtgaataata tgataattgt cgaaattcca
     4021 ttgttggtaa agcctataat attatgtata cagattatac tagaaattct ctcgagaata
     4081 taagaatccc caaaattgaa tcggtatttc tacatactaa tattaccatt acttctcctt
     4141 tcgttttata tgtttcattc ctattacatt atcgatcttt gcatttcagc ttccattata
     4201 tttgatgtct gttttatgtc cccacgttac accgcatgtg acagtatact agtaacatga
     4261 gtgctaccga atagatgaca ttttagactt tcattccaac aacttggttg acagaatgtt
     4321 acgtacccta tatctaatct atatgaggcc tgaatctaac tgaaaggtgg aatttcagta
     4381 atttatcaag ctttaataag tttgggtagt ttaactgtgc aaaaaggtat ttaccttaca
     4441 tactgaatct tgtctgtttg gtagcggctg ctttatccga gcggccgcga ttatcaaaaa
     4501 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4561 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4621 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4681 gggagggctt accatctggc cccagtgctg caatgatacc gcgggaccca cgctcaccgg
     4741 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4801 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4861 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4921 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4981 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5041 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5101 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5161 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5221 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5281 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5341 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5401 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5461 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5521 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgtcatga
     5581 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5641 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5701 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5761 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     5821 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5881 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5941 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6001 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6061 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6121 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6181 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6241 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6301 tctttcctgc gttatcccct gattctgtgg ataaccgtgc ggccgccaat gccaggcgcc
     6361 tttatatcat ataattaaga cacaaaagga taaaacaaag gtgttaacta ttctgcatac
     6421 tcactatcgt aaactgtcct gcaaatcgtg taaatatgta tttcattttt tttgcagtga
     6481 aaaaaggcat gtaaaatacc gcatcaagta actctactcc gcctgtggtt tcaagactaa
     6541 cggcttgaga caaaatggga agaaatgatt gcagaaaagc catatgtgta atagcaaaaa
     6601 gctggatagt gcttaccaga tgtttacctt aatttcttgg tgaattagag aagtacagaa
     6661 gttttactat taatcccacc atagaaattt gtataggaaa gtagtttatt ggagttattg
     6721 gatatactgt gtaaactatt tcttgaaatt gtaatcttaa gatgctcttc ttattctatt
     6781 aaaaatagaa aatgattttc atatttattt atttatttat attttggcat tactcttcat
     6841 catttttttc cctctaagaa gcttcctttc tttttataag gataacaaaa ccaaaaggaa
     6901 tattgggtca gatgaatgga cgcgaatgca agacagaagt ccaaatcacg tcaagacaaa
     6961 gaaagaaaga aagaaaaact aacacattaa tgtagtttta aaatttcaaa tccgaacaac
     7021 agagcatagg gtttcgcaaa
//
