LOCUS       sequence-361502-        8684 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mitochondrial labile heme reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8684)
  AUTHORS   Martinez-Guzman O, Willoughby MM, Saini A, Dietz JV, Bohovych I, 
            Medlock AE, Khalimonchuk O, Reddi AR
  TITLE     Mitochondrial-nuclear heme trafficking in budding yeast is regulated
            by GTPases that control mitochondrial dynamics and ER contact sites.
  JOURNAL   J Cell Sci. 2020 May 20;133(10). pii: jcs.237917. doi: 
            10.1242/jcs.237917.
  PUBMED    32265272
REFERENCE   2  (bases 1 to 8684)
  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: "J Cell 
            Sci."; date: "2020-05-20"; volume: "133(10). pii: jcs.237917. doi"; 
            pages: " 10.1242/jcs.237917"
COMMENT     Sequence Label: mito-yHS1
FEATURES             Location/Qualifiers
     source          1..8684
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     72..91
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(310..414)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    451..954
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     996..1014
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1052..1074)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1174..1193
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(1687..2781)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(2782..3189)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(3490..3945)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3627..3648)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3839..3858
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4071..4093
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4085..4102
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4086..4102
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4112..4131
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4112..4130
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(4149..4396)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     4362..4380
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(6182..6247)
                     /codon_start=1
                     /product="mitochondrial presequence of yeast cytochrome c 
                     oxidase subunit IV (Hurt et al., 1984)"
                     /label=pre-CoxIV
                     /translation="MLSLRQSIRFFKPATRTLCSSR"
     promoter        complement(6265..6908)
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     complement(6348..6371)
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     complement(6933..6953)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6933..6951)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6972..6988)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6972..6988)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6985..7007)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6996..7012
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(7020..7026,7027..7044,7045..7050))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7065..7086
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7203..7220)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7374..7962)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7454..7473)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8133..8684,1..240,241..309))
                     /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"
ORIGIN
        1 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
       61 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
      121 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
      181 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
      241 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
      301 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
      361 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
      421 tccccgaaaa gtgccacctg ggtccttttc atcacgtgct ataaaaataa ttataattta
      481 aattttttaa tataaatata taaattaaaa atagaaagta aaaaaagaaa ttaaagaaaa
      541 aatagttttt gttttccgaa gatgtaaaag actctagggg gatcgccaac aaatactacc
      601 ttttatcttg ctcttcctgc tctcaggtat taatgccgaa ttgtttcatc ttgtctgtgt
      661 agaagaccac acacgaaaat cctgtgattt tacattttac ttatcgttaa tcgaatgtat
      721 atctatttaa tctgcttttc ttgtctaata aatatatatg taaagtacgc tttttgttga
      781 aattttttaa acctttgttt attttttttt cttcattccg taactcttct accttcttta
      841 tttactttct aaaatccaaa tacaaaacat aaaaataaat aaacacagag taaattccca
      901 aattattcca tcattaaaag atacgaggcg cgtgtaagtt acaggcaagc gatccgtcct
      961 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     1021 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     1081 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     1141 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     1201 tgcaccatat cgactacgtc gttaaggccg tttctgacag agtaaaattc ttgagggaac
     1261 tttcaccatt atgggaaatg gttcaagaag gtattgactt aaactccatc aaatggtcag
     1321 gtcattgagt gttttttatt tgttgtattt tttttttttt agagaaaatc ctccaatata
     1381 taaattagga atcatagttt catgattttc tgttacacct aactttttgt gtggtgccct
     1441 cctccttgtc aatattaatg ttaaagtgca attctttttc cttatcacgt tgagccatta
     1501 gtatcaattt gcttacctgt attcctttac atcctccttt ttctccttct tgataaatgt
     1561 atgtagattg cgtatatagt ttcgtctacc ctatgaacat attccatttt gtaatttcgt
     1621 gtcgtttcta ttatgaattt catttataaa gtttatgtac aaatatcata aaaaaagaga
     1681 atctttttaa gcaaggattt tcttaacttc ttcggcgaca gcatcaccga cttcggtggt
     1741 actgttggaa ccacctaaat caccagttct gatacctgca tccaaaacct ttttaactgc
     1801 atcttcaatg gccttacctt cttcaggcaa gttcaatgac aatttcaaca tcattgcagc
     1861 agacaagata gtggcgatag ggttgacctt attctttggc aaatctggag cagaaccgtg
     1921 gcatggttcg tacaaaccaa atgcggtgtt cttgtctggc aaagaggcca aggacgcaga
     1981 tggcaacaaa cccaaggaac ctgggataac ggaggcttca tcggagatga tatcaccaaa
     2041 catgttgctg gtgattataa taccatttag gtgggttggg ttcttaacta ggatcatggc
     2101 ggcagaatca atcaattgat gttgaacctt caatgtaggg aattcgttct tgatggtttc
     2161 ctccacagtt tttctccata atcttgaaga ggccaaaaca ttagctttat ccaaggacca
     2221 aataggcaat ggtggctcat gttgtagggc catgaaagcg gccattcttg tgattctttg
     2281 cacttctgga acggtgtatt gttcactatc ccaagcgaca ccatcaccat cgtcttcctt
     2341 tctcttacca aagtaaatac ctcccactaa ttctctgaca acaacgaagt cagtaccttt
     2401 agcaaattgt ggcttgattg gagataagtc taaaagagag tcggatgcaa agttacatgg
     2461 tcttaagttg gcgtacaatt gaagttcttt acggattttt agtaaacctt gttcaggtct
     2521 aacactaccg gtaccccatt taggaccacc cacagcacct aacaaaacgg catcagcctt
     2581 cttggaggct tccagcgcct catctggaag tggaacacct gtagcatcga tagcagcacc
     2641 accaattaaa tgattttcga aatcgaactt gacattggaa cgaacatcag aaatagcttt
     2701 aagaacctta atggcttcgg ctgtgatttc ttgaccaacg tggtcacctg gcaaaacgac
     2761 gatcttctta ggggcagaca ttagaatggt atatccttga aatatatata tatatattgc
     2821 tgaaatgtaa aaggtaagaa aagttagaaa gtaagacgat tgctaaccac ctattggaaa
     2881 aaacaatagg tccttaaata atattgtcaa cttcaagtat tgtgatgcaa gcatttagtc
     2941 atgaacgctt ctctattcta tatgaaaagc cggttccggc gctctcacct ttcctttttc
     3001 tcccaatttt tcagttgaaa aaggtatatg cgtcaggcga cctctgaaat taacaaaaaa
     3061 tttccagtca tcgaatttga ttctgtgcga tagcgcccct gtgtgttctc gttatgttga
     3121 ggaaaaaaat aatggttgct aagagattcg aactcttgca tcttacgata cctgagtatt
     3181 cccacagtta actgcggtca agatatttct tgaatcaggc gccttagacc gctcggccaa
     3241 acaaccaatt acttgttgag aaatagagta taattatcct ataaatataa cgtttttgaa
     3301 cacacatgaa caaggaagta caggacaatt gattttgaag agaatgtgga ttttgatgta
     3361 attgttggga ttccattttt aataaggcaa taatattagg tatgtagata tactagaagt
     3421 tctcctcgac cggtcgatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     3481 cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     3541 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     3601 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     3661 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     3721 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     3781 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     3841 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     3901 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt cgccattcag
     3961 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     4021 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     4081 acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg
     4141 taccggccgc aaattaaagc cttcgagcgt cccaaaacct tctcaagcaa ggttttcagt
     4201 ataatgttac atgcgtacac gcgtctgtac agaaaaaaaa gaaaaatttg aaatataaat
     4261 aacgttctta atactaacat aactataaaa aaataaatag ggacctagac ttcaggttgt
     4321 ctaactcctt ccttttcggt tagagcggat gtggggggag ggcgtgaatg taagcgtgac
     4381 ataactaatt acatgactcg agtcatttat acagttcatc catacccaaa gtgataccag
     4441 cagcggtgac aaattccaac aagaccatgt ggtcgcgctt ttcatttgga tctttggaca
     4501 gtgcactctg cgtgctcaag taatggttat caggtaacag caccggacca tcgccaatag
     4561 gtgtattctg ttgatagtgg tcagccagct gaacagaacc atcttcgatg ttatgtctga
     4621 tcttgaagtt taccttgatg ccattctttt gcttatcagc catgatgtag acgttgtggg
     4681 agttgtagtt gtattccaat ttatgaccca agatgttgcc gtcttccttg aaatcgatac
     4741 ccttcaattc gattctgttt accagcgtat cgccttcaaa tttgacttcc gcacgtgtct
     4801 tgtagttacc gtcatcctta aagaaaatag tgcgttcttg aacgtagcct tccggcatgg
     4861 cgctcttgaa aaagtcgtgc tgtttcatat gatctgggta acgtgagaag cattgtacac
     4921 catagcccag agtggtcacc aaagtcggcc atggaacagg cagtttaccc gttgtacaga
     4981 taaacttcaa ggtcagtttg ccacctctgt atttctgatg atatgcatta cgagtggtct
     5041 tcagctgttc cgcggctgct tgagcttcct tgactttacc ctcgttcgcc agtttcaagg
     5101 cgtcatcgat ttgacccact aaaatgtcga agccatgacg aaaatctttc atttctggtg
     5161 agtcaggaga cttatcctcc agtttcggtg gcgttgcttt ctgagcatcc aaagccgcgg
     5221 cgcgcatctt tgttaacgcg tctttcactt gtgcagcatt atcggccttt tcgataacct
     5281 tcaagttatc gttcagagtt tccatgttgt cttccagatc tgcagagcca ccaaaggtag
     5341 catcaccttc gccttcaccc gacacgctaa acttatgacc attcacatca ccgtctaatt
     5401 caactaaaat cggcactaca ccagtaaaca gttcttcacc tttggacacc atggatccac
     5461 ctctatgacc taacttggac ggtaaatcac agtagcgtgc gacggcgact tcgtgctgtt
     5521 ccacatacgt ttccttgtcc gcttctttga ttctttccaa tctacgatcc acgtaataaa
     5581 caccaggcat ctttaagttt ttggccggct ttttactacg gtacgttgtc ttcagattgc
     5641 aaatcaagtg accgccacct actaatttca gtgccatgtc agcgcgacct tccaggccac
     5701 catctgccgg atacaaagtt tcggtactag cttcccagcc taatgtcttt ttctgcatga
     5761 ctggaccgtt gctaggaaaa tttacgccac ggattttgac gttgtaaatc aaacaaccgt
     5821 cctgtaacga agtatcttgc gtagctgtca gcacgccacc gtcttcataa gtggtaacgc
     5881 gttcccaggt gaagccttcc ggaaaggact gtttaaagaa atctgggata ccttgcgtat
     5941 ggttgatgaa tgtctttgag ccgtacataa aagaagtcgc taagatatcg aaggcaaacg
     6001 gcagtgggcc accttcgaca gccttaattc tcatagtttg ggtaccttca taaggtttac
     6061 cttcgccttc tgaggtgcat ttaaagtggt ggttgttgac cgtgccttcc atatacagct
     6121 tcatgtgcat gttttccttg atcagttccg ataccatgtg catatgaagc ttaagcagat
     6181 atctagagct acacaaagtt cttgtggctg gcttgaaaaa tcttatagat tgacgtagtg
     6241 aaagcatact agttctagaa tccgtcgaaa ctaagttctg gtgttttaaa actaaaaaaa
     6301 agactaacta taaaagtaga atttaagaag tttaagaaat agatttacag aattacaatc
     6361 aatacctacc gtctttatat acttattagt caagtagggg aataatttca gggaactggt
     6421 ttcaaccttt tttttcagct ttttccaaat cagagagagc agaaggtaat agaaggtgta
     6481 agaaaatgag atagatacat gcgtgggtca attgccttgt gtcatcattt actccaggca
     6541 ggttgcatca ctccattgag gttgtgcccg ttttttgcct gtttgtgccc ctgttctctg
     6601 tagttgcgct aagagaatgg acctatgaac tgatggttgg tgaagaaaac aatattttgg
     6661 tgctgggatt cttttttttt ctggatgcca gcttaaaaag cgggctccat tatatttagt
     6721 ggatgccagg aataaactgt tcacccagac acctacgatg ttatatattc tgtgtaaccc
     6781 gccccctatt ttgggcatgt acgggttaca gcagaattaa aaggctaatt ttttgactaa
     6841 ataaagttag gaaaatcact actattaatt atttacgtat tctttgaaat ggcgagtatt
     6901 gataatgata aactgagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     6961 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     7021 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     7081 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     7141 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     7201 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     7261 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     7321 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     7381 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     7441 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     7501 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     7561 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     7621 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     7681 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     7741 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     7801 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     7861 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     7921 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7981 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     8041 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     8101 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     8161 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     8221 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     8281 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     8341 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     8401 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     8461 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     8521 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     8581 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     8641 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtga
//
