LOCUS       sequence-361504-        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-M7A
FEATURES             Location/Qualifiers
     source          1..8684
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(388..407)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     507..529
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(567..585)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    627..1130
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        1167..1271
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1272..1340,1341..2132)
                     /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     complement(1490..1509)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2303..2891
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2792..2811
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3045..3062
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3179..3200
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3215..3220,3221..3238,3239..3245)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3253..3269
                     /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)."
     primer_bind     3258..3280
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3277..3293
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3277..3293
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3312..3332
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3314..3332
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3357..4000
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     3894..3917
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             4018..4083
                     /codon_start=1
                     /product="mitochondrial presequence of yeast cytochrome c 
                     oxidase subunit IV (Hurt et al., 1984)"
                     /label=pre-CoxIV
                     /translation="MLSLRQSIRFFKPATRTLCSSR"
     terminator      5869..6116
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5885..5903)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(6134..6153)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6135..6153)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6163..6180)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6163..6179)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6172..6194)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6320..6775
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6407..6426)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6617..6638
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7076..7483
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             7484..8578
                     /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"
ORIGIN
        1 actatatacg caatctacat acatttatca agaaggagaa aaaggaggat gtaaaggaat
       61 acaggtaagc aaattgatac taatggctca acgtgataag gaaaaagaat tgcactttaa
      121 cattaatatt gacaaggagg agggcaccac acaaaaagtt aggtgtaaca gaaaatcatg
      181 aaactatgat tcctaattta tatattggag gattttctct aaaaaaaaaa aaatacaaca
      241 aataaaaaac actcaatgac ctgaccattt gatggagttt aagtcaatac cttcttgaac
      301 catttcccat aatggtgaaa gttccctcaa gaattttact ctgtcagaaa cggccttaac
      361 gacgtagtcg atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
      421 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
      481 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
      541 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
      601 catgataata atggtttctt aggacggatc gcttgcctgt aacttacacg cgcctcgtat
      661 cttttaatga tggaataatt tgggaattta ctctgtgttt atttattttt atgttttgta
      721 tttggatttt agaaagtaaa taaagaaggt agaagagtta cggaatgaag aaaaaaaaat
      781 aaacaaaggt ttaaaaaatt tcaacaaaaa gcgtacttta catatatatt tattagacaa
      841 gaaaagcaga ttaaatagat atacattcga ttaacgataa gtaaaatgta aaatcacagg
      901 attttcgtgt gtggtcttct acacagacaa gatgaaacaa ttcggcatta atacctgaga
      961 gcaggaagag caagataaaa ggtagtattt gttggcgatc cccctagagt cttttacatc
     1021 ttcggaaaac aaaaactatt ttttctttaa tttctttttt tactttctat ttttaattta
     1081 tatatttata ttaaaaaatt taaattataa ttatttttat agcacgtgat gaaaaggacc
     1141 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     1201 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     1261 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     1321 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     1381 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     1441 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     1501 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     1561 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     1621 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     1681 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     1741 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     1801 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     1861 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     1921 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     1981 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     2041 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     2101 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     2161 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     2221 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     2281 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2341 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2401 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     2461 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2521 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2581 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2641 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2701 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2761 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     2821 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     2881 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     2941 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3001 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3061 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     3121 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     3181 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     3241 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     3301 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctc agtttatcat
     3361 tatcaatact cgccatttca aagaatacgt aaataattaa tagtagtgat tttcctaact
     3421 ttatttagtc aaaaaattag ccttttaatt ctgctgtaac ccgtacatgc ccaaaatagg
     3481 gggcgggtta cacagaatat ataacatcgt aggtgtctgg gtgaacagtt tattcctggc
     3541 atccactaaa tataatggag cccgcttttt aagctggcat ccagaaaaaa aaagaatccc
     3601 agcaccaaaa tattgttttc ttcaccaacc atcagttcat aggtccattc tcttagcgca
     3661 actacagaga acaggggcac aaacaggcaa aaaacgggca caacctcaat ggagtgatgc
     3721 aacctgcctg gagtaaatga tgacacaagg caattgaccc acgcatgtat ctatctcatt
     3781 ttcttacacc ttctattacc ttctgctctc tctgatttgg aaaaagctga aaaaaaaggt
     3841 tgaaaccagt tccctgaaat tattccccta cttgactaat aagtatataa agacggtagg
     3901 tattgattgt aattctgtaa atctatttct taaacttctt aaattctact tttatagtta
     3961 gtcttttttt tagttttaaa acaccagaac ttagtttcga cggattctag aactagtatg
     4021 ctttcactac gtcaatctat aagatttttc aagccagcca caagaacttt gtgtagctct
     4081 agatatctgc ttaagcttca tatgcacatg gtatcggaac tgatcaagga aaacatgcac
     4141 atgaagctgt atatggaagg cacggtcaac aaccaccact ttaaatgcac ctcagaaggc
     4201 gaaggtaaac cttatgaagg tacccaaact atgagaatta aggctgtcga aggtggccca
     4261 ctgccgtttg ccttcgatat cttagcgact tcttttatgt acggctcaaa gacattcatc
     4321 aaccatacgc aaggtatccc agatttcttt aaacagtcct ttccggaagg cttcacctgg
     4381 gaacgcgtta ccacttatga agacggtggc gtgctgacag ctacgcaaga tacttcgtta
     4441 caggacggtt gtttgattta caacgtcaaa atccgtggcg taaattttcc tagcaacggt
     4501 ccagtcatgc agaaaaagac attaggctgg gaagctagta ccgaaacttt gtatccggca
     4561 gatggtggcc tggaaggtcg cgctgacatg gcactgaaat tagtaggtgg cggtcacttg
     4621 atttgcaatc tgaagacaac gtaccgtagt aaaaagccgg ccaaaaactt aaagatgcct
     4681 ggtgtttatt acgtggatcg tagattggaa agaatcaaag aagcggacaa ggaaacgtat
     4741 gtggaacagc acgaagtcgc cgtcgcacgc tactgtgatt taccgtccaa gttaggtcat
     4801 agaggtggat ccatggtgtc caaaggtgaa gaactgttta ctggtgtagt gccgatttta
     4861 gttgaattag acggtgatgt gaatggtcat aagtttagcg tgtcgggtga aggcgaaggt
     4921 gatgctacct ttggtggctc tgcagatctg gaagacaacg cggaaactct gaacgataac
     4981 ttgaaggtta tcgaaaaggc cgataatgct gcacaagtga aagacgcgtt aacaaagatg
     5041 cgcgccgcgg ctttggatgc tcagaaagca acgccaccga aactggagga taagtctcct
     5101 gactcaccag aaatgaaaga ttttcgtcat ggcttcgaca ttttagtggg tcaaatcgat
     5161 gacgccttga aactggcgaa cgagggtaaa gtcaaggaag ctcaagcagc cgcggaacag
     5221 ctgaagacca ctcgtaatgc atatcatcag aaatacagag gtggcaaact gaccttgaag
     5281 tttatctgta caacgggtaa actgcctgtt ccatggccga ctttggtgac cactctgggc
     5341 tatggtgtac aatgcttctc acgttaccca gatcatatga aacagcacga ctttttcaag
     5401 agcgccatgc cggaaggcta cgttcaagaa cgcactattt tctttaagga tgacggtaac
     5461 tacaagacac gtgcggaagt caaatttgaa ggcgatacgc tggtaaacag aatcgaattg
     5521 aagggtatcg atttcaagga agacggcaac atcttgggtc ataaattgga atacaactac
     5581 aactcccaca acgtctacat catggctgat aagcaaaaga atggcatcaa ggtaaacttc
     5641 aagatcagac ataacatcga agatggttct gttcagctgg ctgaccacta tcaacagaat
     5701 acacctattg gcgatggtcc ggtgctgtta cctgataacc attacttgag cacgcagagt
     5761 gcactgtcca aagatccaaa tgaaaagcgc gaccacatgg tcttgttgga atttgtcacc
     5821 gctgctggta tcactttggg tatggatgaa ctgtataaat gactcgagtc atgtaattag
     5881 ttatgtcacg cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag
     5941 ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa
     6001 cgttatttat atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca
     6061 ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaaggcttta atttgcggcc
     6121 ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca
     6181 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     6241 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     6301 cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     6361 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     6421 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     6481 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     6541 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     6601 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     6661 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     6721 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga
     6781 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatcg accggtcgag
     6841 gagaacttct agtatatcta catacctaat attattgcct tattaaaaat ggaatcccaa
     6901 caattacatc aaaatccaca ttctcttcaa aatcaattgt cctgtacttc cttgttcatg
     6961 tgtgttcaaa aacgttatat ttataggata attatactct atttctcaac aagtaattgg
     7021 ttgtttggcc gagcggtcta aggcgcctga ttcaagaaat atcttgaccg cagttaactg
     7081 tgggaatact caggtatcgt aagatgcaag agttcgaatc tcttagcaac cattattttt
     7141 ttcctcaaca taacgagaac acacaggggc gctatcgcac agaatcaaat tcgatgactg
     7201 gaaatttttt gttaatttca gaggtcgcct gacgcatata cctttttcaa ctgaaaaatt
     7261 gggagaaaaa ggaaaggtga gagcgccgga accggctttt catatagaat agagaagcgt
     7321 tcatgactaa atgcttgcat cacaatactt gaagttgaca atattattta aggacctatt
     7381 gttttttcca ataggtggtt agcaatcgtc ttactttcta acttttctta ccttttacat
     7441 ttcagcaata tatatatata tatttcaagg atataccatt ctaatgtctg cccctaagaa
     7501 gatcgtcgtt ttgccaggtg accacgttgg tcaagaaatc acagccgaag ccattaaggt
     7561 tcttaaagct atttctgatg ttcgttccaa tgtcaagttc gatttcgaaa atcatttaat
     7621 tggtggtgct gctatcgatg ctacaggtgt tccacttcca gatgaggcgc tggaagcctc
     7681 caagaaggct gatgccgttt tgttaggtgc tgtgggtggt cctaaatggg gtaccggtag
     7741 tgttagacct gaacaaggtt tactaaaaat ccgtaaagaa cttcaattgt acgccaactt
     7801 aagaccatgt aactttgcat ccgactctct tttagactta tctccaatca agccacaatt
     7861 tgctaaaggt actgacttcg ttgttgtcag agaattagtg ggaggtattt actttggtaa
     7921 gagaaaggaa gacgatggtg atggtgtcgc ttgggatagt gaacaataca ccgttccaga
     7981 agtgcaaaga atcacaagaa tggccgcttt catggcccta caacatgagc caccattgcc
     8041 tatttggtcc ttggataaag ctaatgtttt ggcctcttca agattatgga gaaaaactgt
     8101 ggaggaaacc atcaagaacg aattccctac attgaaggtt caacatcaat tgattgattc
     8161 tgccgccatg atcctagtta agaacccaac ccacctaaat ggtattataa tcaccagcaa
     8221 catgtttggt gatatcatct ccgatgaagc ctccgttatc ccaggttcct tgggtttgtt
     8281 gccatctgcg tccttggcct ctttgccaga caagaacacc gcatttggtt tgtacgaacc
     8341 atgccacggt tctgctccag atttgccaaa gaataaggtc aaccctatcg ccactatctt
     8401 gtctgctgca atgatgttga aattgtcatt gaacttgcct gaagaaggta aggccattga
     8461 agatgcagtt aaaaaggttt tggatgcagg tatcagaact ggtgatttag gtggttccaa
     8521 cagtaccacc gaagtcggtg atgctgtcgc cgaagaagtt aagaaaatcc ttgcttaaaa
     8581 agattctctt tttttatgat atttgtacat aaactttata aatgaaattc ataatagaaa
     8641 cgacacgaaa ttacaaaatg gaatatgttc atagggtaga cgaa
//
