LOCUS       sequence-342886-        7132 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human ferrochelatase (FECH) in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7132)
  AUTHORS   Dietz JV, Willoughby MM, Piel RB 3rd, Ross TA, Bohovych I, Addis HG,
            Fox JL, Lanzilotta WN, Dailey HA, Wohlschlegel JA, Reddi AR, Medlock
            AE, Khalimonchuk O
  TITLE     Mitochondrial contact site and cristae organizing system (MICOS) 
            machinery supports heme biosynthesis by enabling optimal performance
            of ferrochelatase.
  JOURNAL   Redox Biol. 2021 Oct;46:102125. doi: 10.1016/j.redox.2021.102125. 
            Epub 2021 Sep 10.
  PUBMED    34517185
REFERENCE   2  (bases 1 to 7132)
  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: "Redox 
            Biol."; date: "2021-10"; volume: "46:102125. doi"; pages: " 
            10.1016/j.redox.2021.102125. Epub 2021 Sep 10"
COMMENT     Sequence Label: pRS426-HsFECH
FEATURES             Location/Qualifiers
     source          1..7132
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        194..409
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             410..1213
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1344..1799)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1481..1502)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1693..1712
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1925..1947
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1939..1956
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1940..1956
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1966..1985
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1966..1984
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2009..2025
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2010..2026
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3126..3143)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3530..3550)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3530..3548)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3569..3585)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3569..3585)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3582..3604)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3593..3609
                     /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(3617..3623,3624..3641,3642..3647))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3662..3683
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3800..3817)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3971..4559)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4051..4070)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4730..5521,5522..5590))
                     /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     5353..5372
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5591..5695)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5722..7064
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(6685..6732)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     7105..7123
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac cttttcaatt catcattttt tttttattct tttttttgat ttcggtttcc
      241 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
      301 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt
      361 cttaacccaa ctgcacagaa caaaaacctg caggaaacga agataaatca tgtcgaaagc
      421 tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat
      481 catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt
      541 actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat
      601 cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta
      661 caatttttta ctcttcgaag acagaaaatt tgctgacatt ggtaatacag tcaaattgca
      721 gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt
      781 ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc
      841 tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata
      901 tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc
      961 tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt
     1021 gggtttagat gacaagggag acgcattggg tcaacagtat agaaccgtgg atgatgtggt
     1081 ctctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc
     1141 taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg
     1201 ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag
     1261 agcttcaatt taattatatc agttattacc ctatgcggtg tgaaataccg cacagatgcg
     1321 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     1381 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     1441 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     1501 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     1561 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     1621 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     1681 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     1741 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     1801 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     1861 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     1921 ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta
     1981 tagggcgaat tgggtaccgg gccccccctc gaggtcgacg gtatcgataa gcttcacagc
     2041 tgctggctgg tgaagaagga tttagtctcc ctgcagacag gattgacaca gagcggacag
     2101 ctcagggtca gctgcttgga acacagctcg tttgactgga tgtgtgaatg caccaagtcg
     2161 gccagggcct tagagaacaa tggatttcca ttaagagact cagctcttct gatgttttca
     2221 actccacact ccttggctaa aacttgagag tactcgatgt ccagctcata cagcgtttca
     2281 atatggtcac tggtaaatgc tatcggaacc aagaggatat tcttcctccc cctctcacaa
     2341 agccctttga tagattcgtc tgtttgagga cccaaccagg gcatcggacc aaccttggat
     2401 tgccacacca gtcggtaggg gttgcagtac tccagccttt ccatgacttt ttggacagtg
     2461 gcgcttacct cctgaggata tgggtcgcct ctgttgacca cagacatcgg cagtgagtga
     2521 gcagaaaaca gaatgaccac ctcgcttctc ttctcaagtg gaaaatggtc cagttccttt
     2581 agaatatgat ctgcaaagca ctggatgagg aggtgatgtg tgggccacct gtcaatagtg
     2641 ctccacttca tcgtgggctt ccgtcccact tgattatagt atctgtaaat ggcatttaag
     2701 ctgctgcctg tggtggagca gctgtactgt ggatactgtg tgaaagcaat agccctttct
     2761 aggccatctc tctccatctc ttcaattgct tcttctgtta aaggatggac gtaccgaaat
     2821 ccaatatagt atttgtgagg ggctgtgttg ggggacaatt catccagcag cttcaccatg
     2881 ccctctccct gcttggaagt ccatatcttg atgggggatc cgcctccaat cctgcggtac
     2941 tgctcttgaa tcttgggggt tcggcgtttg gcgatgaatg gtgccagctt attctgaata
     3001 ggaagtgtca tgaggtctcg gtccaagaag agtctcagaa ggaagtcgtg aacatctcca
     3061 agagtttcag ggcctcccat gtttagcatt aatattccag ttttcggctt cctcttctgc
     3121 ggttgatggt ggtgatggtg gtgagccatg gtgaatgtaa ccgaaaatga tcttgtaatg
     3181 gtcagttgtg atcttcttag gaaggaacct tgtgtacgga ttgttctgga aagcattctt
     3241 taaacgttct tttttttact gctaaatttg tccaaatagc aggtatgtgt taaatatttc
     3301 tgtacttctt gtaaaccgac tcaagtggaa cacgagaatt tacgcttgtt tggtgtcaag
     3361 ccaagagtga ccttgaccga ccaaaagtat tccttcgcgt cctccttctt caatcccata
     3421 tcccctacga aaaaaagcag catgagaaaa cgaagggcag caactccttc gtcatataag
     3481 gcttacggtg accgcggccg ccaccgcggt ggagctccag cttttgttcc ctttagtgag
     3541 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     3601 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     3661 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3721 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3781 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3841 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3901 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3961 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4021 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4081 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4141 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4201 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4261 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4321 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4381 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4441 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4501 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4561 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4621 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4681 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4741 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4801 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4861 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4921 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4981 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5041 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5101 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5161 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5221 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5281 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5341 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5401 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5461 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5521 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5581 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5641 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5701 ttccccgaaa agtgccacct gaacgaagca tctgtgcttc attttgtaga acaaaaatgc
     5761 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     5821 tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa
     5881 aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac
     5941 agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct
     6001 acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc
     6061 tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag
     6121 gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact
     6181 tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc
     6241 ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt
     6301 tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac
     6361 tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt
     6421 actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg
     6481 agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca
     6541 cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata
     6601 ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcttcagagc
     6661 gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagagaata ggaacttcgg
     6721 aataggaact tcaaagcgtt tccgaaaacg agcgcttccg aaaatgcaac gcgagctgcg
     6781 cacatacagc tcactgttca cgtcgcacct atatctgcgt gttgcctgta tatatatata
     6841 catgagaaga acggcatagt gcgtgtttat gcttaaatgc gtacttatat gcgtctattt
     6901 atgtaggatg aaaggtagtc tagtacctcc tgtgatatta tcccattcca tgcggggtat
     6961 cgtatgcttc cttcagcact accctttagc tgttctatat gctgccactc ctcaattgga
     7021 ttagtctcat ccttcaatgc tatcatttcc tttgatattg gatcatacta agaaaccatt
     7081 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tc
//
