LOCUS       sequence-311466-        8624 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cytosolic labile heme reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8624)
  AUTHORS   Hanna DA, Harvey RM, Martinez-Guzman O, Yuan X, Chandrasekharan B, 
            Raju G, Outten FW, Hamza I, Reddi AR
  TITLE     Heme dynamics and trafficking factors revealed by genetically 
            encoded fluorescent heme sensors.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Jul 5;113(27):7539-44. doi: 
            10.1073/pnas.1523802113. Epub 2016 May 31.
  PUBMED    27247412
REFERENCE   2  (bases 1 to 8624)
  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"; doi: 
            "10.1073/pnas.1523802113"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2016-07-5- 5"; volume: "113"; issue: "27"; pages: "7539-44"
COMMENT     Sequence Label: yHS1-M7A
FEATURES             Location/Qualifiers
     source          1..8624
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     6..23
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    140..161
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(176..181,182..199,200..206)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    214..230
                     /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     219..241
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     238..254
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     238..254
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     273..293
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        275..293
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        318..961
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     855..878
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     complement(2751..2767)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2752..2768)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      2770..3017
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2786..2804)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(3035..3054)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3036..3054)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3064..3081)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3064..3080)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3073..3095)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3221..3676
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3308..3327)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3518..3539
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3977..4384
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             4385..5479
                     /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"
     primer_bind     complement(5973..5992)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6092..6114
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6152..6170)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    6212..6715
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6752..6856
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6857..6925,6926..7717)
                     /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(7075..7094)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7888..8476
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8377..8396
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
       61 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
      121 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
      181 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
      241 gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa
      301 agctggagct cagtttatca ttatcaatac tcgccatttc aaagaatacg taaataatta
      361 atagtagtga tttccctaac tttatttagt caaaaaatta gccttttaat tctgctgtaa
      421 cccgtacatg cccaaaatag ggggcgggtt acacagaata tataacatcg taggtgtctg
      481 ggtgaacagt ttattcctgg catccactaa atataatgga gcccgctttt taagctggca
      541 tccagaaaaa aaaagaatcc cagcaccaaa atattgtttt cttcaccaac catcagttca
      601 taggtccatt ctcttagcgc aactacagag aacaggggca caaacaggca aaaaacgggc
      661 acaacctcaa tggagtgatg caacctgcct ggagtaaatg atgacacaag gcaattgacc
      721 cacgcatgta tctatctcat tttcttacac cttctattac cttctgctct ctctgatttg
      781 gaaaaagctg aaaaaaaagg ttgaaaccag ttccctgaaa ttattcccct acttgactaa
      841 taagtatata aagacggtag gtattgattg taattctgta aatctatttc ttaaacttct
      901 taaattctac ttttatagtt agtctttttt ttagttttaa aacaccagaa cttagtttcg
      961 acggattcta gaactagtca tatgcacatg gtatcggaac tgatcaagga aaacatgcac
     1021 atgaagctgt atatggaagg cacggtcaac aaccaccact ttaaatgcac ctcagaaggc
     1081 gaaggtaaac cttatgaagg tacccaaact atgagaatta aggctgtcga aggtggccca
     1141 ctgccgtttg ccttcgatat cttagcgact tcttttatgt acggctcaaa gacattcatc
     1201 aaccatacgc aaggtatccc agatttcttt aaacagtcct ttccggaagg cttcacctgg
     1261 gaacgcgtta ccacttatga agacggtggc gtgctgacag ctacgcaaga tacttcgtta
     1321 caggacggtt gtttgattta caacgtcaaa atccgtggcg taaattttcc tagcaacggt
     1381 ccagtcatgc agaaaaagac attaggctgg gaagctagta ccgaaacttt gtatccggca
     1441 gatggtggcc tggaaggtcg cgctgacatg gcactgaaat tagtaggtgg cggtcacttg
     1501 atttgcaatc tgaagacaac gtaccgtagt aaaaagccgg ccaaaaactt aaagatgcct
     1561 ggtgtttatt acgtggatcg tagattggaa agaatcaaag aagcggacaa ggaaacgtat
     1621 gtggaacagc acgaagtcgc cgtcgcacgc tactgtgatt taccgtccaa gttaggtcat
     1681 agaggtggat ccatggtgtc caaaggtgaa gaactgttta ctggtgtagt gccgatttta
     1741 gttgaattag acggtgatgt gaatggtcat aagtttagcg tgtcgggtga aggcgaaggt
     1801 gatgctacct ttggtggctc tgcagatctg gaagacaacg cggaaactct gaacgataac
     1861 ttgaaggtta tcgaaaaggc cgataatgct gcacaagtga aagacgcgtt aacaaagatg
     1921 cgcgccgcgg ctttggatgc tcagaaagca acgccaccga aactggagga taagtctcct
     1981 gactcaccag aaatgaaaga ttttcgtcat ggcttcgaca ttttagtggg tcaaatcgat
     2041 gacgccttga aactggcgaa cgagggtaaa gtcaaggaag ctcaagcagc cgcggaacag
     2101 ctgaagacca ctcgtaatgc atatcatcag aaatacagag gtggcaaact gaccttgaag
     2161 tttatctgta caacgggtaa actgcctgtt ccatggccga ctttggtgac cactctgggc
     2221 tatggtgtac aatgcttctc acgttaccca gatcatatga aacagcacga ctttttcaag
     2281 agcgccatgc cggaaggcta cgttcaagaa cgcactattt tctttaagga tgacggtaac
     2341 tacaagacac gtgcggaagt caaatttgaa ggcgatacgc tggtaaacag aatcgaattg
     2401 aagggtatcg atttcaagga agacggcaac atcttgggtc ataaattgga atacaactac
     2461 aactcccaca acgtctacat catggctgat aagcaaaaga atggcatcaa ggtaaacttc
     2521 aagatcagac ataacatcga agatggttct gttcagctgg ctgaccacta tcaacagaat
     2581 acacctattg gcgatggtcc ggtgctgtta cctgataacc attacttgag cacgcagagt
     2641 gcactgtcca aagatccaaa tgaaaagcgc gaccacatgg tcttgttgga atttgtcacc
     2701 gctgctggta tcactttggg tatggatgaa ctgtataaat gaaagcttat cgataccgtc
     2761 gacctcgagt catgtaatta gttatgtcac gcttacattc acgccctccc cccacatccg
     2821 ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt tattttttta
     2881 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca
     2941 gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt ttgggacgct
     3001 cgaaggcttt aatttgcggc cggtacccaa ttcgccctat agtgagtcgt attacgcgcg
     3061 ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     3121 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     3181 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca
     3241 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3301 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3361 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3421 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3481 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3541 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3601 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     3661 ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3721 accgcatatc gaccggtcga ggagaacttc tagtatatct acatacctaa tattattgcc
     3781 ttattaaaaa tggaatccca acaattacat caaaatccac attctcttca aaatcaattg
     3841 tcctgtactt ccttgttcat gtgtgttcaa aaacgttata tttataggat aattatactc
     3901 tatttctcaa caagtaattg gttgtttggc cgagcggtct aaggcgcctg attcaagaaa
     3961 tatcttgacc gcagttaact gtgggaatac tcaggtatcg taagatgcaa gagttcgaat
     4021 ctcttagcaa ccattatttt tttcctcaac ataacgagaa cacacagggg cgctatcgca
     4081 cagaatcaaa ttcgatgact ggaaattttt tgttaatttc agaggtcgcc tgacgcatat
     4141 acctttttca actgaaaaat tgggagaaaa aggaaaggtg agagcgccgg aaccggcttt
     4201 tcatatagaa tagagaagcg ttcatgacta aatgcttgca tcacaatact tgaagttgac
     4261 aatattattt aaggacctat tgttttttcc aataggtggt tagcaatcgt cttactttct
     4321 aacttttctt accttttaca tttcagcaat atatatatat atatttcaag gatataccat
     4381 tctaatgtct gcccctaaga agatcgtcgt tttgccaggt gaccacgttg gtcaagaaat
     4441 cacagccgaa gccattaagg ttcttaaagc tatttctgat gttcgttcca atgtcaagtt
     4501 cgatttcgaa aatcatttaa ttggtggtgc tgctatcgat gctacaggtg ttccacttcc
     4561 agatgaggcg ctggaagcct ccaagaaggc tgatgccgtt ttgttaggtg ctgtgggtgg
     4621 tcctaaatgg ggtaccggta gtgttagacc tgaacaaggt ttactaaaaa tccgtaaaga
     4681 acttcaattg tacgccaact taagaccatg taactttgca tccgactctc ttttagactt
     4741 atctccaatc aagccacaat ttgctaaagg tactgacttc gttgttgtca gagaattagt
     4801 gggaggtatt tactttggta agagaaagga agacgatggt gatggtgtcg cttgggatag
     4861 tgaacaatac accgttccag aagtgcaaag aatcacaaga atggccgctt tcatggccct
     4921 acaacatgag ccaccattgc ctatttggtc cttggataaa gctaatgttt tggcctcttc
     4981 aagattatgg agaaaaactg tggaggaaac catcaagaac gaattcccta cattgaaggt
     5041 tcaacatcaa ttgattgatt ctgccgccat gatcctagtt aagaacccaa cccacctaaa
     5101 tggtattata atcaccagca acatgtttgg tgatatcatc tccgatgaag cctccgttat
     5161 cccaggttcc ttgggtttgt tgccatctgc gtccttggcc tctttgccag acaagaacac
     5221 cgcatttggt ttgtacgaac catgccacgg ttctgctcca gatttgccaa agaataaggt
     5281 caaccctatc gccactatct tgtctgctgc aatgatgttg aaattgtcat tgaacttgcc
     5341 tgaagaaggt aaggccattg aagatgcagt taaaaaggtt ttggatgcag gtatcagaac
     5401 tggtgattta ggtggttcca acagtaccac cgaagtcggt gatgctgtcg ccgaagaagt
     5461 taagaaaatc cttgcttaaa aagattctct ttttttatga tatttgtaca taaactttat
     5521 aaatgaaatt cataatagaa acgacacgaa attacaaaat ggaatatgtt catagggtag
     5581 acgaaactat atacgcaatc tacatacatt tatcaagaag gagaaaaagg aggatgtaaa
     5641 ggaatacagg taagcaaatt gatactaatg gctcaacgtg ataaggaaaa agaattgcac
     5701 tttaacatta atattgacaa ggaggagggc accacacaaa aagttaggtg taacagaaaa
     5761 tcatgaaact atgattccta atttatatat tggaggattt tctctaaaaa aaaaaaaata
     5821 caacaaataa aaaacactca atgacctgac catttgatgg agtttaagtc aataccttct
     5881 tgaaccattt cccataatgg tgaaagttcc ctcaagaatt ttactctgtc agaaacggcc
     5941 ttaacgacgt agtcgatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     6001 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     6061 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     6121 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     6181 aatgtcatga taataatggt ttcttaggac ggatcgcttg cctgtaactt acacgcgcct
     6241 cgtatctttt aatgatggaa taatttggga atttactctg tgtttattta tttttatgtt
     6301 ttgtatttgg attttagaaa gtaaataaag aaggtagaag agttacggaa tgaagaaaaa
     6361 aaaataaaca aaggtttaaa aaatttcaac aaaaagcgta ctttacatat atatttatta
     6421 gacaagaaaa gcagattaaa tagatataca ttcgattaac gataagtaaa atgtaaaatc
     6481 acaggatttt cgtgtgtggt cttctacaca gacaagatga aacaattcgg cattaatacc
     6541 tgagagcagg aagagcaaga taaaaggtag tatttgttgg cgatccccct agagtctttt
     6601 acatcttcgg aaaacaaaaa ctattttttc tttaatttct ttttttactt tctattttta
     6661 atttatatat ttatattaaa aaatttaaat tataattatt tttatagcac gtgatgaaaa
     6721 ggacccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     6781 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     6841 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     6901 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     6961 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     7021 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     7081 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     7141 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     7201 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     7261 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     7321 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     7381 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     7441 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     7501 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     7561 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     7621 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     7681 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     7741 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     7801 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     7861 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     7921 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     7981 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     8041 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     8101 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     8161 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     8221 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     8281 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     8341 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     8401 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     8461 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     8521 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     8581 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accg
//
