LOCUS       sequence-311475-        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,H102A
FEATURES             Location/Qualifiers
     source          1..8624
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    127..148
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(163..168,169..186,187..193)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    201..217
                     /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     206..228
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     225..241
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     225..241
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     260..280
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        262..280
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        305..948
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     842..865
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     complement(2738..2754)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2739..2755)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      2757..3004
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2773..2791)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(3022..3041)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3023..3041)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3051..3068)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3051..3067)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3060..3082)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3208..3663
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3295..3314)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3505..3526
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3964..4371
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             4372..5466
                     /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(5960..5979)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6079..6101
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6139..6157)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    6199..6702
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6739..6843
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6844..6912,6913..7704)
                     /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(7062..7081)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7875..8463
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8364..8383
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(8617..8624,1..10)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
       61 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
      121 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
      181 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
      241 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctcag
      301 tttatcatta tcaatactcg ccatttcaaa gaatacgtaa ataattaata gtagtgattt
      361 tcctaacttt atttagtcaa aaaattagcc ttttaattct gctgtaaccc gtacatgccc
      421 aaaatagggg gcgggttaca cagaatatat aacatcgtag gtgtctgggt gaacagttta
      481 ttcctggcat ccactaaata taatggagcc cgctttttaa gctggcatcc agaaaaaaaa
      541 agaatcccag caccaaaata ttgttttctt caccaaccat cagttcatag gtccattctc
      601 ttagcgcaac tacagagaac aggggcacaa acaggcaaaa aacgggcaca acctcaatgg
      661 agtgatgcaa cctgcctgga gtaaatgatg acacaaggca attgacccac gcatgtatct
      721 atctcatttt cttacacctt ctattacctt ctgctctctc tgatttggaa aaagctgaaa
      781 aaaaaggttg aaaccagttc cctgaaatta ttcccctact tgactaataa gtatataaag
      841 acggtaggta ttgattgtaa ttctgtaaat ctatttctta aacttcttaa attctacttt
      901 tatagttagt ctttttttta gttttaaaac accagaactt agtttcgacg gattctagaa
      961 ctagtcatat gcacatggta tcggaactga tcaaggaaaa catgcacatg aagctgtata
     1021 tggaaggcac ggtcaacaac caccacttta aatgcacctc agaaggcgaa ggtaaacctt
     1081 atgaaggtac ccaaactatg agaattaagg ctgtcgaagg tggcccactg ccgtttgcct
     1141 tcgatatctt agcgacttct tttatgtacg gctcaaagac attcatcaac catacgcaag
     1201 gtatcccaga tttctttaaa cagtcctttc cggaaggctt cacctgggaa cgcgttacca
     1261 cttatgaaga cggtggcgtg ctgacagcta cgcaagatac ttcgttacag gacggttgtt
     1321 tgatttacaa cgtcaaaatc cgtggcgtaa attttcctag caacggtcca gtcatgcaga
     1381 aaaagacatt aggctgggaa gctagtaccg aaactttgta tccggcagat ggtggcctgg
     1441 aaggtcgcgc tgacatggca ctgaaattag taggtggcgg tcacttgatt tgcaatctga
     1501 agacaacgta ccgtagtaaa aagccggcca aaaacttaaa gatgcctggt gtttattacg
     1561 tggatcgtag attggaaaga atcaaagaag cggacaagga aacgtatgtg gaacagcacg
     1621 aagtcgccgt cgcacgctac tgtgatttac cgtccaagtt aggtcataga ggtggatcca
     1681 tggtgtccaa aggtgaagaa ctgtttactg gtgtagtgcc gattttagtt gaattagacg
     1741 gtgatgtgaa tggtcataag tttagcgtgt cgggtgaagg cgaaggtgat gctacctttg
     1801 gtggctctgc agatctggaa gacaacgcgg aaactctgaa cgataacttg aaggttatcg
     1861 aaaaggccga taatgctgca caagtgaaag acgcgttaac aaagatgcgc gccgcggctt
     1921 tggatgctca gaaagcaacg ccaccgaaac tggaggataa gtctcctgac tcaccagaaa
     1981 tgaaagattt tcgtaatggc ttcgacattt tagtgggtca aatcgatgac gccttgaaac
     2041 tggcgaacga gggtaaagtc aaggaagctc aagcagccgc ggaacagctg aagaccactc
     2101 gtaatgcata tgctcagaaa tacagaggtg gcaaactgac cttgaagttt atctgtacaa
     2161 cgggtaaact gcctgttcca tggccgactt tggtgaccac tctgggctat ggtgtacaat
     2221 gcttctcacg ttacccagat catatgaaac agcacgactt tttcaagagc gccatgccgg
     2281 aaggctacgt tcaagaacgc actattttct ttaaggatga cggtaactac aagacacgtg
     2341 cggaagtcaa atttgaaggc gatacgctgg taaacagaat cgaattgaag ggtatcgatt
     2401 tcaaggaaga cggcaacatc ttgggtcata aattggaata caactacaac tcccacaacg
     2461 tctacatcat ggctgataag caaaagaatg gcatcaaggt aaacttcaag atcagacata
     2521 acatcgaaga tggttctgtt cagctggctg accactatca acagaataca cctattggcg
     2581 atggtccggt gctgttacct gataaccatt acttgagcac gcagagtgca ctgtccaaag
     2641 atccaaatga aaagcgcgac cacatggtct tgttggaatt tgtcaccgct gctggtatca
     2701 ctttgggtat ggatgaactg tataaatgaa agcttatcga taccgtcgac ctcgagtcat
     2761 gtaattagtt atgtcacgct tacattcacg ccctcccccc acatccgctc taaccgaaaa
     2821 ggaaggagtt agacaacctg aagtctaggt ccctatttat ttttttatag ttatgttagt
     2881 attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc
     2941 atgtaacatt atactgaaaa ccttgcttga gaaggttttg ggacgctcga aggctttaat
     3001 ttgcggccgg tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc
     3061 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     3121 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     3181 cagttgcgca gcctgaatgg cgaatggacg cgccctgtag cggcgcatta agcgcggcgg
     3241 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3301 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3361 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3421 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3481 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3541 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3601 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     3661 tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatcgac
     3721 cggtcgagga gaacttctag tatatctaca tacctaatat tattgcctta ttaaaaatgg
     3781 aatcccaaca attacatcaa aatccacatt ctcttcaaaa tcaattgtcc tgtacttcct
     3841 tgttcatgtg tgttcaaaaa cgttatattt ataggataat tatactctat ttctcaacaa
     3901 gtaattggtt gtttggccga gcggtctaag gcgcctgatt caagaaatat cttgaccgca
     3961 gttaactgtg ggaatactca ggtatcgtaa gatgcaagag ttcgaatctc ttagcaacca
     4021 ttattttttt cctcaacata acgagaacac acaggggcgc tatcgcacag aatcaaattc
     4081 gatgactgga aattttttgt taatttcaga ggtcgcctga cgcatatacc tttttcaact
     4141 gaaaaattgg gagaaaaagg aaaggtgaga gcgccggaac cggcttttca tatagaatag
     4201 agaagcgttc atgactaaat gcttgcatca caatacttga agttgacaat attatttaag
     4261 gacctattgt tttttccaat aggtggttag caatcgtctt actttctaac ttttcttacc
     4321 ttttacattt cagcaatata tatatatata tttcaaggat ataccattct aatgtctgcc
     4381 cctaagaaga tcgtcgtttt gccaggtgac cacgttggtc aagaaatcac agccgaagcc
     4441 attaaggttc ttaaagctat ttctgatgtt cgttccaatg tcaagttcga tttcgaaaat
     4501 catttaattg gtggtgctgc tatcgatgct acaggtgttc cacttccaga tgaggcgctg
     4561 gaagcctcca agaaggctga tgccgttttg ttaggtgctg tgggtggtcc taaatggggt
     4621 accggtagtg ttagacctga acaaggttta ctaaaaatcc gtaaagaact tcaattgtac
     4681 gccaacttaa gaccatgtaa ctttgcatcc gactctcttt tagacttatc tccaatcaag
     4741 ccacaatttg ctaaaggtac tgacttcgtt gttgtcagag aattagtggg aggtatttac
     4801 tttggtaaga gaaaggaaga cgatggtgat ggtgtcgctt gggatagtga acaatacacc
     4861 gttccagaag tgcaaagaat cacaagaatg gccgctttca tggccctaca acatgagcca
     4921 ccattgccta tttggtcctt ggataaagct aatgttttgg cctcttcaag attatggaga
     4981 aaaactgtgg aggaaaccat caagaacgaa ttccctacat tgaaggttca acatcaattg
     5041 attgattctg ccgccatgat cctagttaag aacccaaccc acctaaatgg tattataatc
     5101 accagcaaca tgtttggtga tatcatctcc gatgaagcct ccgttatccc aggttccttg
     5161 ggtttgttgc catctgcgtc cttggcctct ttgccagaca agaacaccgc atttggtttg
     5221 tacgaaccat gccacggttc tgctccagat ttgccaaaga ataaggtcaa ccctatcgcc
     5281 actatcttgt ctgctgcaat gatgttgaaa ttgtcattga acttgcctga agaaggtaag
     5341 gccattgaag atgcagttaa aaaggttttg gatgcaggta tcagaactgg tgatttaggt
     5401 ggttccaaca gtaccaccga agtcggtgat gctgtcgccg aagaagttaa gaaaatcctt
     5461 gcttaaaaag attctctttt tttatgatat ttgtacataa actttataaa tgaaattcat
     5521 aatagaaacg acacgaaatt acaaaatgga atatgttcat agggtagacg aaactatata
     5581 cgcaatctac atacatttat caagaaggag aaaaaggagg atgtaaagga atacaggtaa
     5641 gcaaattgat actaatggct caacgtgata aggaaaaaga attgcacttt aacattaata
     5701 ttgacaagga ggagggcacc acacaaaaag ttaggtgtaa cagaaaatca tgaaactatg
     5761 attcctaatt tatatattgg aggattttct ctaaaaaaaa aaaaatacaa caaataaaaa
     5821 acactcaatg acctgaccat ttgatggagt ttaagtcaat accttcttga accatttccc
     5881 ataatggtga aagttccctc aagaatttta ctctgtcaga aacggcctta acgacgtagt
     5941 cgatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     6001 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     6061 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     6121 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     6181 taatggtttc ttaggacgga tcgcttgcct gtaacttaca cgcgcctcgt atcttttaat
     6241 gatggaataa tttgggaatt tactctgtgt ttatttattt ttatgttttg tatttggatt
     6301 ttagaaagta aataaagaag gtagaagagt tacggaatga agaaaaaaaa ataaacaaag
     6361 gtttaaaaaa tttcaacaaa aagcgtactt tacatatata tttattagac aagaaaagca
     6421 gattaaatag atatacattc gattaacgat aagtaaaatg taaaatcaca ggattttcgt
     6481 gtgtggtctt ctacacagac aagatgaaac aattcggcat taatacctga gagcaggaag
     6541 agcaagataa aaggtagtat ttgttggcga tccccctaga gtcttttaca tcttcggaaa
     6601 acaaaaacta ttttttcttt aatttctttt tttactttct atttttaatt tatatattta
     6661 tattaaaaaa tttaaattat aattattttt atagcacgtg atgaaaagga cccaggtggc
     6721 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     6781 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     6841 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     6901 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     6961 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     7021 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     7081 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     7141 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     7201 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     7261 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     7321 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     7381 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     7441 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     7501 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     7561 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     7621 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     7681 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     7741 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     7801 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     7861 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     7921 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     7981 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     8041 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     8101 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     8161 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     8221 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     8281 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     8341 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     8401 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     8461 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     8521 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     8581 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtc
//
