LOCUS       sequence-311533-        8645 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nuclear labile heme reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8645)
  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 8645)
  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: nuc-yHS1-M7A
FEATURES             Location/Qualifiers
     source          1..8645
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     31..48
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    165..186
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(201..206,207..224,225..231)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    239..255
                     /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     244..266
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     263..279
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     263..279
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     298..318
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        300..318
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        343..986
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     880..903
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             2765..2785
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2797..2813)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2798..2814)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      2816..3063
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2832..2850)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(3081..3100)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3082..3100)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3110..3127)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3110..3126)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3119..3141)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3267..3722
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3354..3373)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3564..3585
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4023..4430
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             4431..5525
                     /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(6019..6038)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6138..6160
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6198..6216)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    6258..6761
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6798..6902
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6903..6971,6972..7763)
                     /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(7121..7140)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7934..8522
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8423..8442
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
       61 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
      121 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      181 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      241 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
      301 attaaccctc actaaaggga acaaaagctg gagctcagtt tatcattatc aatactcgcc
      361 atttcaaaga atacgtaaat aattaatagt agtgattttc ctaactttat ttagtcaaaa
      421 aattagcctt ttaattctgc tgtaacccgt acatgcccaa aatagggggc gggttacaca
      481 gaatatataa catcgtaggt gtctgggtga acagtttatt cctggcatcc actaaatata
      541 atggagcccg ctttttaagc tggcatccag aaaaaaaaag aatcccagca ccaaaatatt
      601 gttttcttca ccaaccatca gttcataggt ccattctctt agcgcaacta cagagaacag
      661 gggcacaaac aggcaaaaaa cgggcacaac ctcaatggag tgatgcaacc tgcctggagt
      721 aaatgatgac acaaggcaat tgacccacgc atgtatctat ctcattttct tacaccttct
      781 attaccttct gctctctctg atttggaaaa agctgaaaaa aaaggttgaa accagttccc
      841 tgaaattatt cccctacttg actaataagt atataaagac ggtaggtatt gattgtaatt
      901 ctgtaaatct atttcttaaa cttcttaaat tctactttta tagttagtct tttttttagt
      961 tttaaaacac cagaacttag tttcgacgga ttctagaact agtcatatgc acatggtatc
     1021 ggaactgatc aaggaaaaca tgcacatgaa gctgtatatg gaaggcacgg tcaacaacca
     1081 ccactttaaa tgcacctcag aaggcgaagg taaaccttat gaaggtaccc aaactatgag
     1141 aattaaggct gtcgaaggtg gcccactgcc gtttgccttc gatatcttag cgacttcttt
     1201 tatgtacggc tcaaagacat tcatcaacca tacgcaaggt atcccagatt tctttaaaca
     1261 gtcctttccg gaaggcttca cctgggaacg cgttaccact tatgaagacg gtggcgtgct
     1321 gacagctacg caagatactt cgttacagga cggttgtttg atttacaacg tcaaaatccg
     1381 tggcgtaaat tttcctagca acggtccagt catgcagaaa aagacattag gctgggaagc
     1441 tagtaccgaa actttgtatc cggcagatgg tggcctggaa ggtcgcgctg acatggcact
     1501 gaaattagta ggtggcggtc acttgatttg caatctgaag acaacgtacc gtagtaaaaa
     1561 gccggccaaa aacttaaaga tgcctggtgt ttattacgtg gatcgtagat tggaaagaat
     1621 caaagaagcg gacaaggaaa cgtatgtgga acagcacgaa gtcgccgtcg cacgctactg
     1681 tgatttaccg tccaagttag gtcatagagg tggatccatg gtgtccaaag gtgaagaact
     1741 gtttactggt gtagtgccga ttttagttga attagacggt gatgtgaatg gtcataagtt
     1801 tagcgtgtcg ggtgaaggcg aaggtgatgc tacctttggt ggctctgcag atctggaaga
     1861 caacgcggaa actctgaacg ataacttgaa ggttatcgaa aaggccgata atgctgcaca
     1921 agtgaaagac gcgttaacaa agatgcgcgc cgcggctttg gatgctcaga aagcaacgcc
     1981 accgaaactg gaggataagt ctcctgactc accagaaatg aaagattttc gtcatggctt
     2041 cgacatttta gtgggtcaaa tcgatgacgc cttgaaactg gcgaacgagg gtaaagtcaa
     2101 ggaagctcaa gcagccgcgg aacagctgaa gaccactcgt aatgcatatc atcagaaata
     2161 cagaggtggc aaactgacct tgaagtttat ctgtacaacg ggtaaactgc ctgttccatg
     2221 gccgactttg gtgaccactc tgggctatgg tgtacaatgc ttctcacgtt acccagatca
     2281 tatgaaacag cacgactttt tcaagagcgc catgccggaa ggctacgttc aagaacgcac
     2341 tattttcttt aaggatgacg gtaactacaa gacacgtgcg gaagtcaaat ttgaaggcga
     2401 tacgctggta aacagaatcg aattgaaggg tatcgatttc aaggaagacg gcaacatctt
     2461 gggtcataaa ttggaataca actacaactc ccacaacgtc tacatcatgg ctgataagca
     2521 aaagaatggc atcaaggtaa acttcaagat cagacataac atcgaagatg gttctgttca
     2581 gctggctgac cactatcaac agaatacacc tattggcgat ggtccggtgc tgttacctga
     2641 taaccattac ttgagcacgc agagtgcact gtccaaagat ccaaatgaaa agcgcgacca
     2701 catggtcttg ttggaatttg tcaccgctgc tggtatcact ttgggtatgg atgaactgta
     2761 taaacctaag aagaagagga aggtttgaaa gcttatcgat accgtcgacc tcgagtcatg
     2821 taattagtta tgtcacgctt acattcacgc cctcccccca catccgctct aaccgaaaag
     2881 gaaggagtta gacaacctga agtctaggtc cctatttatt tttttatagt tatgttagta
     2941 ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacagacg cgtgtacgca
     3001 tgtaacatta tactgaaaac cttgcttgag aaggttttgg gacgctcgaa ggctttaatt
     3061 tgcggccggt acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg
     3121 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     3181 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     3241 agttgcgcag cctgaatggc gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg
     3301 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3361 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3421 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3481 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3541 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3601 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3661 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     3721 ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatcgacc
     3781 ggtcgaggag aacttctagt atatctacat acctaatatt attgccttat taaaaatgga
     3841 atcccaacaa ttacatcaaa atccacattc tcttcaaaat caattgtcct gtacttcctt
     3901 gttcatgtgt gttcaaaaac gttatattta taggataatt atactctatt tctcaacaag
     3961 taattggttg tttggccgag cggtctaagg cgcctgattc aagaaatatc ttgaccgcag
     4021 ttaactgtgg gaatactcag gtatcgtaag atgcaagagt tcgaatctct tagcaaccat
     4081 tatttttttc ctcaacataa cgagaacaca caggggcgct atcgcacaga atcaaattcg
     4141 atgactggaa attttttgtt aatttcagag gtcgcctgac gcatatacct ttttcaactg
     4201 aaaaattggg agaaaaagga aaggtgagag cgccggaacc ggcttttcat atagaataga
     4261 gaagcgttca tgactaaatg cttgcatcac aatacttgaa gttgacaata ttatttaagg
     4321 acctattgtt ttttccaata ggtggttagc aatcgtctta ctttctaact tttcttacct
     4381 tttacatttc agcaatatat atatatatat ttcaaggata taccattcta atgtctgccc
     4441 ctaagaagat cgtcgttttg ccaggtgacc acgttggtca agaaatcaca gccgaagcca
     4501 ttaaggttct taaagctatt tctgatgttc gttccaatgt caagttcgat ttcgaaaatc
     4561 atttaattgg tggtgctgct atcgatgcta caggtgttcc acttccagat gaggcgctgg
     4621 aagcctccaa gaaggctgat gccgttttgt taggtgctgt gggtggtcct aaatggggta
     4681 ccggtagtgt tagacctgaa caaggtttac taaaaatccg taaagaactt caattgtacg
     4741 ccaacttaag accatgtaac tttgcatccg actctctttt agacttatct ccaatcaagc
     4801 cacaatttgc taaaggtact gacttcgttg ttgtcagaga attagtggga ggtatttact
     4861 ttggtaagag aaaggaagac gatggtgatg gtgtcgcttg ggatagtgaa caatacaccg
     4921 ttccagaagt gcaaagaatc acaagaatgg ccgctttcat ggccctacaa catgagccac
     4981 cattgcctat ttggtccttg gataaagcta atgttttggc ctcttcaaga ttatggagaa
     5041 aaactgtgga ggaaaccatc aagaacgaat tccctacatt gaaggttcaa catcaattga
     5101 ttgattctgc cgccatgatc ctagttaaga acccaaccca cctaaatggt attataatca
     5161 ccagcaacat gtttggtgat atcatctccg atgaagcctc cgttatccca ggttccttgg
     5221 gtttgttgcc atctgcgtcc ttggcctctt tgccagacaa gaacaccgca tttggtttgt
     5281 acgaaccatg ccacggttct gctccagatt tgccaaagaa taaggtcaac cctatcgcca
     5341 ctatcttgtc tgctgcaatg atgttgaaat tgtcattgaa cttgcctgaa gaaggtaagg
     5401 ccattgaaga tgcagttaaa aaggttttgg atgcaggtat cagaactggt gatttaggtg
     5461 gttccaacag taccaccgaa gtcggtgatg ctgtcgccga agaagttaag aaaatccttg
     5521 cttaaaaaga ttctcttttt ttatgatatt tgtacataaa ctttataaat gaaattcata
     5581 atagaaacga cacgaaatta caaaatggaa tatgttcata gggtagacga aactatatac
     5641 gcaatctaca tacatttatc aagaaggaga aaaaggagga tgtaaaggaa tacaggtaag
     5701 caaattgata ctaatggctc aacgtgataa ggaaaaagaa ttgcacttta acattaatat
     5761 tgacaaggag gagggcacca cacaaaaagt taggtgtaac agaaaatcat gaaactatga
     5821 ttcctaattt atatattgga ggattttctc taaaaaaaaa aaaatacaac aaataaaaaa
     5881 cactcaatga cctgaccatt tgatggagtt taagtcaata ccttcttgaa ccatttccca
     5941 taatggtgaa agttccctca agaattttac tctgtcagaa acggccttaa cgacgtagtc
     6001 gatatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     6061 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     6121 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     6181 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     6241 aatggtttct taggacggat cgcttgcctg taacttacac gcgcctcgta tcttttaatg
     6301 atggaataat ttgggaattt actctgtgtt tatttatttt tatgttttgt atttggattt
     6361 tagaaagtaa ataaagaagg tagaagagtt acggaatgaa gaaaaaaaaa taaacaaagg
     6421 tttaaaaaat ttcaacaaaa agcgtacttt acatatatat ttattagaca agaaaagcag
     6481 attaaataga tatacattcg attaacgata agtaaaatgt aaaatcacag gattttcgtg
     6541 tgtggtcttc tacacagaca agatgaaaca attcggcatt aatacctgag agcaggaaga
     6601 gcaagataaa aggtagtatt tgttggcgat ccccctagag tcttttacat cttcggaaaa
     6661 caaaaactat tttttcttta atttcttttt ttactttcta tttttaattt atatatttat
     6721 attaaaaaat ttaaattata attattttta tagcacgtga tgaaaaggac ccaggtggca
     6781 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     6841 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     6901 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     6961 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     7021 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     7081 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     7141 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     7201 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     7261 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     7321 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     7381 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     7441 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     7501 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     7561 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     7621 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     7681 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     7741 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     7801 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     7861 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     7921 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     7981 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     8041 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     8101 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     8161 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     8221 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     8281 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     8341 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     8401 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     8461 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     8521 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     8581 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     8641 ctgat
//
