LOCUS       sequence-311535-        8645 bp    DNA     circular SYN 15-APR-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 Apr 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,H102A
FEATURES             Location/Qualifiers
     source          1..8645
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     37..54
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    171..192
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(207..212,213..230,231..237)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    245..261
                     /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     250..272
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     269..285
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     269..285
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     304..324
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        306..324
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        349..992
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     886..909
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             2771..2791
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2803..2819)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2804..2820)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      2822..3069
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2838..2856)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(3087..3106)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3088..3106)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3116..3133)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3116..3132)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3125..3147)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3273..3728
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3360..3379)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3570..3591
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4029..4436
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             4437..5531
                     /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(6025..6044)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6144..6166
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6204..6222)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    6264..6767
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6804..6908
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6909..6977,6978..7769)
                     /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(7127..7146)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7940..8528
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8429..8448
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
       61 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
      121 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
      181 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
      241 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
      301 cgcgcaatta accctcacta aagggaacaa aagctggagc tcagtttatc attatcaata
      361 ctcgccattt caaagaatac gtaaataatt aatagtagtg attttcctaa ctttatttag
      421 tcaaaaaatt agccttttaa ttctgctgta acccgtacat gcccaaaata gggggcgggt
      481 tacacagaat atataacatc gtaggtgtct gggtgaacag tttattcctg gcatccacta
      541 aatataatgg agcccgcttt ttaagctggc atccagaaaa aaaaagaatc ccagcaccaa
      601 aatattgttt tcttcaccaa ccatcagttc ataggtccat tctcttagcg caactacaga
      661 gaacaggggc acaaacaggc aaaaaacggg cacaacctca atggagtgat gcaacctgcc
      721 tggagtaaat gatgacacaa ggcaattgac ccacgcatgt atctatctca ttttcttaca
      781 ccttctatta ccttctgctc tctctgattt ggaaaaagct gaaaaaaaag gttgaaacca
      841 gttccctgaa attattcccc tacttgacta ataagtatat aaagacggta ggtattgatt
      901 gtaattctgt aaatctattt cttaaacttc ttaaattcta cttttatagt tagtcttttt
      961 tttagtttta aaacaccaga acttagtttc gacggattct agaactagtc atatgcacat
     1021 ggtatcggaa ctgatcaagg aaaacatgca catgaagctg tatatggaag gcacggtcaa
     1081 caaccaccac tttaaatgca cctcagaagg cgaaggtaaa ccttatgaag gtacccaaac
     1141 tatgagaatt aaggctgtcg aaggtggccc actgccgttt gccttcgata tcttagcgac
     1201 ttcttttatg tacggctcaa agacattcat caaccatacg caaggtatcc cagatttctt
     1261 taaacagtcc tttccggaag gcttcacctg ggaacgcgtt accacttatg aagacggtgg
     1321 cgtgctgaca gctacgcaag atacttcgtt acaggacggt tgtttgattt acaacgtcaa
     1381 aatccgtggc gtaaattttc ctagcaacgg tccagtcatg cagaaaaaga cattaggctg
     1441 ggaagctagt accgaaactt tgtatccggc agatggtggc ctggaaggtc gcgctgacat
     1501 ggcactgaaa ttagtaggtg gcggtcactt gatttgcaat ctgaagacaa cgtaccgtag
     1561 taaaaagccg gccaaaaact taaagatgcc tggtgtttat tacgtggatc gtagattgga
     1621 aagaatcaaa gaagcggaca aggaaacgta tgtggaacag cacgaagtcg ccgtcgcacg
     1681 ctactgtgat ttaccgtcca agttaggtca tagaggtgga tccatggtgt ccaaaggtga
     1741 agaactgttt actggtgtag tgccgatttt agttgaatta gacggtgatg tgaatggtca
     1801 taagtttagc gtgtcgggtg aaggcgaagg tgatgctacc tttggtggct ctgcagatct
     1861 ggaagacaac gcggaaactc tgaacgataa cttgaaggtt atcgaaaagg ccgataatgc
     1921 tgcacaagtg aaagacgcgt taacaaagat gcgcgccgcg gctttggatg ctcagaaagc
     1981 aacgccaccg aaactggagg ataagtctcc tgactcacca gaaatgaaag attttcgtca
     2041 tggcttcgac attttagtgg gtcaaatcga tgacgccttg aaactggcga acgagggtaa
     2101 agtcaaggaa gctcaagcag ccgcggaaca gctgaagacc actcgtaatg catatgctca
     2161 gaaatacaga ggtggcaaac tgaccttgaa gtttatctgt acaacgggta aactgcctgt
     2221 tccatggccg actttggtga ccactctggg ctatggtgta caatgcttct cacgttaccc
     2281 agatcatatg aaacagcacg actttttcaa gagcgccatg ccggaaggct acgttcaaga
     2341 acgcactatt ttctttaagg atgacggtaa ctacaagaca cgtgcggaag tcaaatttga
     2401 aggcgatacg ctggtaaaca gaatcgaatt gaagggtatc gatttcaagg aagacggcaa
     2461 catcttgggt cataaattgg aatacaacta caactcccac aacgtctaca tcatggctga
     2521 taagcaaaag aatggcatca aggtaaactt caagatcaga cataacatcg aagatggttc
     2581 tgttcagctg gctgaccact atcaacagaa tacacctatt ggcgatggtc cggtgctgtt
     2641 acctgataac cattacttga gcacgcagag tgcactgtcc aaagatccaa atgaaaagcg
     2701 cgaccacatg gtcttgttgg aatttgtcac cgctgctggt atcactttgg gtatggatga
     2761 actgtataaa cctaagaaga agaggaaggt ttgaaagctt atcgataccg tcgacctcga
     2821 gtcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc cgctctaacc
     2881 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg
     2941 ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg
     3001 tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct
     3061 ttaatttgcg gccggtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg
     3121 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     3181 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     3241 cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc
     3301 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3361 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3421 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3481 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3541 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3601 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3661 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     3721 tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     3781 tcgaccggtc gaggagaact tctagtatat ctacatacct aatattattg ccttattaaa
     3841 aatggaatcc caacaattac atcaaaatcc acattctctt caaaatcaat tgtcctgtac
     3901 ttccttgttc atgtgtgttc aaaaacgtta tatttatagg ataattatac tctatttctc
     3961 aacaagtaat tggttgtttg gccgagcggt ctaaggcgcc tgattcaaga aatatcttga
     4021 ccgcagttaa ctgtgggaat actcaggtat cgtaagatgc aagagttcga atctcttagc
     4081 aaccattatt tttttcctca acataacgag aacacacagg ggcgctatcg cacagaatca
     4141 aattcgatga ctggaaattt tttgttaatt tcagaggtcg cctgacgcat ataccttttt
     4201 caactgaaaa attgggagaa aaaggaaagg tgagagcgcc ggaaccggct tttcatatag
     4261 aatagagaag cgttcatgac taaatgcttg catcacaata cttgaagttg acaatattat
     4321 ttaaggacct attgtttttt ccaataggtg gttagcaatc gtcttacttt ctaacttttc
     4381 ttacctttta catttcagca atatatatat atatatttca aggatatacc attctaatgt
     4441 ctgcccctaa gaagatcgtc gttttgccag gtgaccacgt tggtcaagaa atcacagccg
     4501 aagccattaa ggttcttaaa gctatttctg atgttcgttc caatgtcaag ttcgatttcg
     4561 aaaatcattt aattggtggt gctgctatcg atgctacagg tgttccactt ccagatgagg
     4621 cgctggaagc ctccaagaag gctgatgccg ttttgttagg tgctgtgggt ggtcctaaat
     4681 ggggtaccgg tagtgttaga cctgaacaag gtttactaaa aatccgtaaa gaacttcaat
     4741 tgtacgccaa cttaagacca tgtaactttg catccgactc tcttttagac ttatctccaa
     4801 tcaagccaca atttgctaaa ggtactgact tcgttgttgt cagagaatta gtgggaggta
     4861 tttactttgg taagagaaag gaagacgatg gtgatggtgt cgcttgggat agtgaacaat
     4921 acaccgttcc agaagtgcaa agaatcacaa gaatggccgc tttcatggcc ctacaacatg
     4981 agccaccatt gcctatttgg tccttggata aagctaatgt tttggcctct tcaagattat
     5041 ggagaaaaac tgtggaggaa accatcaaga acgaattccc tacattgaag gttcaacatc
     5101 aattgattga ttctgccgcc atgatcctag ttaagaaccc aacccaccta aatggtatta
     5161 taatcaccag caacatgttt ggtgatatca tctccgatga agcctccgtt atcccaggtt
     5221 ccttgggttt gttgccatct gcgtccttgg cctctttgcc agacaagaac accgcatttg
     5281 gtttgtacga accatgccac ggttctgctc cagatttgcc aaagaataag gtcaacccta
     5341 tcgccactat cttgtctgct gcaatgatgt tgaaattgtc attgaacttg cctgaagaag
     5401 gtaaggccat tgaagatgca gttaaaaagg ttttggatgc aggtatcaga actggtgatt
     5461 taggtggttc caacagtacc accgaagtcg gtgatgctgt cgccgaagaa gttaagaaaa
     5521 tccttgctta aaaagattct ctttttttat gatatttgta cataaacttt ataaatgaaa
     5581 ttcataatag aaacgacacg aaattacaaa atggaatatg ttcatagggt agacgaaact
     5641 atatacgcaa tctacataca tttatcaaga aggagaaaaa ggaggatgta aaggaataca
     5701 ggtaagcaaa ttgatactaa tggctcaacg tgataaggaa aaagaattgc actttaacat
     5761 taatattgac aaggaggagg gcaccacaca aaaagttagg tgtaacagaa aatcatgaaa
     5821 ctatgattcc taatttatat attggaggat tttctctaaa aaaaaaaaaa tacaacaaat
     5881 aaaaaacact caatgacctg accatttgat ggagtttaag tcaatacctt cttgaaccat
     5941 ttcccataat ggtgaaagtt ccctcaagaa ttttactctg tcagaaacgg ccttaacgac
     6001 gtagtcgata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     6061 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     6121 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     6181 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     6241 gataataatg gtttcttagg acggatcgct tgcctgtaac ttacacgcgc ctcgtatctt
     6301 ttaatgatgg aataatttgg gaatttactc tgtgtttatt tatttttatg ttttgtattt
     6361 ggattttaga aagtaaataa agaaggtaga agagttacgg aatgaagaaa aaaaaataaa
     6421 caaaggttta aaaaatttca acaaaaagcg tactttacat atatatttat tagacaagaa
     6481 aagcagatta aatagatata cattcgatta acgataagta aaatgtaaaa tcacaggatt
     6541 ttcgtgtgtg gtcttctaca cagacaagat gaaacaattc ggcattaata cctgagagca
     6601 ggaagagcaa gataaaaggt agtatttgtt ggcgatcccc ctagagtctt ttacatcttc
     6661 ggaaaacaaa aactattttt tctttaattt ctttttttac tttctatttt taatttatat
     6721 atttatatta aaaaatttaa attataatta tttttatagc acgtgatgaa aaggacccag
     6781 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     6841 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     6901 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     6961 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     7021 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     7081 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     7141 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     7201 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     7261 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     7321 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     7381 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     7441 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     7501 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     7561 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     7621 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     7681 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     7741 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     7801 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     7861 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     7921 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     7981 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     8041 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     8101 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     8161 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     8221 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     8281 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     8341 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     8401 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     8461 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     8521 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     8581 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     8641 agtga
//
