LOCUS       sequence-311532-        8645 bp    DNA     circular SYN 14-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 14, 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
FEATURES             Location/Qualifiers
     source          1..8645
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     25..42
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    159..180
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(195..200,201..218,219..225)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    233..249
                     /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     238..260
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     257..273
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     257..273
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     292..312
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        294..312
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        337..980
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     874..897
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             2759..2779
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2791..2807)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2792..2808)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      2810..3057
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2826..2844)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(3075..3094)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3076..3094)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3104..3121)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3104..3120)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3113..3135)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3261..3716
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3348..3367)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3558..3579
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4017..4424
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             4425..5519
                     /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(6013..6032)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6132..6154
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6192..6210)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    6252..6755
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6792..6896
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6897..6965,6966..7757)
                     /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(7115..7134)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7928..8516
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8417..8436
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
       61 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      121 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
      181 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      241 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
      301 cctcactaaa gggaacaaaa gctggagctc agtttatcat tatcaatact cgccatttca
      361 aagaatacgt aaataattaa tagtagtgat tttcctaact ttatttagtc aaaaaattag
      421 ccttttaatt ctgctgtaac ccgtacatgc ccaaaatagg gggcgggtta cacagaatat
      481 ataacatcgt aggtgtctgg gtgaacagtt tattcctggc atccactaaa tataatggag
      541 cccgcttttt aagctggcat ccagaaaaaa aaagaatccc agcaccaaaa tattgttttc
      601 ttcaccaacc atcagttcat aggtccattc tcttagcgca actacagaga acaggggcac
      661 aaacaggcaa aaaacgggca caacctcaat ggagtgatgc aacctgcctg gagtaaatga
      721 tgacacaagg caattgaccc acgcatgtat ctatctcatt ttcttacacc ttctattacc
      781 ttctgctctc tctgatttgg aaaaagctga aaaaaaaggt tgaaaccagt tccctgaaat
      841 tattccccta cttgactaat aagtatataa agacggtagg tattgattgt aattctgtaa
      901 atctatttct taaacttctt aaattctact tttatagtta gtcttttttt tagttttaaa
      961 acaccagaac ttagtttcga cggattctag aactagtcat atgcacatgg tatcggaact
     1021 gatcaaggaa aacatgcaca tgaagctgta tatggaaggc acggtcaaca accaccactt
     1081 taaatgcacc tcagaaggcg aaggtaaacc ttatgaaggt acccaaacta tgagaattaa
     1141 ggctgtcgaa ggtggcccac tgccgtttgc cttcgatatc ttagcgactt cttttatgta
     1201 cggctcaaag acattcatca accatacgca aggtatccca gatttcttta aacagtcctt
     1261 tccggaaggc ttcacctggg aacgcgttac cacttatgaa gacggtggcg tgctgacagc
     1321 tacgcaagat acttcgttac aggacggttg tttgatttac aacgtcaaaa tccgtggcgt
     1381 aaattttcct agcaacggtc cagtcatgca gaaaaagaca ttaggctggg aagctagtac
     1441 cgaaactttg tatccggcag atggtggcct ggaaggtcgc gctgacatgg cactgaaatt
     1501 agtaggtggc ggtcacttga tttgcaatct gaagacaacg taccgtagta aaaagccggc
     1561 caaaaactta aagatgcctg gtgtttatta cgtggatcgt agattggaaa gaatcaaaga
     1621 agcggacaag gaaacgtatg tggaacagca cgaagtcgcc gtcgcacgct actgtgattt
     1681 accgtccaag ttaggtcata gaggtggatc catggtgtcc aaaggtgaag aactgtttac
     1741 tggtgtagtg ccgattttag ttgaattaga cggtgatgtg aatggtcata agtttagcgt
     1801 gtcgggtgaa ggcgaaggtg atgctacctt tggtggctct gcagatctgg aagacaacat
     1861 ggaaactctg aacgataact tgaaggttat cgaaaaggcc gataatgctg cacaagtgaa
     1921 agacgcgtta acaaagatgc gcgccgcggc tttggatgct cagaaagcaa cgccaccgaa
     1981 actggaggat aagtctcctg actcaccaga aatgaaagat tttcgtcatg gcttcgacat
     2041 tttagtgggt caaatcgatg acgccttgaa actggcgaac gagggtaaag tcaaggaagc
     2101 tcaagcagcc gcggaacagc tgaagaccac tcgtaatgca tatcatcaga aatacagagg
     2161 tggcaaactg accttgaagt ttatctgtac aacgggtaaa ctgcctgttc catggccgac
     2221 tttggtgacc actctgggct atggtgtaca atgcttctca cgttacccag atcatatgaa
     2281 acagcacgac tttttcaaga gcgccatgcc ggaaggctac gttcaagaac gcactatttt
     2341 ctttaaggat gacggtaact acaagacacg tgcggaagtc aaatttgaag gcgatacgct
     2401 ggtaaacaga atcgaattga agggtatcga tttcaaggaa gacggcaaca tcttgggtca
     2461 taaattggaa tacaactaca actcccacaa cgtctacatc atggctgata agcaaaagaa
     2521 tggcatcaag gtaaacttca agatcagaca taacatcgaa gatggttctg ttcagctggc
     2581 tgaccactat caacagaata cacctattgg cgatggtccg gtgctgttac ctgataacca
     2641 ttacttgagc acgcagagtg cactgtccaa agatccaaat gaaaagcgcg accacatggt
     2701 cttgttggaa tttgtcaccg ctgctggtat cactttgggt atggatgaac tgtataaacc
     2761 taagaagaag aggaaggttt gaaagcttat cgataccgtc gacctcgagt catgtaatta
     2821 gttatgtcac gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga
     2881 gttagacaac ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga
     2941 acgttattta tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac
     3001 attatactga aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc
     3061 cggtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac
     3121 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     3181 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     3241 gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3301 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3361 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3421 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3481 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3541 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3601 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3661 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg
     3721 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatc gaccggtcga
     3781 ggagaacttc tagtatatct acatacctaa tattattgcc ttattaaaaa tggaatccca
     3841 acaattacat caaaatccac attctcttca aaatcaattg tcctgtactt ccttgttcat
     3901 gtgtgttcaa aaacgttata tttataggat aattatactc tatttctcaa caagtaattg
     3961 gttgtttggc cgagcggtct aaggcgcctg attcaagaaa tatcttgacc gcagttaact
     4021 gtgggaatac tcaggtatcg taagatgcaa gagttcgaat ctcttagcaa ccattatttt
     4081 tttcctcaac ataacgagaa cacacagggg cgctatcgca cagaatcaaa ttcgatgact
     4141 ggaaattttt tgttaatttc agaggtcgcc tgacgcatat acctttttca actgaaaaat
     4201 tgggagaaaa aggaaaggtg agagcgccgg aaccggcttt tcatatagaa tagagaagcg
     4261 ttcatgacta aatgcttgca tcacaatact tgaagttgac aatattattt aaggacctat
     4321 tgttttttcc aataggtggt tagcaatcgt cttactttct aacttttctt accttttaca
     4381 tttcagcaat atatatatat atatttcaag gatataccat tctaatgtct gcccctaaga
     4441 agatcgtcgt tttgccaggt gaccacgttg gtcaagaaat cacagccgaa gccattaagg
     4501 ttcttaaagc tatttctgat gttcgttcca atgtcaagtt cgatttcgaa aatcatttaa
     4561 ttggtggtgc tgctatcgat gctacaggtg ttccacttcc agatgaggcg ctggaagcct
     4621 ccaagaaggc tgatgccgtt ttgttaggtg ctgtgggtgg tcctaaatgg ggtaccggta
     4681 gtgttagacc tgaacaaggt ttactaaaaa tccgtaaaga acttcaattg tacgccaact
     4741 taagaccatg taactttgca tccgactctc ttttagactt atctccaatc aagccacaat
     4801 ttgctaaagg tactgacttc gttgttgtca gagaattagt gggaggtatt tactttggta
     4861 agagaaagga agacgatggt gatggtgtcg cttgggatag tgaacaatac accgttccag
     4921 aagtgcaaag aatcacaaga atggccgctt tcatggccct acaacatgag ccaccattgc
     4981 ctatttggtc cttggataaa gctaatgttt tggcctcttc aagattatgg agaaaaactg
     5041 tggaggaaac catcaagaac gaattcccta cattgaaggt tcaacatcaa ttgattgatt
     5101 ctgccgccat gatcctagtt aagaacccaa cccacctaaa tggtattata atcaccagca
     5161 acatgtttgg tgatatcatc tccgatgaag cctccgttat cccaggttcc ttgggtttgt
     5221 tgccatctgc gtccttggcc tctttgccag acaagaacac cgcatttggt ttgtacgaac
     5281 catgccacgg ttctgctcca gatttgccaa agaataaggt caaccctatc gccactatct
     5341 tgtctgctgc aatgatgttg aaattgtcat tgaacttgcc tgaagaaggt aaggccattg
     5401 aagatgcagt taaaaaggtt ttggatgcag gtatcagaac tggtgattta ggtggttcca
     5461 acagtaccac cgaagtcggt gatgctgtcg ccgaagaagt taagaaaatc cttgcttaaa
     5521 aagattctct ttttttatga tatttgtaca taaactttat aaatgaaatt cataatagaa
     5581 acgacacgaa attacaaaat ggaatatgtt catagggtag acgaaactat atacgcaatc
     5641 tacatacatt tatcaagaag gagaaaaagg aggatgtaaa ggaatacagg taagcaaatt
     5701 gatactaatg gctcaacgtg ataaggaaaa agaattgcac tttaacatta atattgacaa
     5761 ggaggagggc accacacaaa aagttaggtg taacagaaaa tcatgaaact atgattccta
     5821 atttatatat tggaggattt tctctaaaaa aaaaaaaata caacaaataa aaaacactca
     5881 atgacctgac catttgatgg agtttaagtc aataccttct tgaaccattt cccataatgg
     5941 tgaaagttcc ctcaagaatt ttactctgtc agaaacggcc ttaacgacgt agtcgatatg
     6001 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     6061 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     6121 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     6181 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     6241 ttcttaggac ggatcgcttg cctgtaactt acacgcgcct cgtatctttt aatgatggaa
     6301 taatttggga atttactctg tgtttattta tttttatgtt ttgtatttgg attttagaaa
     6361 gtaaataaag aaggtagaag agttacggaa tgaagaaaaa aaaataaaca aaggtttaaa
     6421 aaatttcaac aaaaagcgta ctttacatat atatttatta gacaagaaaa gcagattaaa
     6481 tagatataca ttcgattaac gataagtaaa atgtaaaatc acaggatttt cgtgtgtggt
     6541 cttctacaca gacaagatga aacaattcgg cattaatacc tgagagcagg aagagcaaga
     6601 taaaaggtag tatttgttgg cgatccccct agagtctttt acatcttcgg aaaacaaaaa
     6661 ctattttttc tttaatttct ttttttactt tctattttta atttatatat ttatattaaa
     6721 aaatttaaat tataattatt tttatagcac gtgatgaaaa ggacccaggt ggcacttttc
     6781 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     6841 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     6901 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     6961 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     7021 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     7081 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     7141 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     7201 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     7261 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     7321 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     7381 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     7441 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     7501 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     7561 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     7621 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     7681 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     7741 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     7801 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7861 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7921 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7981 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     8041 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     8101 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     8161 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     8221 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     8281 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     8341 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     8401 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     8461 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     8521 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     8581 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     8641 ccgct
//
