LOCUS       sequence-311544-        4993 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human codon-optimized cytosolic labile heme reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4993)
  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 4993)
  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: hHS1-M7A,H102A
FEATURES             Location/Qualifiers
     source          1..4993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        113..492
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        493..696
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     647..667
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     741..760
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        741..759
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2544..2665
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2581..2600)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2635..2654
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2714..2730)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2714..2730)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2727..2749)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2738..2754
                     /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)."
     promoter        complement(join(2762..2768,2769..2786,2787..2792))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2807..2828
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2945..2962)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3116..3704)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3196..3215)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3875..4666,4667..4735))
                     /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     4498..4517
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4736..4840)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4915..4934)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
       61 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
      121 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      181 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      241 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      301 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      361 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      421 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      481 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      541 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      601 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      661 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
      721 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc tggctagcgt
      781 ttaaacttaa gcttggtacc gagctcggat ccgccaccat gcacatggtc agcgagctga
      841 tcaaggaaaa catgcacatg aaactgtaca tggaggggac tgtgaacaat caccatttca
      901 aatgcacctc cgagggcgaa gggaagccct acgagggcac acagactatg aggatcaagg
      961 cagtggaggg aggaccactg ccattcgcct ttgacattct ggctacctca ttcatgtacg
     1021 gcagcaaaac cttcatcaat cacactcagg ggattcccga cttctttaag cagtctttcc
     1081 ctgaaggctt tacttgggag cgagtgacca catacgagga tggaggcgtc ctgaccgcca
     1141 cacaggacac aagtctgcag gatggctgtc tgatctataa cgtgaagatt cgcggggtca
     1201 actttcccag taatggacct gtgatgcaga agaaaaccct gggatgggag gcttcaactg
     1261 aaaccctgta cccagcagac ggaggactgg agggacgagc agatatggct ctgaaactgg
     1321 tgggcggggg acacctgatc tgcaacctga agactaccta tcggtccaag aaacctgcta
     1381 agaatctgaa aatgccaggc gtgtactatg tggaccggag actggagaga attaaggaag
     1441 cagataaaga gacctacgtg gagcagcacg aagtggctgt cgcacgatat tgtgacctgc
     1501 cttctaaact gggccatcgg ggcgggtcta tggtgagtaa gggcgaggaa ctgttcacag
     1561 gggtggtccc aatcctggtg gaactggacg gcgatgtcaa tgggcacaag ttcagcgtgt
     1621 ccggagaggg agaaggggac gcaacctttg gaggcagcgc cgacctggaa gataatgccg
     1681 agacactgaa cgataatctg aaagtgatcg agaaagccga caacgccgct caggtcaagg
     1741 atgctctgac taaaatgagg gcagccgctc tggatgcaca gaaagccacc ccccctaagc
     1801 tggaagacaa atcacctgat agcccagaga tgaaggactt ccgccacgga tttgatatcc
     1861 tggtcggcca gattgacgat gctctgaagc tggcaaatga aggcaaggtg aaagaggcac
     1921 aggcagccgc tgagcagctg aaaacaacta ggaacgccta cgcccagaag tatcgcgggg
     1981 gaaagctgac actgaaattc atctgcacca caggcaagct gcccgtgccc tggccaactc
     2041 tggtcactac cctgggatac ggcgtgcagt gtttttcccg ctatccagac cacatgaagc
     2101 agcatgattt ctttaaatct gccatgcccg aaggctacgt gcaggagaga accatcttct
     2161 ttaaggacga tggaaactat aaaacaaggg ctgaagtgaa gttcgaggga gacactctgg
     2221 tcaaccgcat cgaactgaag ggcattgact ttaaagagga tggaaatatt ctgggccaca
     2281 agctggaata caactataat agccataacg tgtacatcat ggccgataag cagaaaaacg
     2341 gcattaaggt caatttcaaa atccggcaca atattgagga cgggagcgtg cagctggccg
     2401 atcattacca gcagaacacc ccaatcgggg acggaccagt gctgctgccc gataatcact
     2461 atctgtccac acagtctgcc ctgagtaagg accctaacga aaaaagagat cacatggtgc
     2521 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     2581 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     2641 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     2701 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     2761 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     2821 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     2881 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     2941 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3001 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3061 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3121 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3181 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3241 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3301 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3361 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3421 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3481 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3541 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3601 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3661 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3721 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3781 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3841 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3901 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3961 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4021 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4081 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4141 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4201 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4261 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4321 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4381 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4441 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4501 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4561 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4621 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4681 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4741 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4801 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4861 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
     4921 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     4981 tgagtagtgc gcg
//
