LOCUS       sequence-311538-        6731 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 6731)
  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 6731)
  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
FEATURES             Location/Qualifiers
     source          1..6731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        141..520
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        521..724
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     675..695
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     769..788
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        769..787
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2645..2662)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2651..2875
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2921..3349
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3008..3027)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3218..3239
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3358..3378)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3363..3692
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3543..3678
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3605..3624
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3740..3787
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3806..4180
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4310..4431
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4347..4366)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4401..4420
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4480..4496)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4480..4496)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4493..4515)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4504..4520
                     /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(4528..4534,4535..4552,4553..4558))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4573..4594
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4711..4728)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4882..5470)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4962..4981)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5641..6432,6433..6501))
                     /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     6264..6283
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6502..6606)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6681..6700)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
       61 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
      121 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
      181 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      241 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      301 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      361 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      421 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      481 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      541 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      601 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      661 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      721 agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca
      781 ctatagggag acccaagctg gctagcgttt aaacttaagc ttggtaccga gctcggatcc
      841 gccaccatgc acatggtcag cgagctgatc aaggaaaaca tgcacatgaa actgtacatg
      901 gaggggactg tgaacaatca ccatttcaaa tgcacctccg agggcgaagg gaagccctac
      961 gagggcacac agactatgag gatcaaggca gtggagggag gaccactgcc attcgccttt
     1021 gacattctgg ctacctcatt catgtacggc agcaaaacct tcatcaatca cactcagggg
     1081 attcccgact tctttaagca gtctttccct gaaggcttta cttgggagcg agtgaccaca
     1141 tacgaggatg gaggcgtcct gaccgccaca caggacacaa gtctgcagga tggctgtctg
     1201 atctataacg tgaagattcg cggggtcaac tttcccagta atggacctgt gatgcagaag
     1261 aaaaccctgg gatgggaggc ttcaactgaa accctgtacc cagcagacgg aggactggag
     1321 ggacgagcag atatggctct gaaactggtg ggcgggggac acctgatctg caacctgaag
     1381 actacctatc ggtccaagaa acctgctaag aatctgaaaa tgccaggcgt gtactatgtg
     1441 gaccggagac tggagagaat taaggaagca gataaagaga cctacgtgga gcagcacgaa
     1501 gtggctgtcg cacgatattg tgacctgcct tctaaactgg gccatcgggg cgggtctatg
     1561 gtgagtaagg gcgaggaact gttcacaggg gtggtcccaa tcctggtgga actggacggc
     1621 gatgtcaatg ggcacaagtt cagcgtgtcc ggagagggag aaggggacgc aacctttgga
     1681 ggcagcgccg acctggaaga taatatggag acactgaacg ataatctgaa agtgatcgag
     1741 aaagccgaca acgccgctca ggtcaaggat gctctgacta aaatgagggc agccgctctg
     1801 gatgcacaga aagccacccc ccctaagctg gaagacaaat cacctgatag cccagagatg
     1861 aaggacttcc gccacggatt tgatatcctg gtcggccaga ttgacgatgc tctgaagctg
     1921 gcaaatgaag gcaaggtgaa agaggcacag gcagccgctg agcagctgaa aacaactagg
     1981 aacgcctacc atcagaagta tcgcggggga aagctgacac tgaaattcat ctgcaccaca
     2041 ggcaagctgc ccgtgccctg gccaactctg gtcactaccc tgggatacgg cgtgcagtgt
     2101 ttttcccgct atccagacca catgaagcag catgatttct ttaaatctgc catgcccgaa
     2161 ggctacgtgc aggagagaac catcttcttt aaggacgatg gaaactataa aacaagggct
     2221 gaagtgaagt tcgagggaga cactctggtc aaccgcatcg aactgaaggg cattgacttt
     2281 aaagaggatg gaaatattct gggccacaag ctggaataca actataatag ccataacgtg
     2341 tacatcatgg ccgataagca gaaaaacggc attaaggtca atttcaaaat ccggcacaat
     2401 attgaggacg ggagcgtgca gctggccgat cattaccagc agaacacccc aatcggggac
     2461 ggaccagtgc tgctgcccga taatcactat ctgtccacac agtctgccct gagtaaggac
     2521 cctaacgaaa aaagagatca catggtgctg ctggagtttg tcaccgcagc cgggattaca
     2581 ctgggaatgg acgagctgta caagtgactc gagtctagag ggcccgttta aacccgctga
     2641 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2701 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2761 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2821 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     2881 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     2941 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3001 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3061 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3121 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3181 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3241 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3301 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3361 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3421 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3481 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3541 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3601 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3661 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     3721 tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata
     3781 atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac
     3841 cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga
     3901 cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc
     3961 ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga
     4021 cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc
     4081 ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc
     4141 cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga
     4201 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4261 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     4321 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4381 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4441 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4501 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4561 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4621 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4681 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4741 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4801 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4861 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4921 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4981 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5041 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5101 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5161 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5221 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5281 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5341 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5401 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5461 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5521 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5581 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5641 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5701 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5761 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5821 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5881 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5941 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6001 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6061 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6121 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6181 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6241 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6301 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6361 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6421 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6481 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6541 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6601 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     6661 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     6721 ctccctgctt g
//
