LOCUS       sequence-311541-        7457 bp    DNA     circular SYN 14-APR-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 7457)
  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 7457)
  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: hHS1-M7A
FEATURES             Location/Qualifiers
     source          1..7457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        110..489
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        490..693
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     644..664
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     738..757
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        738..756
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3340..3357)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3346..3570
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3616..4044
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3703..3722)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3913..3934
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4053..4073)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4058..4387
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4238..4373
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4300..4319
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4435..4482
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4501..4875
                     /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    5005..5126
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5042..5061)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5096..5115
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5175..5191)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5175..5191)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5188..5210)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5199..5215
                     /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(5223..5229,5230..5247,5248..5253))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5268..5289
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5406..5423)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5577..6165)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5657..5676)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6336..7127,7128..7196))
                     /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     6959..6978
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7197..7301)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7376..7395)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
       61 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
      121 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      181 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      241 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      301 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      361 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      421 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      481 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      541 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      601 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      661 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      721 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      781 aacttaagct tggtaccgag ctcggatccg ccaccatgca catggtcagc gagctgatca
      841 aggaaaacat gcacatgaaa ctgtacatgg aggggactgt gaacaatcac catttcaaat
      901 gcacctccga gggcgaaggg aagccctacg agggcacaca gactatgagg atcaaggcag
      961 tggagggagg accactgcca ttcgcctttg acattctggc tacctcattc atgtacggca
     1021 gcaaaacctt catcaatcac actcagggga ttcccgactt ctttaagcag tctttccctg
     1081 aaggctttac ttgggagcga gtgaccacat acgaggatgg aggcgtcctg accgccacac
     1141 aggacacaag tctgcaggat ggctgtctga tctataacgt gaagattcgc ggggtcaact
     1201 ttcccagtaa tggacctgtg atgcagaaga aaaccctggg atgggaggct tcaactgaaa
     1261 ccctgtaccc agcagacgga ggactggagg gacgagcaga tatggctctg aaactggtgg
     1321 gcgggggaca cctgatctgc aacctgaaga ctacctatcg gtccaagaaa cctgctaaga
     1381 atctgaaaat gccaggcgtg tactatgtgg accggagact ggagagaatt aaggaagcag
     1441 ataaagagac ctacgtggag cagcacgaag tggctgtcgc acgatattgt gacctgcctt
     1501 ctaaactggg ccatcggggc gggtctatgg tgagtaaggg cgaggaactg ttcacagggg
     1561 tggtcccaat cctggtggaa ctggacggcg atgtcaatgg gcacaagttc agcgtgtccg
     1621 gagagggaga aggggacgca acctttggag gcagcgccga cctggaagat aatgccgaga
     1681 cactgaacga taatctgaaa gtgatcgaga aagccgacaa cgccgctcag gtcaaggatg
     1741 ctctgactaa aatgagggca gccgctctgg atgcacagaa agccaccccc cctaagctgg
     1801 aagacaaatc acctgatagc ccagagatga aggacttccg ccacggattt gatatcctgg
     1861 tcggccagat tgacgatgct ctgaagctgg caaatgaagg caaggtgaaa gaggcacagg
     1921 cagccgctga gcagctgaaa acaactagga acgcctacca tcagaagtat cgcgggggaa
     1981 agctgacact gaaattcatc tgcaccacag gcaagctgcc cgtgccctgg ccaactctgg
     2041 tcactaccct gggatacggc gtgcagtgtt tttcccgcta tccagaccac atgaagcagc
     2101 atgatttctt taaatctgcc atgcccgaag gctacgtgca ggagagaacc atcttcttta
     2161 aggacgatgg aaactataaa acaagggctg aagtgaagtt cgagggagac actctggtca
     2221 accgcatcga actgaagggc attgacttta aagaggatgg aaatattctg ggccacaagc
     2281 tggaatacaa ctataatagc cataacgtgt acatcatggc cgataagcag aaaaacggca
     2341 ttaaggtcaa tttcaaaatc cggcacaata ttgaggacgg gagcgtgcag ctggccgatc
     2401 attaccagca gaacacccca atcggggacg gaccagtgct gctgcccgat aatcactatc
     2461 tgtccacaca gtctgccctg agtaaggacc ctaacgaaaa aagagatcac atggtgctgc
     2521 tggagtttgt caccgcagcc gggattacac tgggaatgga cgagctgtac aagtgactcg
     2581 aggcgatgga aggcgtgcag tttaaacagt tcaactccat tactgattac cactcgctga
     2641 tgttcgatct gggcatcatc gcgcgtcgtc tgcgctccgc atctgaccgt agcaaattct
     2701 atcgtctgat cgaagcttcg ctgtatggcg ggatctccag tgccattacc cgttctctgc
     2761 gcgactacct gttgccagaa aacagcggcg tgcgtaaagc gttccaggac atggaagcgg
     2821 cgctgcgtga aaaccgtatg acgctggaag cgattcgtgt cacccagtcg gaccgcgacc
     2881 tgtttaagca tctgatcagc gaagccacca actacgtggc ggcggactac atgcgtcacg
     2941 ccaacgagcg ccgtgtccat ctcgacaaag ccctggagtt tcgtcgcgag ctacatactt
     3001 cgcgtcagca actggcggct gagcagtaca aacacgtcga tatggcgcgt gagctggcag
     3061 agcacaacgg tgccgaaggc gatctggaag cggattatca ggcggccagt gatcacctga
     3121 acctggtgca aaccgcactg cgtcagcagg agaagatcga acgctacgaa gcggatctcg
     3181 atgagctgca gatccgtctg gaagagcaaa atgaagtggt ggcagaagcc atcgaacgcc
     3241 agcaagagaa tgaggctcgt gcggaagctg ccgaactgga agtggacgag ctgtacaagt
     3301 gactcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     3361 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     3421 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     3481 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     3541 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     3601 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3661 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3721 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3781 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3841 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3901 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3961 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     4021 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     4081 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4141 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4201 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4261 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4321 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4381 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac
     4441 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
     4501 atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc
     4561 gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt
     4621 gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac
     4681 aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag
     4741 gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag
     4801 ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc
     4861 gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4921 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4981 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     5041 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5101 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     5161 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5221 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5281 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5341 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5401 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5461 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5521 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5581 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5641 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5701 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5761 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5821 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5881 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5941 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6001 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6061 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6121 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6181 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6241 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6301 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6361 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6421 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6481 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6541 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6601 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6661 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6721 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6781 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6841 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6901 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6961 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7021 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7081 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7141 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7201 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7261 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7321 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
     7381 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     7441 ctgagtagtg cgcgagc
//
