LOCUS       sequence-457224-        6113 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sensor cassette FIRE1, FIRE fluorescent ratiometric sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6113)
  AUTHORS   Sangokoya C
  TITLE     The FIRE biosensor illuminates iron regulatory protein activity and 
            cellular iron homeostasis.
  JOURNAL   Cell Rep Methods. 2025 Jan 11:100960. doi: 
            10.1016/j.crmeth.2024.100960.
  PUBMED    39824193
REFERENCE   2  (bases 1 to 6113)
  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"; journalName: "Cell Rep 
            Methods. 2025 Jan 11:100960. doi: 10.1016/j.crmeth.2024.100960."
COMMENT     Sequence Label: FIRES-PBCS-CLV3
FEATURES             Location/Qualifiers
     source          1..6113
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     86..108
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     100..117
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     101..117
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     124..143
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        124..142
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   209..271
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    639..694
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     promoter        701..1211
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     1130..1148
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     regulatory      1253..1262
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1259..1261,1262..1264,1265..1978)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from mClover2, with improved photostability (Bajar et al., 
                     2003)"
                     /label=mClover3
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNCIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(1304..1325)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1912..1933
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2652..2876
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3243..3277
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(3426..3444)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3465..3481)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3465..3481)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3478..3500)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3489..3505
                     /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(3513..3519,3520..3537,3538..3543))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3558..3579
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3696..3713)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3867..4455)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3947..3966)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4626..5417,5418..5486))
                     /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     5249..5268
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5487..5591)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5617..6072)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5754..5775)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5966..5985
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
       61 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
      121 ttgtaatacg actcactata gggcgaattg gagctcggta ttcacgacag caggctgaat
      181 aataaaaaaa ttagaaacta ttatttaacc ctagaaagat aatcatattg tgacgtacgt
      241 taaagataat catgcgtaaa attgacgcat gtgttttatc ggtctgtata tcgaggttta
      301 tttattaatt tgaatagata ttaagtttta ttatatttac acttacatac taataataaa
      361 ttcaacaaac aatttattta tgtttattta tttattaaaa aaaaacaaaa actcaaaatt
      421 tcttctataa agtaacaaaa cttttaaaca ttctctcttt tacaaaaata aacttatttt
      481 gtactttaaa aacagtcatg ttgtattata aaataagtaa ttagcttaac ctatacataa
      541 tagaaacaaa ttatacttat tagtcagtca gaaacaactt tggcacatat caatattatg
      601 ctctcgttaa ttaagaattc gctagcggat cccaattgaa taaaggaaat ttattttcat
      661 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgag ggggttgggg ttgcgccttt
      721 tccaaggcag ccctgggttt gcgcagggac gcggctgctc tgggcgtggt tccgggaaac
      781 gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc gtcacccgga
      841 tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc tgctccgccc ctaagtcggg
      901 aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca cgtctcacta
      961 gtaccctcgc agacggacag cgccagggag caatggcagc gcgccgaccg cgatgggctg
     1021 tggccaatag cggctgctca gcagggcgcg ccgagagcag cggccgggaa ggggcggtgc
     1081 gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg tgttccgcat
     1141 tctgcaagcc tccggagcgc acgtcggcag tcggctccct cgttgaccga atcaccgacc
     1201 tctctcccca gcagatccgt cgacaccggt agcgctgttt gtcgacatcg atgccaccat
     1261 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1321 cgacgtaaac ggccacaagt tcagcgtccg cggcgagggc gagggcgatg ccaccaacgg
     1381 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1441 cgtgaccacc ttcggctacg gcgtggcctg cttcagccgc taccccgacc acatgaagca
     1501 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatctcttt
     1561 caaggacgac ggtacctaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1621 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1681 gctggagtac aacttcaaca gccactacgt ctatatcacg gccgacaagc agaagaactg
     1741 catcaaggct aacttcaaga tccgccacaa cgttgaggac ggcagcgtgc agctcgccga
     1801 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1861 cctgagccat cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1921 gctggagttc gtgaccgccg ccgggattac acatggcatg gacgagctgt acaagtaatg
     1981 tacaagatct gtttgtgttt ttttaataga atataattat cggaagcagt gccttccata
     2041 attatgacag ttatactgtc ggtttttttt aaataaaagc agcatctgct aataaaaccc
     2101 aacagatact ggaagttttg catttatggt caacacttaa gggttttaga aaacagccgt
     2161 cagccaaatg taattgaata aagttgaagc taagatttag agatgaatta aatttaatga
     2221 ggggttgcta agaagcgagc actgaccaga taagaatgct ggttttccta aatgcagtga
     2281 attgtgacca agttataaat caatgtcact taaaggctgt ggtagtactc ctgcaaaatt
     2341 ttatagctca gtttatccaa ggtgtaactc taattcccat tttgcaaaat ttccagtacc
     2401 tttgtcacaa tcctaacaca ttatcgggag cagtgtcttc cataatgtat aaagaacaag
     2461 gtagttttta cctaccacag tgtctgtatc ggagacagtg atctccatat gttacactaa
     2521 gggtgtaagt aattatcggg aacagtgttt cccataattt tcttcatgca atgacatctt
     2581 caaagcttga agatcgttag tatctaacat gtatcccaaa gatctaaact gtacaatcga
     2641 tagatctgtc tctgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2701 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2761 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     2821 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggtcta
     2881 gccaacaagc tcgtcatcgc tttgcagaag agcagagagg atatgctcat cgtctaaaga
     2941 actacccatt ttattatata ttagtcacga tatctataac aagaaaatat atatataata
     3001 agttatcacg taagtagaac atgaaataac aatataatta tcgtatgagt taaatcttaa
     3061 aagtcacgta aaagataatc atgcgtcatt ttgactcacg cggtcgttat agttcaaaat
     3121 cagtgacact taccgcattg acaagcacgc ctcacgggag ctccaagcgg cgactgagat
     3181 gtcctaaatg cacagcgacg gattcgcgct atttagaaag agagagcaat atttcaagaa
     3241 tgcatgcgtc aattttacgc agactatctt tctagggtta aaaaagattt gcgctttact
     3301 cgacctaaac tttaaacacg tcatagaatc ttcgtttgac aaaaaccaca ttgtggccaa
     3361 gctgtgtgac gcgacgcgcg ctaaagaatg gcaaaccaag tcgcgcgagg taccagcttt
     3421 tgttcccttt agtgagggtt aatttcgagc ttggcgtaat catggtcata gctgtttcct
     3481 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     3541 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     3601 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3661 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3721 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3781 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3841 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3901 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3961 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4021 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4081 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4141 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4201 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4261 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4321 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4381 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4441 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4501 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4561 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4621 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4681 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4741 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4801 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4861 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4921 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4981 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5041 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5101 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5161 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5221 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5281 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5341 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5401 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5461 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5521 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5581 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     5641 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     5701 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     5761 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     5821 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     5881 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     5941 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     6001 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     6061 tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cga
//
