LOCUS       sequence-457072-        5830 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control cassette FIRE2, FIRE fluorescent ratiometric sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5830)
  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 5830)
  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: FIREC-PBCS-MR3
FEATURES             Location/Qualifiers
     source          1..5830
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        228..1406
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          459..1397
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1354..1374
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1438..1447
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2192..2247
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   2547..2581
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(2730..2748)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2769..2785)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2769..2785)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2782..2804)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2793..2809
                     /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(2817..2823,2824..2841,2842..2847))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2862..2883
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3000..3017)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3171..3759)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3251..3270)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3930..4721,4722..4790))
                     /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     4553..4572
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4791..4895)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4921..5376)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5058..5079)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5270..5289
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5503..5525
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5517..5534
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5518..5534
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5541..5560
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5541..5559
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   5626..5688
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 caaaatttct tctataaagt aacaaaactt ttaaacattc tctcttttac aaaaataaac
       61 ttattttgta ctttaaaaac agtcatgttg tattataaaa taagtaatta gcttaaccta
      121 tacataatag aaacaaatta tacttattag tcagtcagaa acaactttgg cacatatcaa
      181 tattatgctc tcgttaatta agaattcgct agcccctgta cacgtgaggc tccggtgccc
      241 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
      301 attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
      361 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
      421 acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc
      481 gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc
      541 tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc
      601 cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg
      661 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
      721 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
      781 taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
      841 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
      901 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
      961 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
     1021 aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg
     1081 gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg
     1141 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct
     1201 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca
     1261 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
     1321 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
     1381 tttttcttcc atttcaggtg tcgtgactat agggggacgt cgcggccgcg tttgtttgcc
     1441 gccatggtgt ctaaagggga agaattgata aaggaaaata tgaggatgaa ggtagtcatg
     1501 gaggggagtg tcaacggaca tcaatttaag tgtaccggtg aaggagaagg tcgaccttat
     1561 gagggaacac aaacaatgcg aataaaagtg attgaggggg ggcctttgcc ttttgccttc
     1621 gatatattgg ctacctcatt catgtatggt tcaagaactt ttattaagta tccagcagat
     1681 attcctgact tttttaagca atcctttcca gagggtttta cttgggaacg cgtcacacgc
     1741 tacgaggatg gcggagtcgt cactgtgact caagacacca gccttgaaga tggtgaactt
     1801 gtctataatg tgaaggtaag aggagttaat ttccccagca acggacctgt gatgcagaag
     1861 aaaacaaagg gttgggaacc aaatactgaa atgatgtatc cagctgatgg gggtttgcgg
     1921 ggctacaccg acattgcact gaaggttgat ggaggtggac acctgcattg taattttgtt
     1981 actacctaca gaagcaagaa gacagttggg aatatcaaga tgcctggggt ccacgctgtg
     2041 gatcataggc tcgagcgaat tgaggagagt gacaatgaga cctatgtagt acagagagag
     2101 gtagcagtcg caaaatacag taatctgggg ggagggatgg atgagctgta taagaaccgg
     2161 tgtttaaacg ttaaccccgg gaaaccaatt gaataaagga aatttatttt cattgcaata
     2221 gtgtgttgga attttttgtg tctctcactc gagggttaac acgcgttacg taagcatcta
     2281 taacaagaaa atatatatat aataagttat cacgtaagta gaacatgaaa taacaatata
     2341 attatcgtat gagttaaatc ttaaaagtca cgtaaaagat aatcatgcgt cattttgact
     2401 cacgcggtcg ttatagttca aaatcagtga cacttaccgc attgacaagc acgcctcacg
     2461 ggagctccaa gcggcgactg agatgtccta aatgcacagc gacggattcg cgctatttag
     2521 aaagagagag caatatttca agaatgcatg cgtcaatttt acgcagacta tctttctagg
     2581 gttaaaaaag atttgcgctt tactcgacct aaactttaaa cacgtcatag aatcttcgtt
     2641 tgacaaaaac cacattgtgg ccaagctgtg tgacgcgacg cgcgctaaag aatggcaaac
     2701 caagtcgcgc gaggtaccag cttttgttcc ctttagtgag ggttaatttc gagcttggcg
     2761 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     2821 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     2881 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     2941 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3001 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3061 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3121 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3181 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3241 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3301 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3361 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3421 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3481 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3541 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3601 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3661 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3721 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3781 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3841 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3901 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3961 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4021 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4081 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4141 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4201 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4261 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4321 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4381 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4441 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4501 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4561 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4621 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4681 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4741 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4801 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4861 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4921 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     4981 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     5041 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     5101 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     5161 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     5221 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     5281 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     5341 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
     5401 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     5461 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     5521 aaacgacggc cagtgaattg taatacgact cactataggg cgaattggag ctcggtattc
     5581 acgacagcag gctgaataat aaaaaaatta gaaactatta tttaacccta gaaagataat
     5641 catattgtga cgtacgttaa agataatcat gcgtaaaatt gacgcatgtg ttttatcggt
     5701 ctgtatatcg aggtttattt attaatttga atagatatta agttttatta tatttacact
     5761 tacatactaa taataaattc aacaaacaat ttatttatgt ttatttattt attaaaaaaa
     5821 aacaaaaact
//
