LOCUS       sequence-454991-        6164 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control cassette FIRE1, FIRE fluorescent ratiometric sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6164)
  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 6164)
  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-MC2
FEATURES             Location/Qualifiers
     source          1..6164
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        234..1412
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          465..1403
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1360..1380
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1446..1455
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1452..2162
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1597..1615)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1865..1884
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2290..2514
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2881..2915
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(3064..3082)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3103..3119)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3103..3119)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3116..3138)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3127..3143
                     /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(3151..3157,3158..3175,3176..3181))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3196..3217
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3334..3351)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3505..4093)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3585..3604)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4264..5055,5056..5124))
                     /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     4887..4906
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5125..5229)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5255..5710)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5392..5413)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5604..5623
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5837..5859
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5851..5868
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5852..5868
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5875..5894
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5875..5893
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   5960..6022
                     /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 cacgtaagcg tgaggctccg
      241 gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg
      301 tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg
      361 tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg
      421 ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg
      481 gttcccgcgg gcctggcctc tttacgggtt atggcccttg cgtgccttga attacttcca
      541 cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg tgggagagtt
      601 cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc
      661 gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct gctttcgata
      721 agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc tggcaagata
      781 gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg gccgcgggcg
      841 gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg cgagcgcggc
      901 caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg cctggcctcg
      961 cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca ccagttgcgt
     1021 gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg aggacgcggc
     1081 gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt ccgtcctcag
     1141 ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc gattagttct
     1201 cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg atggagtttc
     1261 cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg taattctcct
     1321 tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag acagtggttc
     1381 aaagtttttt tcttccattt caggtgtcgt gactataggg ggacgctgtt tgtcgacatc
     1441 gattcgccac catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca
     1501 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
     1561 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
     1621 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct
     1681 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
     1741 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
     1801 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
     1861 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc
     1921 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
     1981 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     2041 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
     2101 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt
     2161 agaagcttag atctaaaccg cccgtcttcc tccctctctt cccggcctgg cctccccctc
     2221 cttcccccgc tggccccctc cccgcaggga taatatggtc tccggcgatc gattgtacaa
     2281 gatctgtctc tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     2341 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     2401 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     2461 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggtctagc
     2521 caacaagctc gtcatcgctt tgcagaagag cagagaggat atgctcatcg tctaaagaac
     2581 tacccatttt attatatatt agtcacgata tctataacaa gaaaatatat atataataag
     2641 ttatcacgta agtagaacat gaaataacaa tataattatc gtatgagtta aatcttaaaa
     2701 gtcacgtaaa agataatcat gcgtcatttt gactcacgcg gtcgttatag ttcaaaatca
     2761 gtgacactta ccgcattgac aagcacgcct cacgggagct ccaagcggcg actgagatgt
     2821 cctaaatgca cagcgacgga ttcgcgctat ttagaaagag agagcaatat ttcaagaatg
     2881 catgcgtcaa ttttacgcag actatctttc tagggttaaa aaagatttgc gctttactcg
     2941 acctaaactt taaacacgtc atagaatctt cgtttgacaa aaaccacatt gtggccaagc
     3001 tgtgtgacgc gacgcgcgct aaagaatggc aaaccaagtc gcgcgaggta ccagcttttg
     3061 ttccctttag tgagggttaa tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt
     3121 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3181 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3241 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3301 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3361 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3421 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3481 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3541 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3601 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3661 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3721 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3781 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3841 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3901 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3961 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4021 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4081 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4141 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4201 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4261 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4321 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4381 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4441 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4501 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4561 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4621 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4681 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4741 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4801 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4861 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4921 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4981 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5041 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5101 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5161 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5221 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
     5281 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     5341 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     5401 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     5461 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     5521 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     5581 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     5641 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     5701 cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     5761 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     5821 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attgtaatac
     5881 gactcactat agggcgaatt ggagctcggt attcacgaca gcaggctgaa taataaaaaa
     5941 attagaaact attatttaac cctagaaaga taatcatatt gtgacgtacg ttaaagataa
     6001 tcatgcgtaa aattgacgca tgtgttttat cggtctgtat atcgaggttt atttattaat
     6061 ttgaatagat attaagtttt attatattta cacttacata ctaataataa attcaacaaa
     6121 caatttattt atgtttattt atttattaaa aaaaaacaaa aact
//
