LOCUS       sequence-457221-        6404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sensor cassette FIRE2, FIRE fluorescent ratiometric sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6404)
  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 6404)
  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-E670
FEATURES             Location/Qualifiers
     source          1..6404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     11..30
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(249..353)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(379..834)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(516..537)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     728..747
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     961..983
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     975..992
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     976..992
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     999..1018
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        999..1017
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   1084..1146
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    1514..1569
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     promoter        1576..2086
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2005..2023
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     regulatory      2115..2124
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3818..4042
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4409..4443
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(4592..4610)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4631..4647)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4631..4647)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4644..4666)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4655..4671
                     /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(4679..4685,4686..4703,4704..4709))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4724..4745
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4862..4879)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5033..5621)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5113..5132)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5792..6404,1..179,180..248))
                     /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"
ORIGIN
        1 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
       61 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
      121 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
      181 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
      241 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
      301 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
      361 ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
      421 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
      481 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
      541 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
      601 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
      661 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
      721 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
      781 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt
      841 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
      901 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
      961 cccagtcacg acgttgtaaa acgacggcca gtgaattgta atacgactca ctatagggcg
     1021 aattggagct cggtattcac gacagcaggc tgaataataa aaaaattaga aactattatt
     1081 taaccctaga aagataatca tattgtgacg tacgttaaag ataatcatgc gtaaaattga
     1141 cgcatgtgtt ttatcggtct gtatatcgag gtttatttat taatttgaat agatattaag
     1201 ttttattata tttacactta catactaata ataaattcaa caaacaattt atttatgttt
     1261 atttatttat taaaaaaaaa caaaaactca aaatttcttc tataaagtaa caaaactttt
     1321 aaacattctc tcttttacaa aaataaactt attttgtact ttaaaaacag tcatgttgta
     1381 ttataaaata agtaattagc ttaacctata cataatagaa acaaattata cttattagtc
     1441 agtcagaaac aactttggca catatcaata ttatgctctc gttaattaag aattcgctag
     1501 cggatcccaa ttgaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg
     1561 tgtctctcac tcgagggggt tggggttgcg ccttttccaa ggcagccctg ggtttgcgca
     1621 gggacgcggc tgctctgggc gtggttccgg gaaacgcagc ggcgccgacc ctgggtctcg
     1681 cacattcttc acgtccgttc gcagcgtcac ccggatcttc gccgctaccc ttgtgggccc
     1741 cccggcgacg cttcctgctc cgcccctaag tcgggaaggt tccttgcggt tcgcggcgtg
     1801 ccggacgtga caaacggaag ccgcacgtct cactagtacc ctcgcagacg gacagcgcca
     1861 gggagcaatg gcagcgcgcc gaccgcgatg ggctgtggcc aatagcggct gctcagcagg
     1921 gcgcgccgag agcagcggcc gggaaggggc ggtgcgggag gcggggtgtg gggcggtagt
     1981 gtgggccctg ttcctgcccg cgcggtgttc cgcattctgc aagcctccgg agcgcacgtc
     2041 ggcagtcggc tccctcgttg accgaatcac cgacctctct ccccagcaga tccgtcgaca
     2101 ccggttcgcg acacgccgcc atggcggaag gatccgtcgc caggcagcct gacctcttga
     2161 cctgcgaaca tgaagagatc cacctcgccg gctcgatcca gccgcatggc gcgcttctgg
     2221 tcgtcagcga acatgatcat cgcgtcatcc aggccagcgc caacgccgcg gaatttctga
     2281 atctcggaag cgtactcggc gttccgctcg ccgagatcga cggcgatctg ttgatcaaga
     2341 tcctgccgca tctcgatccc accgccgaag gcatgccggt cgcggtgcgc tgccggatcg
     2401 gcaatccctc tacggagtac tgcggtctga tgcatcggcc tccggaaggc gggctgatca
     2461 tcgaactcga acgtgccggc ccgtcgatcg atctgtcagg cacgctggcg ccggcgctgg
     2521 agcggatccg cacggcgggt tcactgcgcg cgctgtgcga tgacaccgtg ctgctgtttc
     2581 agcagtgcac cggctacgac cgggtgatgg tgtatcgttt cgatgagcaa ggccacggcc
     2641 tggtattctc cgagtgccat gtgcctgggc tcgaatccta tttcggcaac cgctatccgt
     2701 cgtcgactgt cccgcagatg gcgcggcagc tgtacgtgcg gcagcgcgtc cgcgtgctgg
     2761 tcgacgtcac ctatcagccg gtgccgctgg agccgcggct gtcgccgctg accgggcgcg
     2821 atctcgacat gtcgggctgc ttcctgcgct cgatgtcgcc gtgccatctg cagttcctga
     2881 aggacatggg cgtgcgcgcc accctggcgg tgtcgctggt ggtcggcggc aagctgtggg
     2941 gcctggttgt ctgtcaccat tatctgccgc gcttcatccg tttcgagctg cgggcgatct
     3001 gcaaacggct cgccgaaagg atcgcgacgc ggatcaccgc gcttgagagc taagtgagcg
     3061 tcgacatcga ttgtacacca aagtgttaat cttaggggag agctttctgt ccttttggca
     3121 ctgagatatt tattgtttat ttatcagtga cagagttcac tataaatggt gtttttttaa
     3181 tagaatataa ttatcggaag cagtgccttc cataattatg acagttatac tgtcggtttt
     3241 ttttaaataa aagcagcatc tgctaataaa acccaacaga tactggaagt tttgcattta
     3301 tggtcaacac ttaagggttt tagaaaacag ccgtcagcca aatgtaattg aataaagttg
     3361 aagctaagat ttagagatga attaaattta attaggggtt gctaagaagc gagcactgac
     3421 cagataagaa tgctggtttt cctaaatgca gtgaattgtg accaagttat aaatcaatgt
     3481 cacttaaagg ctgtggtagt actcctgcaa aattttatag ctcagtttat ccaaggtgta
     3541 actctaattc ccattttgca aaatttccag tacctttgtc acaatcctaa cacattatcg
     3601 ggagcagtgt cttccataat gtataaagaa caaggtagtt tttacctacc acagtgtctg
     3661 tatcggagac agtgatctcc atatgttaca ctaagggtgt aagtaattat cgggaacagt
     3721 gtttcccata attttcttca tgcaatgaca tcttcaaagc ttgaagatcg ttagtatcta
     3781 acatgtatcc caaagatctg ggcgaataga tctgtctctg tgccttctag ttgccagcca
     3841 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3901 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3961 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     4021 ggggatgcgg tgggctctat ggtctagcca acaagctcgt catcgctttg cagaagagca
     4081 gagaggatat gctcatcgtc taaagaacta cccattttat tatatattag tcacgatatc
     4141 tataacaaga aaatatatat ataataagtt atcacgtaag tagaacatga aataacaata
     4201 taattatcgt atgagttaaa tcttaaaagt cacgtaaaag ataatcatgc gtcattttga
     4261 ctcacgcggt cgttatagtt caaaatcagt gacacttacc gcattgacaa gcacgcctca
     4321 cgggagctcc aagcggcgac tgagatgtcc taaatgcaca gcgacggatt cgcgctattt
     4381 agaaagagag agcaatattt caagaatgca tgcgtcaatt ttacgcagac tatctttcta
     4441 gggttaaaaa agatttgcgc tttactcgac ctaaacttta aacacgtcat agaatcttcg
     4501 tttgacaaaa accacattgt ggccaagctg tgtgacgcga cgcgcgctaa agaatggcaa
     4561 accaagtcgc gcgaggtacc agcttttgtt ccctttagtg agggttaatt tcgagcttgg
     4621 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4681 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4741 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4801 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4861 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4921 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4981 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5041 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5101 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5161 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5221 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5281 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5341 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5401 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5461 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5521 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5581 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5641 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5701 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5761 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5821 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5881 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5941 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6001 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6061 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6121 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6181 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6241 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6301 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6361 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcg
//
