LOCUS       sequence-486370-        4812 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  Genetically encoded fluorescent iron reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4812)
  AUTHORS   Akyol A, Cimen S, Gottschalk B, Erdogan YC, Lischnig A, Barton 
            Alston A, Digigow R, Fluhmann B, Eroglu E, Graier WF, Malli R
  TITLE     Tracking Intracellular Labile Iron with a Genetically Encoded 
            Fluorescent Reporter System Based on Protein Stability.
  JOURNAL   ACS Sens. 2025 Aug 22;10(8):5854-5861. doi: 
            10.1021/acssensors.5c01165. Epub 2025 Aug 1.
  PUBMED    40747620
REFERENCE   2  (bases 1 to 4812)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssensors.5c01165"; journalName: "ACS Sens"; date: 
            "2025-08-22- 22"; volume: "10"; issue: "8"; pages: "5854-5861"
COMMENT     Sequence Label: IronFist
FEATURES             Location/Qualifiers
     source          1..4812
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..21
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        21..531
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     450..468
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     regulatory      533..542
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1748..1801
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     protein_bind    complement(2516..2540)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2569..2690
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2580..2599)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2634..2653
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2806..2823)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2977..3565)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3057..3076)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3736..4527,4528..4596))
                     /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     4359..4378
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4597..4701)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4769..4787
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 caactttgta tagaaaagtt gggatccggg ttgcgccttt tccaaggcag ccctgggttt
       61 gcgcagggac gcggctgctc tgggcgtggt tccgggaaac gcagcggcgc cgaccctggg
      121 tctcgcacat tcttcacgtc cgttcgcagc gtcacccgga tcttcgccgc tacccttgtg
      181 ggccccccgg cgacgcttcc tgctccgccc ctaagtcggg aaggttcctt gcggttcgcg
      241 gcgtgccgga cgtgacaaac ggaagccgca cgtctcacta gtaccctcgc agacggacag
      301 cgccagggag caatggcagc gcgccgaccg cgatgggctg tggccaatag cggctgctca
      361 gcagggcgcg ccgagagcag cggccgggaa ggggcggtgc gggaggcggg gtgtggggcg
      421 gtagtgtggg ccctgttcct gcccgcgcgg tgttccgcat tctgcaagcc tccggagcgc
      481 acgtcggcag tcggctccct cgttgaccga atcaccgacc tctctcccca gggccaccat
      541 ggcgcccttt cctgaagaag tggacgtctt caccgcccca cactggcgga tgaagcagct
      601 ggtggggctc tactgcgaca agctttctaa aaccaatttt tccaacaaca acgatttccg
      661 tgctcttctg cagtctttgt atgctacttt caaggagttc aaaatgcatg agcagattga
      721 aaatgaatac attattggtt tgcttcaaca acgcagccag accatttata atgtacattc
      781 tgacaataaa ctctccgaga tgcttagcct ctttgaaaag ggactgaaga atgttaagaa
      841 tgaatatgaa cagttaaatt atgcaaaaca actgaaagag agattggagg cttttacaag
      901 agattttctt cctcacatga aagaggaaga ggaggttttt cagcccatgt taatggaata
      961 ttttacctat gaagagctta aggatattaa aaagaaagtg attgcacaac actgctctgg
     1021 cggcggcgtg agtaaggggg aggaagataa catggcctca cttccagcga ctcacgaact
     1081 tcacattttt gggtccatca atggcgtgga tttcgatatg gtagggcaag ggactggcaa
     1141 ccctaacgac ggttacgaag aactcaatct gaaatcaacg aaaggggatt tgcaattttc
     1201 cccatggata ctcgtccccc atataggcta tggattccac caatatctgc cgtatccaga
     1261 cggaatgtct ccgtttcaag ccgcaatggt agacggtagc ggttaccagg tccaccggac
     1321 gatgcaattc gaagatgggg cctccctgac agtaaattat agatatacct atgaagggag
     1381 tcacataaag ggagaagctc aggtcaaagg aacgggtttc ccggctgacg gaccggtgat
     1441 gaccaattca ctgaccgccg ccgattggtg ccgctctaaa aaaacttacc caaatgataa
     1501 gactatcatt tcaactttta aatggagtta cactacgggg aatggtaaga ggtacaggtc
     1561 aacagccagg acaacttaca catttgccaa gccaatggcg gcgaactatt tgaaaaatca
     1621 gccaatgtac gttttccgaa aaaccgaatt gaaacattct aagaccgagc ttaatttcaa
     1681 ggaatggcaa aaagcattta cggatgtgat ggggatggat gagctttaca agggtaccgg
     1741 aagcggagag ggcaggggaa gtcttctaac atgcggggac gtggaggaaa atcccggccc
     1801 catggtctcc aaaggagagg aagacaatat ggccattatc aaagagttta tgcgatttaa
     1861 agtgcacatg gagggaagcg tcaacgggca tgagtttgag attgaggggg aaggagaggg
     1921 aagaccttat gaagggaccc agactgccaa actgaaagtc accaagggcg gacccctgcc
     1981 atttgcttgg gacattctga gcccacaatt catgtatggg tccaaagcct atgttaaaca
     2041 cccggcagac atacctgact acctcaagct ctcctttcct gaggggttta aatgggagag
     2101 ggtgatgaac tttgaagatg gaggcgtcgt gacagtaact caggatagca gcctgcaaga
     2161 cggggagttt atttacaagg ttaaattgcg cggaaccaac tttcccagcg atggacccgt
     2221 gatgcagaag aaaacaatgg gctgggaagc atcatccgaa agaatgtatc ccgaagatgg
     2281 cgcgttgaag ggggaaatca aacaaaggct caagttgaaa gatggagggc actatgacgc
     2341 tgaggtcaaa accacgtaca aggctaaaaa accggtacaa ctgcccggag catataatgt
     2401 taatattaag ctggacatca ccagccacaa tgaggattac acaatagtgg aacagtatga
     2461 aagagccgag ggaagacact caactggggg catggatgaa ctgtataaat gataaaccca
     2521 gctttcttgt acaaagtggt gatggccggc cgcttcgagc agacatgata agatacattg
     2581 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     2641 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     2701 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     2761 aaaacctcta caaatgtggt agcggccgcg gcgctcttcc gcttcctcgc tcactgactc
     2821 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2881 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2941 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3001 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3061 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3121 taccggatac ctgtccgcct ttctctcttc gggaagcgtg gcgctttctc atagctcacg
     3181 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3241 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3301 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3361 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3421 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3481 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3541 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3601 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3661 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3721 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3781 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3841 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3901 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3961 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4021 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4081 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4141 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4201 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4261 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4321 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4381 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4441 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4501 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4561 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4621 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4681 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     4741 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcggcg
     4801 cgccgcggcc gc
//
