LOCUS       sequence-486388-        4827 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  Iron binding deficient IronFist.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4827)
  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 4827)
  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: IronFistMUT.
FEATURES             Location/Qualifiers
     source          1..4827
                     /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        25..525
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     444..462
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     protein_bind    526..550
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      551..560
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1766..1819
                     /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(2531..2555)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2584..2705
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2595..2614)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2649..2668
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2821..2838)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2992..3580)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3072..3091)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3751..4542,4543..4611))
                     /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     4374..4393
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4612..4716)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4784..4802
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 caactttgta tagaaaagtt ggggttgcgc cttttccaag gcagccctgg gtttgcgcag
       61 ggacgcggct gctctgggcg tggttccggg aaacgcagcg gcgccgaccc tgggtctcgc
      121 acattcttca cgtccgttcg cagcgtcacc cggatcttcg ccgctaccct tgtgggcccc
      181 ccggcgacgc ttcctgctcc gcccctaagt cgggaaggtt ccttgcggtt cgcggcgtgc
      241 cggacgtgac aaacggaagc cgcacgtctc actagtaccc tcgcagacgg acagcgccag
      301 ggagcaatgg cagcgcgccg accgcgatgg gctgtggcca atagcggctg ctcagcaggg
      361 cgcgccgaga gcagcggccg ggaaggggcg gtgcgggagg cggggtgtgg ggcggtagtg
      421 tgggccctgt tcctgcccgc gcggtgttcc gcattctgca agcctccgga gcgcacgtcg
      481 gcagtcggct ccctcgttga ccgaatcacc gacctctctc cccaggcaag tttgtacaaa
      541 aaagcaggct gccaccatgg cgccctttcc tgaagaagtg gacgtcttca ccgccccagc
      601 ctggcggatg aagcagctgg tggggctcta ctgcgacaag ctttctaaaa ccaatttttc
      661 caacaacaac gatttccgtg ctcttctgca gtctttgtat gctactttca aggagttcaa
      721 aatggccgcc cagattgcca atgaatacat tattggtttg cttcaacaac gcagccagac
      781 catttataat gtacattctg acaataaact ctccgagatg cttagcctct ttgaaaaggg
      841 actgaagaat gttaagaatg aatatgaaca gttaaattat gcaaaacaac tgaaagagag
      901 attggaggct tttacaagag attttcttcc tgccatgaaa gaggccgagg aggtttttca
      961 gcccatgtta atggaatatt ttacctatga agagcttaag gatattaaaa agaaagtgat
     1021 tgcacaacac tgctctggcg gcggcgtgag taagggggag gaagataaca tggcctcact
     1081 tccagcgact cacgaacttc acatttttgg gtccatcaat ggcgtggatt tcgatatggt
     1141 agggcaaggg actggcaacc ctaacgacgg ttacgaagaa ctcaatctga aatcaacgaa
     1201 aggggatttg caattttccc catggatact cgtcccccat ataggctatg gattccacca
     1261 atatctgccg tatccagacg gaatgtctcc gtttcaagcc gcaatggtag acggtagcgg
     1321 ttaccaggtc caccggacga tgcaattcga agatggggcc tccctgacag taaattatag
     1381 atatacctat gaagggagtc acataaaggg agaagctcag gtcaaaggaa cgggtttccc
     1441 ggctgacgga ccggtgatga ccaattcact gaccgccgcc gattggtgcc gctctaaaaa
     1501 aacttaccca aatgataaga ctatcatttc aacttttaaa tggagttaca ctacggggaa
     1561 tggtaagagg tacaggtcaa cagccaggac aacttacaca tttgccaagc caatggcggc
     1621 gaactatttg aaaaatcagc caatgtacgt tttccgaaaa accgaattga aacattctaa
     1681 gaccgagctt aatttcaagg aatggcaaaa agcatttacg gatgtgatgg ggatggatga
     1741 gctttacaag ggtaccggaa gcggagaggg caggggaagt cttctaacat gcggggacgt
     1801 ggaggaaaat cccggcccca tggtctccaa aggagaggaa gacaatatgg ccattatcaa
     1861 agagtttatg cgatttaaag tgcacatgga gggaagcgtc aacgggcatg agtttgagat
     1921 tgagggggaa ggagagggaa gaccttatga agggacccag actgccaaac tgaaagtcac
     1981 caagggcgga cccctgccat ttgcttggga cattctgagc ccacaattca tgtatgggtc
     2041 caaagcctat gttaaacacc cggcagacat acctgactac ctcaagctct cctttcctga
     2101 ggggtttaaa tgggagaggg tgatgaactt tgaagatgga ggcgtcgtga cagtaactca
     2161 ggatagcagc ctgcaagacg gggagtttat ttacaaggtt aaattgcgcg gaaccaactt
     2221 tcccagcgat ggacccgtga tgcagaagaa aacaatgggc tgggaagcat catccgaaag
     2281 aatgtatccc gaagatggcg cgttgaaggg ggaaatcaaa caaaggctca agttgaaaga
     2341 tggagggcac tatgacgctg aggtcaaaac cacgtacaag gctaaaaaac cggtacaact
     2401 gcccggagca tataatgtta atattaagct ggacatcacc agccacaatg aggattacac
     2461 aatagtggaa cagtatgaaa gagccgaggg aagacactca actgggggca tggatgaact
     2521 gtataaataa acccagcttt cttgtacaaa gtggtgatgg ccggccgctt cgagcagaca
     2581 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     2641 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     2701 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     2761 ttttttaaag caagtaaaac ctctacaaat gtggtagcgg ccgcggcgct cttccgcttc
     2821 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2881 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2941 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3001 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3061 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3121 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc tcttcgggaa gcgtggcgct
     3181 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3241 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3301 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3361 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3421 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3481 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3541 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3601 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3661 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3721 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3781 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3841 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3901 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3961 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4021 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4081 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4141 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4201 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4261 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4321 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4381 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4441 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4501 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4561 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4621 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4681 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4741 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     4801 gccctttcgt cggcgcgccg cggccgc
//
