LOCUS       sequence-486384-        4362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Iron Insensitive Control Construct under the same expression 
            promoter used in IronFist.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4362)
  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 4362)
  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"; doi: 
            "10.1021/acssensors.5c01165"; journalName: "ACS Sens"; date: 
            "2025-08-22- 22"; volume: "10"; issue: "8"; pages: "5854-5861"
COMMENT     Sequence Label: Iron Insensitive Control Construct 2
FEATURES             Location/Qualifiers
     source          1..4362
                     /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      557..566
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1286..1339
                     /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(2066..2090)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2119..2240
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2130..2149)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2184..2203
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2356..2373)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2527..3115)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2607..2626)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3286..4077,4078..4146))
                     /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     3909..3928
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4147..4251)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4319..4337
                     /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 gctagcgcca ccatggtgag taagggggag gaagataaca tggcctcact
      601 tccagcgact cacgaacttc acatttttgg gtccatcaat ggcgtggatt tcgatatggt
      661 agggcaaggg actggcaacc ctaacgacgg ttacgaagaa ctcaatctga aatcaacgaa
      721 aggggatttg caattttccc catggatact cgtcccccat ataggctatg gattccacca
      781 atatctgccg tatccagacg gaatgtctcc gtttcaagcc gcaatggtag acggtagcgg
      841 ttaccaggtc caccggacga tgcaattcga agatggggcc tccctgacag taaattatag
      901 atatacctat gaagggagtc acataaaggg agaagctcag gtcaaaggaa cgggtttccc
      961 ggctgacgga ccggtgatga ccaattcact gaccgccgcc gattggtgcc gctctaaaaa
     1021 aacttaccca aatgataaga ctatcatttc aacttttaaa tggagttaca ctacggggaa
     1081 tggtaagagg tacaggtcaa cagccaggac aacttacaca tttgccaagc caatggcggc
     1141 gaactatttg aaaaatcagc caatgtacgt tttccgaaaa accgaattga aacattctaa
     1201 gaccgagctt aatttcaagg aatggcaaaa agcatttacg gatgtgatgg ggatggatga
     1261 gctttacaag gaattcggaa gcggagaggg caggggaagt cttctaacat gcggggacgt
     1321 ggaggaaaat cccggccccg gatccatggt ctccaaagga gaggaagaca atatggccat
     1381 tatcaaagag tttatgcgat ttaaagtgca catggaggga agcgtcaacg ggcatgagtt
     1441 tgagattgag ggggaaggag agggaagacc ttatgaaggg acccagactg ccaaactgaa
     1501 agtcaccaag ggcggacccc tgccatttgc ttgggacatt ctgagcccac aattcatgta
     1561 tgggtccaaa gcctatgtta aacacccggc agacatacct gactacctca agctctcctt
     1621 tcctgagggg tttaaatggg agagggtgat gaactttgaa gatggaggcg tcgtgacagt
     1681 aactcaggat agcagcctgc aagacgggga gtttatttac aaggttaaat tgcgcggaac
     1741 caactttccc agcgatggac ccgtgatgca gaagaaaaca atgggctggg aagcatcatc
     1801 cgaaagaatg tatcccgaag atggcgcgtt gaagggggaa atcaaacaaa ggctcaagtt
     1861 gaaagatgga gggcactatg acgctgaggt caaaaccacg tacaaggcta aaaaaccggt
     1921 acaactgccc ggagcatata atgttaatat taagctggac atcaccagcc acaatgagga
     1981 ttacacaata gtggaacagt atgaaagagc cgagggaaga cactcaactg ggggcatgga
     2041 tgaactgtat aaataataag gtaccaccca gctttcttgt acaaagtggt gatggccggc
     2101 cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     2161 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     2221 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     2281 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt agcggccgcg
     2341 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2401 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2461 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2521 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2581 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2641 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctctcttc
     2701 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2761 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2821 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2881 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2941 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3001 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3061 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3121 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3181 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3241 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3301 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3361 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3421 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3481 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3541 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3601 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3661 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3721 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3781 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3841 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3901 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3961 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4021 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4081 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4141 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4201 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4261 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     4321 taggcgtatc acgaggccct ttcgtcggcg cgccgcggcc gc
//
