LOCUS       sequence-449163-        8634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human IRP2 in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8634)
  AUTHORS   Wang H, Shi H, Rajan M, Canarie ER, Hong S, Simoneschi D, Pagano M, 
            Bush MF, Stoll S, Leibold EA, Zheng N
  TITLE     FBXL5 Regulates IRP2 Stability in Iron Homeostasis via an 
            Oxygen-Responsive [2Fe2S] Cluster.
  JOURNAL   Mol Cell. 2020 Apr 2;78(1):31-41.e5. doi: 
            10.1016/j.molcel.2020.02.011. Epub 2020 Mar 2.
  PUBMED    32126207
REFERENCE   2  (bases 1 to 8634)
  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: 
            "10.1016/j.molcel.2020.02.011"; journalName: "Mol Cell"; date: 
            "2020-04-2- 2"; volume: "78"; issue: "1"; pages: "31-41.e5"
COMMENT     Sequence Label: pFB-GST-IRP2
FEATURES             Location/Qualifiers
     source          1..8634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  23..188
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      372..826
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(459..478)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     668..689
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        852..956
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(957..1025,1026..1817)
                     /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     complement(1175..1194)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1988..2576
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2477..2496
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2730..2747
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2881..3105
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(3172..3705)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3875..3880,3881..3897,3898..3903))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        4281..4372
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4297..4314
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4415..5068
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     5026..5048
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             5075..5095
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     polyA_signal    8181..8315
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8218..8237)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8272..8291
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 cctgggcggc ctaggagatc cgaaccagat aagtgaaatc tagttccaaa ctattttgtc
       61 atttttaatt ttcgtattag cttacgacgc tacacccagt tcccatctat tttgtcactc
      121 ttccctaaat aatccttaaa aactccattt ccacccctcc cagttcccaa ctattttgtc
      181 cgcccacagc ggggcatttt tcttcctgtt atgtttttaa tcaaacatcc tgccaactcc
      241 atgtgacaaa ccgtcatctt cggctacttt ttctctgtca cagaatgaaa atttttctgt
      301 catctcttcg ttattaatgt ttgtaattga ctgaatatca acgcttattt gcagcctgaa
      361 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
      421 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      481 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      541 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      601 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      661 cttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
      721 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
      781 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
      841 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
      901 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
      961 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     1021 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     1081 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     1141 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     1201 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     1261 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     1321 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     1381 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     1441 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     1501 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     1561 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     1621 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt ggctctcgcg
     1681 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     1741 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     1801 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     1861 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     1921 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     1981 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     2041 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     2101 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     2161 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     2221 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     2281 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     2341 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     2401 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     2461 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     2521 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     2581 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     2641 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     2701 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     2761 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagaccagc
     2821 cgcgtaacct ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg
     2881 tgtgggcgga caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt
     2941 cttaaactag acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat
     3001 actggacttt tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc
     3061 gtattaaaga ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa
     3121 tttaccgaac aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc
     3181 ggtacttggg tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag
     3241 agccactgcg ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc
     3301 gttggcctca tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc
     3361 gccggagact gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag
     3421 aacgtaagcc gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa
     3481 tgtcttacta cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt
     3541 ggctacgtct ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg
     3601 gtcagggccg agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt
     3661 agggcgactg ccctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac
     3721 ggcgtaacgc gcttgctgct tggatgcccg aggcatagac tgtacaaaaa aacagtcata
     3781 acaagccatg aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga
     3841 ccagttgcgt gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt
     3901 caactgggtt cgtgccttca tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag
     3961 cagcgaagtc gaggcatttc tgtcctggct ggcgaacgag cgcaaggttt cgctctccac
     4021 gcatcgtcag gcattggcgg ccttgctgtt cttctacggc aaggtgctgt gcacggatct
     4081 gccctggctt caggagatcg gaagacctcg gccgtcgcgg cgcttgccgg tggtgctgac
     4141 cccggatgaa gtggttcgca tcctcggttt tctggaaggc gagcatcgtt tgttcgccca
     4201 ggactctagc tatagttcta gtggttggct acgtattaat taaggatgga agggtgtgta
     4261 ggttaccgga atattaatag atcatggaga taattaaaat gataaccatc tcgcaaataa
     4321 ataagtattt tactgttttc gtaacagttt tgtaataaaa aaacctataa atattccgga
     4381 ttattcatac cgtcccacca tcgggcgcgg atctatgtcc cctatactag gttattggaa
     4441 aattaagggc cttgtgcaac ccactcgact tcttttggaa tatcttgaag aaaaatatga
     4501 agagcatttg tatgagcgcg atgaaggtga taaatggcga aacaaaaagt ttgaattggg
     4561 tttggagttt cccaatcttc cttattatat tgatggtgat gttaaattaa cacagtctat
     4621 ggccatcata cgttatatag ctgacaagca caacatgttg ggtggttgtc caaaagagcg
     4681 tgcagagatt tcaatgcttg aaggagcggt tttggatatt agatacggtg tttcgagaat
     4741 tgcatatagt aaagactttg aaactctcaa agttgatttt cttagcaagc tacctgaaat
     4801 gctgaaaatg ttcgaagatc gtttatgtca taaaacatat ttaaatggtg atcatgtaac
     4861 ccatcctgac ttcatgttgt atgacgctct tgatgttgtt ttatacatgg acccaatgtg
     4921 cctggatgcg ttcccaaaat tagtttgttt taaaaaacgt attgaagcta tcccacaaat
     4981 tgataagtac ttgaaatcca gcaagtatat agcatggcct ttgcagggct ggcaagccac
     5041 gtttggtggt ggcgaccatc ctccaaaatc ggatgaaaac ctctacttcc aatcgatgga
     5101 cgccccaaaa gcaggatacg cctttgagta ccttattgaa acattaaatg acagttcaca
     5161 taagaagttc ttcgatgtat ctaaacttgg caccaagtat gatgttctgc cttactcaat
     5221 acgggtcttg ttggaagctg ctgtacgaaa ttgtgatggc tttttaatga agaaggaaga
     5281 tgttatgaac attttagact ggaaaaccaa acaaagcaat gttgaagtgc cctttttccc
     5341 tgcccgtgtt cttcttcaag attttactgg aataccagca atggtggatt ttgctgctat
     5401 gagggaggca gtgaaaactc ttggaggtga tcctgagaaa gtccatcctg cttgtccgac
     5461 agatcttaca gttgaccatt ctttacaaat tgacttcagt aaatgtgcaa tacagaatgc
     5521 accaaatcct ggaggtggtg acctgcagaa agcaggaaag ctctctccac ttaaagtgca
     5581 gcctaagaag cttccctgca gaggccagac tacctgccga ggatcttgtg attctggaga
     5641 actaggccga aactcaggaa cattttcttc gcagattgag aatacaccca tcctgtgtcc
     5701 ttttcatttg caaccagtgc ctgaacctga aacagtgtta aaaaatcaag aagtagaatt
     5761 cggcagaaat cgagagaggc ttcagttttt taagtggagt tcaagagttt ttaagaatgt
     5821 ggcagtgatc cctcctggaa ctggaatggc tcatcaaata aacttagaat atttgtcaag
     5881 agtggttttt gaagaaaaag acctcctctt cccagacagt gtagtcggca cagattcaca
     5941 cataacgatg gtgaatggtt tagggattct ggggtggggg gttggaggca ttgaaacaga
     6001 agcagttatg cttggtctgc cagtttctct tactttacca gaggtggttg gatgtgagtt
     6061 aactgggtca tcaaaccctt ttgttacatc catagatgtt gttcttggta ttacaaagca
     6121 cctcaggcaa gtaggagtgg ctggaaagtt tgttgagttt tttggaagtg gagtttcaca
     6181 attatctata gttgatcgaa ctacaatagc aaacatgtgt ccggaatatg gtgctatcct
     6241 cagctttttc cctgttgaca atgtgacatt aaaacattta gaacatacag gttttagcaa
     6301 agccaaactc gaatcaatgg aaacatacct taaagctgtg aaattgtttc gaaatgacca
     6361 gaattcttca ggagaacctg aatactccca ggtgatccag attaatctga attcaatagt
     6421 tccatctgtt agtggtccaa aaagacctca ggatagagtt gctgtgacag atatgaaaag
     6481 cgatttccag gcttgcttaa atgaaaaggt tggatttaaa ggcttccaaa ttgcagctga
     6541 aaaacaaaag gatattgtct ccattcatta tgaaggaagt gaatataagc tgtctcatgg
     6601 atcagtggtc attgctgcag ttatcagttg taccaataat tgcaatccat ctgtcatgct
     6661 tgctgcaggt cttttggcta aaaaggctgt tgaagctggt ctgcgtgtta aaccttatat
     6721 aagaacaagt ttatctccag gcagtgggat ggttacacat tacctcagtt caagtggagt
     6781 attaccatat ctaagtaagc ttggatttga aatcgttggc tatggatgtt caacttgtgt
     6841 gggaaataca gcacccttat cagacgcagt tttaaatgca gtaaaacagg gtgatttggt
     6901 tacctgtgga attttatctg gaaacaaaaa ttttgaaggt cgtctttgtg attgtgttcg
     6961 tgccaattat cttgcctctc cacccttagt ggtagcttat gccatagcag gcacagtgaa
     7021 tatagatttc cagacagaac ctttaggtac tgaccccacc ggcaagaaca tttacctgca
     7081 tgatatttgg cctagtcgag aagaagttca tcgagtagag gaagaacatg ttatactatc
     7141 catgtttaaa gcattaaaag ataaaataga aatggggaat aaacggtgga attccttaga
     7201 agcaccggat tcagttttgt ttccatggga cttaaagtct acttatatca gatgcccttc
     7261 attttttgat aaacttacca aagagccaat tgcactccag gctattgaaa atgcccatgt
     7321 cttattatat ttgggagact ctgtcacaac agatcatata tcacctgcag gaagtatcgc
     7381 taggaatagt gctgccgcta agtatttgac aaacagaggc cttacccctc gtgaattcaa
     7441 ctcttacgga gctcgaagag gtaatgatgc tgtaatgaca agaggcactt ttgcaaatat
     7501 caagcttttt aataagttta ttggaaaacc agctcctaaa acaattcatt ttccatcagg
     7561 acagacgcta gatgtatttg aggctgcaga gctgtaccag aaagaaggta tcccactgat
     7621 tattttagca ggaaagaaat atggttcagg aaactccaga gactgggctg ccaaaggacc
     7681 gtatttactg ggtgtgaaag ctgttttggc tgaaagttat gaaaaaatac acaaagatca
     7741 tttgattgga attggcatag ctccacttca gttccttcca ggagaaaatg cagattcctt
     7801 gggcctctcc ggtagagaaa cattttcttt aacatttcct gaagaactgt ctcctggaat
     7861 tacattgaat atacagacaa gcactggaaa agtattcagc gtgattgctt cgtttgaaga
     7921 tgatgtggaa ataacattat acaaacatgg aggattatta aactttgtgg cacgaaaatt
     7981 ctcatgaccg gaggatgagt gtggcggccg ctttcgaatc tagagcctgc agtctcgagg
     8041 catgcggtac caagcttgtc gagaagtact agaggatcat aatcagccat accacatttg
     8101 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     8161 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     8221 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     8281 ccaaactcat caatgtatct tatcatgtct ggatctgatc actgcttgag taatcccacc
     8341 actcctctcc acttaattaa cctagaggcc gcgctggcct ctgcgaagaa gattgtgttg
     8401 aagagttccg atggtgaatc tttcgaggtt gaggaggcgg tggctctcga gtcacaaacc
     8461 atagcgcata tggttgaaga cgactgcgtc gacaacggag tccctcttcc taacgtcacg
     8521 agcaagatcc tcgccaaggt gatcgagtat tgcaagaggc acgtcgaggc tgctgcctct
     8581 aaggccgagg ccgtcgaggg tgctgctacc tccgatgacg atcttaaggc ctgg
//
