LOCUS       sequence-333018-        7966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express TFR1 open reading frame. Lacks IRE regulatory elements.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7966)
  AUTHORS   Terzi EM, Sviderskiy VO, Alvarez SW, Whiten GC, Possemato R
  TITLE     Iron-sulfur cluster deficiency can be sensed by IRP2 and regulates 
            iron homeostasis and sensitivity to ferroptosis independent of IRP1 
            and FBXL5.
  JOURNAL   Sci Adv. 2021 May 26;7(22). pii: 7/22/eabg4302. doi: 
            10.1126/sciadv.abg4302. Print 2021 May.
  PUBMED    34039609
REFERENCE   2  (bases 1 to 7966)
  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: "Sci Adv."; 
            date: "2021-05-26"; volume: "7(22). pii: 7/22/eabg4302. doi"; pages:
            " 10.1126/sciadv.abg4302. Print 2021 May"
COMMENT     Sequence Label: pMXS-IRES-BLAST TFR1
FEATURES             Location/Qualifiers
     source          1..7966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     24..41
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             1600..1611
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    2632..3006
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     complement(2678..2694)
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             complement(3016..3432)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(3032..3048)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(3064..3086)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    3491..3848
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             3911..4504
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        4534..4638
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4639..4707,4708..5499)
                     /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(4857..4876)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5663..6251
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6152..6171
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6405..6422
                     /label=L4440
                     /note="L4440 vector, forward primer"
     LTR             6552..7021
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(7192..7587)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    join(7588..7596,7597..7599,7600..7966,1..208)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7761..7780)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
ORIGIN
        1 cttcacgaca ttcaacagac cttgcattcc tttggcgaga ggggaaagac ccctaggaat
       61 gctcgtcaag aagacagggc caggtttccg ggccctcaca ttgccaaaag acggcaatat
      121 ggtggaaaat aacatataga caaacgcaca ccggccttat tccaagcggc ttcggccagt
      181 aacgttaggg gggggggagg gagagggggg gggggggcgg aatttacgta gcggccgctg
      241 ttctcatgga agctatgggt atcacattta aaactcattg tcaatgtccc aaacgtcacc
      301 agagagggca tttgcagctc cctgaatagt ccaagtagct agagccaact ggtttctgaa
      361 cagcgtttca ttaaaagcac cgttattttg tttacgcagt ttcaagttct ccagtaaagc
      421 tggcagcgtg tgagagccgg agccccagaa gacatgtcgg aaaggagact cttttggaga
      481 tacgtaggga gagaggaagt gatactccac tctcatgaca cgatcattga gtttcttcat
      541 gacaaatctg tctgttttct cagcattccc gaaatctgtt gttagtctgg aagtagcacg
      601 gaagaagtct ccacgagcag aatacagcca ctgtaaactc aggcccattt cctttatgtc
      661 tgctctgtat tggttcagat ccctcacaaa tgaaagcagt tggctgttgt acctctcata
      721 gtccaggttc aattcaacat catgggttag tttaatcacg aactgaccag cgacctctgc
      781 agctgctcgt gccactttgt tcaactcagg aatcctctca atcagttcct tataggtgtc
      841 catggtggta cccaaataag gataatctgt gtcctcgcaa aaacagaaag aaactgctgg
      901 gattccagaa tatgcaagga aagggaaagc agcattgtct aaagtgagtt tctcaacttt
      961 gctggcccag ttgctgtcct gatatagaaa ttgcccagta accggatgct tcacattttg
     1021 cattgttttc tcaataagcg tatacaacag tgggctggca gaaaccttga agttgctggt
     1081 accaagaacc gctttatcca gattaatata agtgaaagcc tttaaatgca gggacgaaag
     1141 gtatccctct agccattcag tggcaccaac cgatccaaag tctccagcac tccaactggc
     1201 aaagataatg cttctgctgg gctgaaaccc atcttttaag accatatctg agaacatctg
     1261 ggcaagtttc aataggagag ctgtgcctac accggatttt gcagctccag ggccccatgc
     1321 atctctctgg gccccaacta caacatagtg atctggttct acaaagcctt taataactcc
     1381 aaagatgtta agaattttta tctctttcag cacattgctc acagtgagct tcacattctt
     1441 gctttctgag gttaccatcc tacatgtaga gtctgttttc cagtcagagg gacagtctcc
     1501 ttccatattc ccaaacagct tttctgcagc agctctggag attgtctgga caggtatatt
     1561 aggcaatcct gatgaccgag atggtggaaa ctgagtgtga ttgaaggaag ggaatccagg
     1621 tgtgtaaggg tcacctgtcc ccagatgagc atgtccaaag aatgaaagtt ctgcgttaac
     1681 aatgggaaat ttagtctggt ccatgtatat caacacacca attgcattta agctttcagc
     1741 atttgcaacc ttttctgcaa aggtgatttt ccctgctctg acaatcacta tagatccatt
     1801 cacaggagtg tataaatcct caaaatcttt tttagtacca aaattagcat ggaccagttt
     1861 accagtaact gttgcagcct tactatacgc cacataaccc ccaggattct ccaccaggta
     1921 aacaagtcta ccgttcttat caactatgat caccgagttt tgagcgctgt ctttgacctg
     1981 aatcttaaca aaatgttgat cacgccagac tttgctgagt ttaaattcac gaaattgatt
     2041 ttcaacatac aacgcaagat tttcatcttt ttgagatcca gcctcacgag ggacatatga
     2101 attttcattc agcagcttga tggtgccggt gaagtctgtg ctgtccagtt tctccgacaa
     2161 ctttctcttc aggtcatccc aatataagcg acgtgctgca gggaagtcct ctcctggctc
     2221 ctccctcact ggagactcgg ttcctgccag tctctcacac tcagtttttg gttctacccc
     2281 tttacaatag cccaagtagc caatcataaa tccaatcaag aaaaagacga tcacagcaat
     2341 agtcccatag cagatacttc cactacacct ttttggtttt gtgacattgg cctttgtgtt
     2401 attgtcagca ttttcttctt catctacagc aagtttcatc tccacatgac tgttatcgcc
     2461 atctacttgc cgagccaggc tgaaccgggt atatgacaat ggttctccac caaacaagtt
     2521 agagaatgct gatctagctt gatccatcat tctgaactgc cacacgtctc gaggcctgca
     2581 ggaattcccg taccaccaca ctgggatcct taattaacta gctagatccg gcagtctaga
     2641 ggatggtcca cccccggggt cggcagcctt cacgtgggcg gcgtgtatcc aagctgcgat
     2701 gccgtctact ttgagggcgg tgggggtggt cagcaggact gtgtaaggtc ctttccagcg
     2761 aggttctagg ttcttagtct ggtgtcggcg gacccacact gtgtcgccga ctcggtaagg
     2821 gtgaggtacc accggtcggt ccagttgttc ttggtaggct gccgccagag gtctccagac
     2881 ttcgtgctgg actaagtaga gagcctgtaa gtgagcttgg agagaggggc tgttagtaac
     2941 tcttgtcatg tcagggtcag ggaagtttac aaggggcggg ggtgccccat ataagatctc
     3001 atatggccat atgggggcgc ctagagaagg agtgagggct ggataaaggg aggatcgagg
     3061 cggggtcgaa cgaggaggtt caagggggag agacggggcg gatggaggaa gaggaggcgg
     3121 aggcttaggg tgtacaaagg gcttgaccca gggagggggg tcaaaagcca aggcttccca
     3181 ggtcacgatg taggggacct ggtctgggtg tccatgcggg ccaggtgaaa agaccttgat
     3241 cttaacctgg gtgatgaggt ctcggttaaa ggtgccgtct cgcggccatc cgacgttaaa
     3301 ggttggccat tctgcagagc agaaggtaac ccaacgtctc ttcttgacat ctaccgactg
     3361 gttgtgagcg atccgctcga catctttcca gtgacctaag gtcaaactta agggagtggt
     3421 aacagtctgg ccctaatttt cagacaaata cagaaacaca gtcagacaga gacaacacag
     3481 aacgatgctg cagcagacaa gacgcgcggc gcggcttcgg tcccaaaccg aaagcaaaaa
     3541 ttcagacgga ggcgggaact gttttaggtt ctcgtctcct accagaacca catatccctc
     3601 ctattagggg ggtgcaccaa agagtccaaa acgatcggga tttttggact caggtcgggc
     3661 cacaaaaacg gcccccgaag tccctgggac gtctcccagg gttgcggccg ggtgttccga
     3721 actcgtcagt tccaccacgg gtccgccaga tacagagcta gttagctaac tagtaccgac
     3781 gcaggcgcat aaaatcagtc atagacacta gacaatcgga cagacacaga taagttgctg
     3841 gccagcttac ctcccggtgg tgggtcggtg gtccctgggc aggggtctcc cgatcccgga
     3901 cgagccccca aatgaaagac ccccgctgac gggtagtcaa tcactcagag gagaccctcc
     3961 caaggaacag cgagaccaca agtcggatgc aactgcaaga gggtttattg gatacacggg
     4021 tacccgggcg actcagtcaa tcggaggact ggcgcgccga gtgaggggtt gtgggctctt
     4081 ttattgagct cggggagcag aagcgcgcga acagaagcga gaagcgaact gattggttag
     4141 ttcaaataag gcacagggtc atttcaggtc cttggggcac cctggaaaca tctgatggtt
     4201 ctctagaaac tgctgagggc tggaccgcat ctggggacca tctgttcttg gccctgagcc
     4261 ggggcaggaa ctgcttacca cagatatcct gtttggccca tattcagctg ttccatctgt
     4321 tcttggccct gagccggggc aggaactgct taccacagat atcctgtttg gcccatattc
     4381 agctgttcca tctgttcctg accttgatct gaacttttct attctcagtt atgtattttt
     4441 ccatgccttg caaaatggcg ttacttaagc tagcttgcca aacctacagg tggggtcttt
     4501 cattatgcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4561 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4621 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4681 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4741 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4801 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4861 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4921 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4981 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5041 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5101 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5161 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5221 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5281 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5341 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5401 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5461 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5521 tatatacttt agattgattt gcggccggcc gcaaaaggat ctaggtgaag atcctttttg
     5581 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5641 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5701 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5761 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5821 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5881 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5941 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6001 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6061 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6121 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6181 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6241 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6301 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     6361 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6421 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     6481 aatgcaacta tggccattta atgtaaatac ttaagaaaaa aaaccaaatt aattttgata
     6541 catgctgcat gtgaaagacc cccgctgacg ggtagtcaat cactcagagg agaccctccc
     6601 aaggaacagc gagaccacaa gtcggatgca actgcaagag ggtttattgg atacacgggt
     6661 acccgggcga ctcagtcaat cggaggactg gcgccccgag tgaggggttg tgggctcttt
     6721 tattgagctc ggggagcaga agcgcgcgaa cagaagcgag aagcgaactg attggttagt
     6781 tcaaataagg cacagggtca tttcaggtcc ttggggcacc ctggaaacat ctgatggttc
     6841 tctagaaact gctgagggct ggaccgcatc tggggaccat ctgttcttgg ccctgagccg
     6901 gggcaggaac tgcttaccac agatatcctg tttggcccat attcagctgt tccatctgtt
     6961 cttggccctg agccggggca ggaactgctt accacagata tcctgtttgg cccatattca
     7021 gctgttccat ctgttcctga ccttgatctg aacttctcta ttctcagtta tgtatttttc
     7081 catgccttgc aaaatggcgt tacttaagct agcttgccaa acctacaggt ggggtctttc
     7141 attcccccct ttttctggag actaaataaa atcttttatt ttatcgtcga cttagccctc
     7201 ccacacataa ccagagggca gcaattcacg aatcccaact gccgtcggct gtccatcact
     7261 gtccttcact atggctttga tcccaggatg cagatcgaga agcacctgtc ggcaccgtcc
     7321 gcaggggctc aagatgcccc tgttctcatt tccgatcgcg acgatacaag tcaggttgcc
     7381 agctgccgca gcagcagcag tgcccagcac cacgagttct gcacaaggtc ccccagtaaa
     7441 atgatataca ttgacaccag tgaagatgcg gccgtcgcta gagagagctg cgctggcgac
     7501 gctgtagtct tcagagatgg ggatgctgtt gattgtagcc gttgctcttt caatgagggt
     7561 ggattcttct tgagacaaag gaaccatggt tgtggccata ttatcatcgt gtttttcaaa
     7621 ggaaaaccac gtccccgtgg ttcggggggc ctagacgttt ttttaacctc gactaaacac
     7681 atgtaaagca tgtgcaccga ggccccagat cagatcccat acaatggggt accttctggg
     7741 catccttcag ccccttgttg aatacgcttg aggagagcca tttgactctt tccacaacta
     7801 tccaactcac aacgtggcac tggggttgtg ccgcctttgc aggtgtatct tatacacgtg
     7861 gcttttggcc gcagaggcac ctgtcgccag gtggggggtt ccgctgcctg caaagggtcg
     7921 ctacagacgt tgtttgtctt caagaagctt ccagaggaac tgcttc
//
