LOCUS       sequence-357579-        7944 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human PIK3R1 3` UTR region containing 1x binding site for miR-150 
            cloned downstream of luciferase reporter gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7944)
  AUTHORS   Chirichella M, Bianchi N, Dzafo E, Foli E, Gualdrini F, Kenyon A, 
            Natoli G, Monticelli S
  TITLE     RFX transcription factors control a miR-150/PDAP1 axis that 
            restrains the proliferation of human T cells.
  JOURNAL   PLoS Biol. 2022 Feb 10;20(2):e3001538. doi: 
            10.1371/journal.pbio.3001538.
  PUBMED    35143476
REFERENCE   2  (bases 1 to 7944)
  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: "PLoS 
            Biol."; date: "2022-02-10"; volume: "20(2):e3001538. doi"; pages: " 
            10.1371/journal.pbio.3001538"
COMMENT     Sequence Label: pmiRGLO Dual-Luciferase-PIK3R1-3'UTR
FEATURES             Location/Qualifiers
     source          1..7944
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      63..72
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             69..1721
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    2483..2604
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2494..2513)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2548..2567
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2849..3206
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2849..3040
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2872..2892)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3057..3192
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3251..3260
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3257..4189
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             4241..5032
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4292..4311)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4902..4921
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5096..5144
                     /note="synthetic polyadenylation signal"
     promoter        5300..5404
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5405..5473,5474..6265)
                     /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(5623..5642)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6423..7011)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6503..6522)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7127..7410
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     promoter        join(7467..7944,1..22)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7502..7525)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     7899..7918
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
ORIGIN
        1 ttcctcatct ccgggccttt cgacctgcag cccaagcttg gcaatccggt actgttggta
       61 aagccaccat ggaagatgcc aaaaacatta agaagggccc agcgccattc tacccactcg
      121 aagacgggac cgccggcgag cagctgcaca aagccatgaa gcgctacgcc ctggtgcccg
      181 gcaccatcgc ctttaccgac gcacatatcg aggtggacat tacctacgcc gagtacttcg
      241 agatgagcgt tcggctggca gaagctatga agcgctatgg gctgaataca aaccatcgga
      301 tcgtggtgtg cagcgagaat agcttgcagt tcttcatgcc cgtgttgggt gccctgttca
      361 tcggtgtggc tgtggcccca gctaacgaca tctacaacga gcgcgagctg ctgaacagca
      421 tgggcatcag ccagcccacc gtcgtattcg tgagcaagaa agggctgcaa aagatcctca
      481 acgtgcaaaa gaagctaccg atcatacaaa agatcatcat catggatagc aagaccgact
      541 accagggctt ccaaagcatg tacaccttcg tgacttccca tttgccaccc ggcttcaacg
      601 agtacgactt cgtgcccgag agcttcgacc gggacaaaac catcgccctg atcatgaaca
      661 gtagtggcag taccggattg cccaagggcg tagccctacc gcaccgcacc gcttgtgtcc
      721 gattcagtca tgcccgcgac cccatcttcg gcaaccagat catccccgac accgctatcc
      781 tcagcgtggt gccatttcac cacggcttcg gcatgttcac cacgctgggc tacttgatct
      841 gcggctttcg ggtcgtgctc atgtaccgct tcgaggagga gctattcttg cgcagcttgc
      901 aagactataa gattcaatct gccctgctgg tgcccacact atttagcttc ttcgctaaga
      961 gcactctcat cgacaagtac gacctaagca acttgcacga gatcgccagc ggcggggcgc
     1021 cgctcagcaa ggaggtaggt gaggccgtgg ccaaacgctt ccacctacca ggcatccgcc
     1081 agggctacgg cctgacagaa acaaccagcg ccattctgat cacccccgaa ggggacgaca
     1141 agcctggcgc agtaggcaag gtggtgccct tcttcgaggc taaggtggtg gacttggaca
     1201 ccggtaagac actgggtgtg aaccagcgcg gcgagctgtg cgtccgtggc cccatgatca
     1261 tgagcggcta cgttaacaac cccgaggcta caaacgctct catcgacaag gacggctggc
     1321 tgcacagcgg cgacatcgcc tactgggacg aggacgagca cttcttcatc gtggaccggc
     1381 tgaagagcct gatcaaatac aagggctacc aggtagcccc agccgaactg gagagcatcc
     1441 tgctgcaaca ccccaacatc ttcgacgccg gggtcgccgg cctgcccgac gacgatgccg
     1501 gcgagctgcc cgccgcagtc gtcgtgctgg aacacggtaa aaccatgacc gagaaggaga
     1561 tcgtggacta tgtggccagc caggttacaa ccgccaagaa gctgcgcggt ggtgttgtgt
     1621 tcgtggacga ggtgcctaaa ggactgaccg gcaagttgga cgcccgcaag atccgcgaga
     1681 ttctcattaa ggccaagaag ggcggcaaga tcgccgtgta attctagttg tttaaacgag
     1741 ctcgctagcg cctttaacca tggtgcttgt taatgctttc tgaagcttta ccagctgaaa
     1801 gttgggactc tggagagcgg aggagagaga ggcagaagaa ccctggcctg agaaggtttg
     1861 gtccagcctg gtttagcctg gatgttgctg tgcacggtgg acccagacac atcgcaccgt
     1921 ggattatttc attttgtaac aaatgaacga tatgtagcag aaaggcacgt ccactcacaa
     1981 gggacgcttt gggagaatgt cagttcatgt atgttcagaa gaaattctgt catagaaagt
     2041 gccagaaagt gtttaacttg tcaaaaaaca aaaacccagc aacagaaaaa tggagtttgg
     2101 aaaacaggac ttaaaatgac attcagtata taaaatatgt acataatatt ggatgactaa
     2161 ctatcaaata gatggatttg tatcaatacc aaatagcttc tgttttgttt tgctgaaggc
     2221 taaattcaca gcgctatgca attcttaatt ttcattaagt tgttatttca gttttaaatg
     2281 taccttcaga ataagcttcc ccaccccagt ttttgttgct tgaaaatatt gttgtcccgg
     2341 atttttgttt ctagagtcga cctgcaggca tgcaagctga tccggctgct aacaaagccc
     2401 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
     2461 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     2521 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     2581 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     2641 cagggggaga tgtgggaggt ttttttaagc aagtaaaacc tctacaaatg tggtaaaatc
     2701 gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat
     2761 ctgtgcggta tttcacaccg catacgcgga tctgcgcagc accatggcct gaaataacct
     2821 ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt
     2881 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2941 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3001 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3061 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3121 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     3181 tttttggagg cctaggcttt tgcaaaaagc ttgattcttc tgacacaaca gtctcgaacc
     3241 aaaggctgga gccaccatgg cttccaaggt gtacgacccc gagcaacgca aacgcatgat
     3301 cactgggcct cagtggtggg ctcgctgcaa gcaaatgaac gtgctggact ccttcatcaa
     3361 ctactatgat tccgagaagc acgccgagaa cgccgtgatt tttctgcatg gtaacgctgc
     3421 ctccagctac ctgtggaggc acgtcgtgcc tcacatcgag cccgtggcta gatgcatcat
     3481 ccctgatctg atcggaatgg gtaagtccgg caagagcggg aatggctcat atcgcctcct
     3541 ggatcactac aagtacctca ccgcttggtt cgagctgctg aaccttccaa agaaaatcat
     3601 ctttgtgggc cacgactggg gggcttgtct ggcctttcac tactcctacg agcaccaaga
     3661 caagatcaag gccatcgtcc atgctgagag tgtcgtggac gtgatcgagt cctgggacga
     3721 gtggcctgac atcgaggagg atatcgccct gatcaagagc gaagagggcg agaaaatggt
     3781 gcttgagaat aacttcttcg tcgagaccat gctcccaagc aagatcatgc ggaaactgga
     3841 gcctgaggag ttcgctgcct acctggagcc attcaaggag aagggcgagg ttagacggcc
     3901 taccctctcc tggcctcgcg agatccctct cgttaaggga ggcaagcccg acgtcgtcca
     3961 gattgtccgc aactacaacg cctaccttcg ggccagcgac gatctgccta agatgttcat
     4021 cgagtccgac cctgggttct tttccaacgc tattgtcgag ggagctaaga agttccctaa
     4081 caccgagttc gtgaaggtga agggcctcca cttcagccag gaggacgctc cagatgaaat
     4141 gggtaagtac atcaagagct tcgtggagcg cgtgctgaag aacgagcaga ccggtggtgg
     4201 gagcggaggt ggcggatcag gtggcggagg ctccggaggg attgaacaag atggattgca
     4261 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4321 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4381 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4441 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4501 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4561 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4621 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4681 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4741 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4801 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4861 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4921 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4981 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5041 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgatg gccgcaataa
     5101 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagcgataag
     5161 gatcctcttt gcgcttgcgt tttcccttgt ccagatagcc cagtagctga cattcatccg
     5221 gggtcagcac cgtttctgcg gactggcttt ctacgtaatg gtttcttaga cgtcaggtgg
     5281 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5341 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5401 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5461 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5521 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5581 ccccgaagaa cgttttccaa tgatgagcac tttcaaagtt ctgctatgtg gcgcggtatt
     5641 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5701 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5761 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5821 tatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5881 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5941 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     6001 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     6061 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     6121 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     6181 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     6241 tgcctcactg attaagcatt ggtaattcga aatgaccgac caagcgacgc ccaaccggta
     6301 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     6361 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6421 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6481 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6541 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6601 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6661 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6721 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6781 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6841 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     6901 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6961 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatttca agaagatcct
     7021 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     7081 gtcatgagat tatcaaaaag gatcttcacc tagatccttt tatagtccgg aaatacagga
     7141 acgcacgctg gatggccctt cgctgggatg gtgaaaccat gaaaaatggc agcttcagtg
     7201 gattaagtgg gggtaatgtg gcctgtaccc tctggttgca taggtattca tacggttaaa
     7261 atttatcagg cgcgattgcg gcagtttttc gggtggtttg ttgccatttt tacctgtctg
     7321 ctgccgtgat cgcgctgaac gcgttttagc ggtgcgtaca attaagggat tatggtaaat
     7381 ccacttactg tctgccctcg tagccatcga gataaaccgc agtactccgg ccacgatgcg
     7441 tccggcgtag aggatcgaga tctaccgggt aggggaggcg cttttcccaa ggcagtctgg
     7501 agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg
     7561 cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc
     7621 gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg
     7681 caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc
     7741 tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc
     7801 tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag
     7861 gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg
     7921 cacgtctgcc gcgctgttct cctc
//
