LOCUS       sequence-356142-        7948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human PDAP1 3` UTR region containing 4x binding sites 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 7948)
  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 7948)
  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-PDAP1-3'UTR
FEATURES             Location/Qualifiers
     source          1..7948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      22..31
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(72..571)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(120..139)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     513..536
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     misc_feature    628..911
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     rep_origin      1027..1615
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1516..1535
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1773..2564,2565..2633))
                     /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     2396..2415
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2634..2738)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    2894..2942
                     /note="synthetic polyadenylation signal"
     CDS             complement(3006..3797)
                     /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(3117..3136)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3727..3746
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(3849..4781)
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     regulatory      4778..4787
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4832..5189)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4846..4981
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4998..5189
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5146..5166
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    5434..5555
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5471..5490)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5525..5544
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(6321..7948,1..25))
                     /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"
ORIGIN
        1 tcttaatgtt tttggcatct tccatggtgg ctttaccaac agtaccggat tgccaagctt
       61 gggctgcagg tcgaaaggcc cggagatgag gaagaggaga acagcgcggc agacgtgcgc
      121 ttttgaagcg tgcagaatgc cgggcctccg gaggaccttc gggcgcccgc cccgcccctg
      181 agcccgcccc tgagcccgcc cccggaccca ccccttccca gcctctgagc ccagaaagcg
      241 aaggagcaaa gctgctattg gccgctgccc caaaggccta cccgcttcca ttgctcagcg
      301 gtgctgtcca tctgcacgag actagtgaga cgtgctactt ccatttgtca cgtcctgcac
      361 gacgcgagct gcggggcggg ggggaacttc ctgactaggg gaggagtaga aggtggcgcg
      421 aaggggccac caaagaacgg agccggttgg cgcctaccgg tggatgtgga atgtgtgcga
      481 ggccagaggc cacttgtgta gcgccaagtg cccagcgggg ctgctaaagc gcatgctcca
      541 gactgccttg ggaaaagcgc ctcccctacc cggtagatct cgatcctcta cgccggacgc
      601 atcgtggccg gagtactgcg gtttatctcg atggctacga gggcagacag taagtggatt
      661 taccataatc ccttaattgt acgcaccgct aaaacgcgtt cagcgcgatc acggcagcag
      721 acaggtaaaa atggcaacaa accacccgaa aaactgccgc aatcgcgcct gataaatttt
      781 aaccgtatga atacctatgc aaccagaggg tacaggccac attaccccca cttaatccac
      841 tgaagctgcc atttttcatg gtttcaccat cccagcgaag ggccatccag cgtgcgttcc
      901 tgtatttccg gactataaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
      961 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     1021 atcttcttga aatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     1081 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     1141 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     1201 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     1261 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     1321 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     1381 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     1441 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     1501 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     1561 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     1621 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     1681 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     1741 cggttgggcg tcgcttggtc ggtcatttcg aattaccaat gcttaatcag tgaggcacct
     1801 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1861 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1921 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1981 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     2041 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     2101 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     2161 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatagtt
     2221 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     2281 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     2341 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     2401 accgcgccac atagcagaac tttgaaagtg ctcatcattg gaaaacgttc ttcggggcga
     2461 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     2521 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     2581 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     2641 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     2701 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     2761 cctgacgtct aagaaaccat tacgtagaaa gccagtccgc agaaacggtg ctgaccccgg
     2821 atgaatgtca gctactgggc tatctggaca agggaaaacg caagcgcaaa gaggatcctt
     2881 atcgctatcg attcacacaa aaaaccaaca cacagatgta atgaaaataa agatatttta
     2941 ttgcggccat cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga accccagagt
     3001 cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg
     3061 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata
     3121 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg
     3181 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
     3241 gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct
     3301 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc
     3361 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga
     3421 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca
     3481 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc
     3541 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat
     3601 agccgcgctg cctcgtcctg cagttcattc agggcaccgg acaggtcggt cttgacaaaa
     3661 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc
     3721 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc
     3781 aatccatctt gttcaatccc tccggagcct ccgccacctg atccgccacc tccgctccca
     3841 ccaccggtct gctcgttctt cagcacgcgc tccacgaagc tcttgatgta cttacccatt
     3901 tcatctggag cgtcctcctg gctgaagtgg aggcccttca ccttcacgaa ctcggtgtta
     3961 gggaacttct tagctccctc gacaatagcg ttggaaaaga acccagggtc ggactcgatg
     4021 aacatcttag gcagatcgtc gctggcccga aggtaggcgt tgtagttgcg gacaatctgg
     4081 acgacgtcgg gcttgcctcc cttaacgaga gggatctcgc gaggccagga gagggtaggc
     4141 cgtctaacct cgcccttctc cttgaatggc tccaggtagg cagcgaactc ctcaggctcc
     4201 agtttccgca tgatcttgct tgggagcatg gtctcgacga agaagttatt ctcaagcacc
     4261 attttctcgc cctcttcgct cttgatcagg gcgatatcct cctcgatgtc aggccactcg
     4321 tcccaggact cgatcacgtc cacgacactc tcagcatgga cgatggcctt gatcttgtct
     4381 tggtgctcgt aggagtagtg aaaggccaga caagcccccc agtcgtggcc cacaaagatg
     4441 attttctttg gaaggttcag cagctcgaac caagcggtga ggtacttgta gtgatccagg
     4501 aggcgatatg agccattccc gctcttgccg gacttaccca ttccgatcag atcagggatg
     4561 atgcatctag ccacgggctc gatgtgaggc acgacgtgcc tccacaggta gctggaggca
     4621 gcgttaccat gcagaaaaat cacggcgttc tcggcgtgct tctcggaatc atagtagttg
     4681 atgaaggagt ccagcacgtt catttgcttg cagcgagccc accactgagg cccagtgatc
     4741 atgcgtttgc gttgctcggg gtcgtacacc ttggaagcca tggtggctcc agcctttggt
     4801 tcgagactgt tgtgtcagaa gaatcaagct ttttgcaaaa gcctaggcct ccaaaaaagc
     4861 ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa
     4921 aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga
     4981 tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac
     5041 ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca
     5101 tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca
     5161 cattccacag ctggttcttt ccgcctcaga aggtacctaa ccaagttcct ctttcagagg
     5221 ttatttcagg ccatggtgct gcgcagatcc gcgtatgcgg tgtgaaatac cgcacagatg
     5281 cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt aaaattcgat
     5341 tttaccacat ttgtagaggt tttacttgct taaaaaaacc tcccacatct ccccctgaac
     5401 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     5461 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     5521 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgctcgaa gcggccgccc
     5581 caaggggtta tgctagttat tgctcagcgg tggcagcagc caactcagct tcctttcggg
     5641 ctttgttagc agccggatca gcttgcatgc ctgcaggtcg actctagaat gcttgcctat
     5701 gtggaaggag aggttttttg ttatttcctt ggggtcttgc taagttttcc aggctgcact
     5761 agaactcgtg ggagcaatcc ttctgcctca gcctcccaag tagctgggac tccagggaga
     5821 gattggtgtt tgcacggcca taggtagata gcttattttt actgcttgcc aacaaaattg
     5881 atcctgaagc ccaacggcct tatgccaaat ggttccctcc ccactgctgt ccatcctggg
     5941 ctgccactcc caaaactgcc agtaccaggt gtcactgatt gactagtcaa attcctggac
     6001 aaaggggcta cacactccag gctggttgtg gaacttttgt tttgagatgg agttttgctc
     6061 gtcacccagg ctggagtgca gttgtgcgtt ctcagctcac tgccacctct gccttcctgg
     6121 gatcaagcga ttctcctgcc tcagcctccc aagtagctgg gattacaggc atacaccacc
     6181 aagcctggtt aatttttgta tttttagtag agacggagtt tctccatgtt ggccaggctg
     6241 gtcttgaact cctgacctca gatgatccac ccacctcacc tcccaaagtg ctgctagcga
     6301 gctcgtttaa acaactagaa ttacacggcg atcttgccgc ccttcttggc cttaatgaga
     6361 atctcgcgga tcttgcgggc gtccaacttg ccggtcagtc ctttaggcac ctcgtccacg
     6421 aacacaacac caccgcgcag cttcttggcg gttgtaacct ggctggccac atagtccacg
     6481 atctccttct cggtcatggt tttaccgtgt tccagcacga cgactgcggc gggcagctcg
     6541 ccggcatcgt cgtcgggcag gccggcgacc ccggcgtcga agatgttggg gtgttgcagc
     6601 aggatgctct ccagttcggc tggggctacc tggtagccct tgtatttgat caggctcttc
     6661 agccggtcca cgatgaagaa gtgctcgtcc tcgtcccagt aggcgatgtc gccgctgtgc
     6721 agccagccgt ccttgtcgat gagagcgttt gtagcctcgg ggttgttaac gtagccgctc
     6781 atgatcatgg ggccacggac gcacagctcg ccgcgctggt tcacacccag tgtcttaccg
     6841 gtgtccaagt ccaccacctt agcctcgaag aagggcacca ccttgcctac tgcgccaggc
     6901 ttgtcgtccc cttcgggggt gatcagaatg gcgctggttg tttctgtcag gccgtagccc
     6961 tggcggatgc ctggtaggtg gaagcgtttg gccacggcct cacctacctc cttgctgagc
     7021 ggcgccccgc cgctggcgat ctcgtgcaag ttgcttaggt cgtacttgtc gatgagagtg
     7081 ctcttagcga agaagctaaa tagtgtgggc accagcaggg cagattgaat cttatagtct
     7141 tgcaagctgc gcaagaatag ctcctcctcg aagcggtaca tgagcacgac ccgaaagccg
     7201 cagatcaagt agcccagcgt ggtgaacatg ccgaagccgt ggtgaaatgg caccacgctg
     7261 aggatagcgg tgtcggggat gatctggttg ccgaagatgg ggtcgcgggc atgactgaat
     7321 cggacacaag cggtgcggtg cggtagggct acgcccttgg gcaatccggt actgccacta
     7381 ctgttcatga tcagggcgat ggttttgtcc cggtcgaagc tctcgggcac gaagtcgtac
     7441 tcgttgaagc cgggtggcaa atgggaagtc acgaaggtgt acatgctttg gaagccctgg
     7501 tagtcggtct tgctatccat gatgatgatc ttttgtatga tcggtagctt cttttgcacg
     7561 ttgaggatct tttgcagccc tttcttgctc acgaatacga cggtgggctg gctgatgccc
     7621 atgctgttca gcagctcgcg ctcgttgtag atgtcgttag ctggggccac agccacaccg
     7681 atgaacaggg cacccaacac gggcatgaag aactgcaagc tattctcgct gcacaccacg
     7741 atccgatggt ttgtattcag cccatagcgc ttcatagctt ctgccagccg aacgctcatc
     7801 tcgaagtact cggcgtaggt aatgtccacc tcgatatgtg cgtcggtaaa ggcgatggtg
     7861 ccgggcacca gggcgtagcg cttcatggct ttgtgcagct gctcgccggc ggtcccgtct
     7921 tcgagtgggt agaatggcgc tgggccct
//
