LOCUS       sequence-359484-        7948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human PDAP1 3` UTR region mutated in 2x 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 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-Mut2x
FEATURES             Location/Qualifiers
     source          1..7948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(50..549)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(98..117)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     491..514
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     misc_feature    606..889
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     rep_origin      1005..1593
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1494..1513
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1751..2542,2543..2611))
                     /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     2374..2393
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2612..2716)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    2872..2920
                     /note="synthetic polyadenylation signal"
     CDS             complement(2984..3775)
                     /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(3095..3114)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3705..3724
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(3827..4759)
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     regulatory      4756..4765
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4810..5167)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4824..4959
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4976..5167
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5124..5144
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    5412..5533
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5449..5468)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5503..5522
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(6299..7948,1..3))
                     /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"
     regulatory      join(7948,1..9)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 catggtggct ttaccaacag taccggattg ccaagcttgg gctgcaggtc gaaaggcccg
       61 gagatgagga agaggagaac agcgcggcag acgtgcgctt ttgaagcgtg cagaatgccg
      121 ggcctccgga ggaccttcgg gcgcccgccc cgcccctgag cccgcccctg agcccgcccc
      181 cggacccacc ccttcccagc ctctgagccc agaaagcgaa ggagcaaagc tgctattggc
      241 cgctgcccca aaggcctacc cgcttccatt gctcagcggt gctgtccatc tgcacgagac
      301 tagtgagacg tgctacttcc atttgtcacg tcctgcacga cgcgagctgc ggggcggggg
      361 ggaacttcct gactagggga ggagtagaag gtggcgcgaa ggggccacca aagaacggag
      421 ccggttggcg cctaccggtg gatgtggaat gtgtgcgagg ccagaggcca cttgtgtagc
      481 gccaagtgcc cagcggggct gctaaagcgc atgctccaga ctgccttggg aaaagcgcct
      541 cccctacccg gtagatctcg atcctctacg ccggacgcat cgtggccgga gtactgcggt
      601 ttatctcgat ggctacgagg gcagacagta agtggattta ccataatccc ttaattgtac
      661 gcaccgctaa aacgcgttca gcgcgatcac ggcagcagac aggtaaaaat ggcaacaaac
      721 cacccgaaaa actgccgcaa tcgcgcctga taaattttaa ccgtatgaat acctatgcaa
      781 ccagagggta caggccacat tacccccact taatccactg aagctgccat ttttcatggt
      841 ttcaccatcc cagcgaaggg ccatccagcg tgcgttcctg tatttccgga ctataaaagg
      901 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
      961 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaaa tccttttttt
     1021 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     1081 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     1141 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     1201 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     1261 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     1321 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     1381 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     1441 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     1501 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     1561 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     1621 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     1681 gtggataacc gtattaccgc ctttgagtga gctgataccg gttgggcgtc gcttggtcgg
     1741 tcatttcgaa ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     1801 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     1861 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     1921 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     1981 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     2041 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     2101 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     2161 gcaaaaaagc ggttagctcc ttcggtcctc cgatagttgt cagaagtaag ttggccgcag
     2221 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     2281 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     2341 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     2401 tgaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     2461 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     2521 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     2581 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     2641 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     2701 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     2761 cgtagaaagc cagtccgcag aaacggtgct gaccccggat gaatgtcagc tactgggcta
     2821 tctggacaag ggaaaacgca agcgcaaaga ggatccttat cgctatcgat tcacacaaaa
     2881 aaccaacaca cagatgtaat gaaaataaag atattttatt gcggccatcg tgatggcagg
     2941 ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa
     3001 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
     3061 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt
     3121 cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat
     3181 tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt
     3241 cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt
     3301 cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc
     3361 gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca
     3421 ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct
     3481 gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca
     3541 cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca
     3601 gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg
     3661 acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga
     3721 atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatccctc
     3781 cggagcctcc gccacctgat ccgccacctc cgctcccacc accggtctgc tcgttcttca
     3841 gcacgcgctc cacgaagctc ttgatgtact tacccatttc atctggagcg tcctcctggc
     3901 tgaagtggag gcccttcacc ttcacgaact cggtgttagg gaacttctta gctccctcga
     3961 caatagcgtt ggaaaagaac ccagggtcgg actcgatgaa catcttaggc agatcgtcgc
     4021 tggcccgaag gtaggcgttg tagttgcgga caatctggac gacgtcgggc ttgcctccct
     4081 taacgagagg gatctcgcga ggccaggaga gggtaggccg tctaacctcg cccttctcct
     4141 tgaatggctc caggtaggca gcgaactcct caggctccag tttccgcatg atcttgcttg
     4201 ggagcatggt ctcgacgaag aagttattct caagcaccat tttctcgccc tcttcgctct
     4261 tgatcagggc gatatcctcc tcgatgtcag gccactcgtc ccaggactcg atcacgtcca
     4321 cgacactctc agcatggacg atggccttga tcttgtcttg gtgctcgtag gagtagtgaa
     4381 aggccagaca agccccccag tcgtggccca caaagatgat tttctttgga aggttcagca
     4441 gctcgaacca agcggtgagg tacttgtagt gatccaggag gcgatatgag ccattcccgc
     4501 tcttgccgga cttacccatt ccgatcagat cagggatgat gcatctagcc acgggctcga
     4561 tgtgaggcac gacgtgcctc cacaggtagc tggaggcagc gttaccatgc agaaaaatca
     4621 cggcgttctc ggcgtgcttc tcggaatcat agtagttgat gaaggagtcc agcacgttca
     4681 tttgcttgca gcgagcccac cactgaggcc cagtgatcat gcgtttgcgt tgctcggggt
     4741 cgtacacctt ggaagccatg gtggctccag cctttggttc gagactgttg tgtcagaaga
     4801 atcaagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
     4861 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
     4921 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac
     4981 tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     5041 ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     5101 tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagct ggttctttcc
     5161 gcctcagaag gtacctaacc aagttcctct ttcagaggtt atttcaggcc atggtgctgc
     5221 gcagatccgc gtatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     5281 aggaaattgt aagcgttaat attttgttaa aattcgattt taccacattt gtagaggttt
     5341 tacttgctta aaaaaacctc ccacatctcc ccctgaacct gaaacataaa atgaatgcaa
     5401 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5461 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5521 tcaatgtatc ttatcatgtc tgctcgaagc ggccgcccca aggggttatg ctagttattg
     5581 ctcagcggtg gcagcagcca actcagcttc ctttcgggct ttgttagcag ccggatcagc
     5641 ttgcatgcct gcaggtcgac tctagaatgc ttgcctatgt ggaaggagag gttttttgtt
     5701 atttccttgg ggtcttgcta agttttccag gctgcactag aactcgtggg agcaatcctt
     5761 ctgcctcagc ctcccaagta gctgggactc cagggagaga ttggtgtttg cacggccata
     5821 ggtagatagc ttatttttac tgcttgccaa caaaattgat cctgaagccc aacggcctta
     5881 tgccaaatgg ttccctcccc actgctgtcc atcctgggct gccactccca aaactgccag
     5941 taccaggtgt cactgattga ctagtcaaat tcctggacaa aggggctaca cactccaggc
     6001 tggttgtgga acttttgttt tgagatggag ttttgctcgt cacccaggct ggagtgcagt
     6061 tgtgcgttct cagctcactg ccacctctgc cttcctggga tcaagcgatt ctcctgcctc
     6121 agccgatatc gtagctggga ttacaggcat acaccaccaa gcctggttaa tttttgtatt
     6181 tttagtagag acggagtttc tccatgttgg ccaggctggt cttgaactcc tgacctcaga
     6241 tgatccaccc acctcaccga attcagtgct gctagcgagc tcgtttaaac aactagaatt
     6301 acacggcgat cttgccgccc ttcttggcct taatgagaat ctcgcggatc ttgcgggcgt
     6361 ccaacttgcc ggtcagtcct ttaggcacct cgtccacgaa cacaacacca ccgcgcagct
     6421 tcttggcggt tgtaacctgg ctggccacat agtccacgat ctccttctcg gtcatggttt
     6481 taccgtgttc cagcacgacg actgcggcgg gcagctcgcc ggcatcgtcg tcgggcaggc
     6541 cggcgacccc ggcgtcgaag atgttggggt gttgcagcag gatgctctcc agttcggctg
     6601 gggctacctg gtagcccttg tatttgatca ggctcttcag ccggtccacg atgaagaagt
     6661 gctcgtcctc gtcccagtag gcgatgtcgc cgctgtgcag ccagccgtcc ttgtcgatga
     6721 gagcgtttgt agcctcgggg ttgttaacgt agccgctcat gatcatgggg ccacggacgc
     6781 acagctcgcc gcgctggttc acacccagtg tcttaccggt gtccaagtcc accaccttag
     6841 cctcgaagaa gggcaccacc ttgcctactg cgccaggctt gtcgtcccct tcgggggtga
     6901 tcagaatggc gctggttgtt tctgtcaggc cgtagccctg gcggatgcct ggtaggtgga
     6961 agcgtttggc cacggcctca cctacctcct tgctgagcgg cgccccgccg ctggcgatct
     7021 cgtgcaagtt gcttaggtcg tacttgtcga tgagagtgct cttagcgaag aagctaaata
     7081 gtgtgggcac cagcagggca gattgaatct tatagtcttg caagctgcgc aagaatagct
     7141 cctcctcgaa gcggtacatg agcacgaccc gaaagccgca gatcaagtag cccagcgtgg
     7201 tgaacatgcc gaagccgtgg tgaaatggca ccacgctgag gatagcggtg tcggggatga
     7261 tctggttgcc gaagatgggg tcgcgggcat gactgaatcg gacacaagcg gtgcggtgcg
     7321 gtagggctac gcccttgggc aatccggtac tgccactact gttcatgatc agggcgatgg
     7381 ttttgtcccg gtcgaagctc tcgggcacga agtcgtactc gttgaagccg ggtggcaaat
     7441 gggaagtcac gaaggtgtac atgctttgga agccctggta gtcggtcttg ctatccatga
     7501 tgatgatctt ttgtatgatc ggtagcttct tttgcacgtt gaggatcttt tgcagccctt
     7561 tcttgctcac gaatacgacg gtgggctggc tgatgcccat gctgttcagc agctcgcgct
     7621 cgttgtagat gtcgttagct ggggccacag ccacaccgat gaacagggca cccaacacgg
     7681 gcatgaagaa ctgcaagcta ttctcgctgc acaccacgat ccgatggttt gtattcagcc
     7741 catagcgctt catagcttct gccagccgaa cgctcatctc gaagtactcg gcgtaggtaa
     7801 tgtccacctc gatatgtgcg tcggtaaagg cgatggtgcc gggcaccagg gcgtagcgct
     7861 tcatggcttt gtgcagctgc tcgccggcgg tcccgtcttc gagtgggtag aatggcgctg
     7921 ggcccttctt aatgtttttg gcatcttc
//
