LOCUS       sequence-359483-        7948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human PDAP1 3` UTR region mutated in 4x 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-Mut4x
FEATURES             Location/Qualifiers
     source          1..7948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..534
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(70..93)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     467..486
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      575..584
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             581..2233
                     /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    2999..3120
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3010..3029)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3064..3083
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3365..3722
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3365..3556
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3388..3408)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3573..3708
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3767..3776
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3773..4705
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLRDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             4757..5548
                     /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(4808..4827)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5418..5437
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5612..5660
                     /note="synthetic polyadenylation signal"
     promoter        5816..5920
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5921..5989,5990..6781)
                     /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(6139..6158)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6939..7527)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7019..7038)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7643..7926
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 acgatgcgtc cggcgtagag gatcgagatc taccgggtag gggaggcgct tttcccaagg
       61 cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc
      121 tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc
      181 cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc
      241 gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt gcagatggac
      301 agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc
      361 tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg gctcaggggc
      421 gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt ctgcacgctt
      481 caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct ttcgacctgc
      541 agcccaagct tggcaatccg gtactgttgg taaagccacc atggaagatg ccaaaaacat
      601 taagaagggc ccagcgccat tctacccact cgaagacggg accgccggcg agcagctgca
      661 caaagccatg aagcgctacg ccctggtgcc cggcaccatc gcctttaccg acgcacatat
      721 cgaggtggac attacctacg ccgagtactt cgagatgagc gttcggctgg cagaagctat
      781 gaagcgctat gggctgaata caaaccatcg gatcgtggtg tgcagcgaga atagcttgca
      841 gttcttcatg cccgtgttgg gtgccctgtt catcggtgtg gctgtggccc cagctaacga
      901 catctacaac gagcgcgagc tgctgaacag catgggcatc agccagccca ccgtcgtatt
      961 cgtgagcaag aaagggctgc aaaagatcct caacgtgcaa aagaagctac cgatcataca
     1021 aaagatcatc atcatggata gcaagaccga ctaccagggc ttccaaagca tgtacacctt
     1081 cgtgacttcc catttgccac ccggcttcaa cgagtacgac ttcgtgcccg agagcttcga
     1141 ccgggacaaa accatcgccc tgatcatgaa cagtagtggc agtaccggat tgcccaaggg
     1201 cgtagcccta ccgcaccgca ccgcttgtgt ccgattcagt catgcccgcg accccatctt
     1261 cggcaaccag atcatccccg acaccgctat cctcagcgtg gtgccatttc accacggctt
     1321 cggcatgttc accacgctgg gctacttgat ctgcggcttt cgggtcgtgc tcatgtaccg
     1381 cttcgaggag gagctattct tgcgcagctt gcaagactat aagattcaat ctgccctgct
     1441 ggtgcccaca ctatttagct tcttcgctaa gagcactctc atcgacaagt acgacctaag
     1501 caacttgcac gagatcgcca gcggcggggc gccgctcagc aaggaggtag gtgaggccgt
     1561 ggccaaacgc ttccacctac caggcatccg ccagggctac ggcctgacag aaacaaccag
     1621 cgccattctg atcacccccg aaggggacga caagcctggc gcagtaggca aggtggtgcc
     1681 cttcttcgag gctaaggtgg tggacttgga caccggtaag acactgggtg tgaaccagcg
     1741 cggcgagctg tgcgtccgtg gccccatgat catgagcggc tacgttaaca accccgaggc
     1801 tacaaacgct ctcatcgaca aggacggctg gctgcacagc ggcgacatcg cctactggga
     1861 cgaggacgag cacttcttca tcgtggaccg gctgaagagc ctgatcaaat acaagggcta
     1921 ccaggtagcc ccagccgaac tggagagcat cctgctgcaa caccccaaca tcttcgacgc
     1981 cggggtcgcc ggcctgcccg acgacgatgc cggcgagctg cccgccgcag tcgtcgtgct
     2041 ggaacacggt aaaaccatga ccgagaagga gatcgtggac tatgtggcca gccaggttac
     2101 aaccgccaag aagctgcgcg gtggtgttgt gttcgtggac gaggtgccta aaggactgac
     2161 cggcaagttg gacgcccgca agatccgcga gattctcatt aaggccaaga agggcggcaa
     2221 gatcgccgtg taattctagt tgtttaaacg agctcgctag cagcactgaa ttcggtgagg
     2281 tgggtggatc atctgaggtc aggagttcaa gaccagcctg gccaacatgg agaaactccg
     2341 tctctactaa aaatacaaaa attaaccagg cttggtggtg tatgcctgta atcccagcta
     2401 cgatatcggc tgaggcagga gaatcgcttg atcccaggaa ggcagaggtg gcagtgagct
     2461 gagaacgcac aactgcactc cagcctgggt gacgagcaaa actccatctc aaaacaaaag
     2521 ttccacaacc agcctggagt gtgtagcccc tttgtccagg aatttgacta gtcaatcagt
     2581 gacacctggt actggcagtt gatatcgtgg cagcccagga tggacagcag tggggaggga
     2641 accatttggc ataaggccgt tgggcttcag gatcaatttt gttggcaagc agtaaaaata
     2701 agctatctac ctatggccgt gcaaacacca atctctccct ggagtcccag ctacgaattc
     2761 ggctgaggca gaaggattgc tcccacgagt tctagtgcag cctggaaaac ttagcaagac
     2821 cccaaggaaa taacaaaaaa cctctccttc cacataggca agcattctag agtcgacctg
     2881 caggcatgca agctgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
     2941 ccaccgctga gcaataacta gcataacccc ttggggcggc cgcttcgagc agacatgata
     3001 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     3061 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     3121 aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt
     3181 ttaagcaagt aaaacctcta caaatgtggt aaaatcgaat tttaacaaaa tattaacgct
     3241 tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     3301 cgcggatctg cgcagcacca tggcctgaaa taacctctga aagaggaact tggttaggta
     3361 ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3421 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3481 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3541 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3601 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3661 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3721 aaaagcttga ttcttctgac acaacagtct cgaaccaaag gctggagcca ccatggcttc
     3781 caaggtgtac gaccccgagc aacgcaaacg catgatcact gggcctcagt ggtgggctcg
     3841 ctgcaagcaa atgaacgtgc tggactcctt catcaactac tatgattccg agaagcacgc
     3901 cgagaacgcc gtgatttttc tgcatggtaa cgctgcctcc agctacctgt ggaggcacgt
     3961 cgtgcctcac atcgagcccg tggctagatg catcatccct gatctgatcg gaatgggtaa
     4021 gtccggcaag agcgggaatg gctcatatcg cctccgggat cactacaagt acctcaccgc
     4081 ttggttcgag ctgctgaacc ttccaaagaa aatcatcttt gtgggccacg actggggggc
     4141 ttgtctggcc tttcactact cctacgagca ccaagacaag atcaaggcca tcgtccatgc
     4201 tgagagtgtc gtggacgtga tcgagtcctg ggacgagtgg cctgacatcg aggaggatat
     4261 cgccctgatc aagagcgaag agggcgagaa aatggtgctt gagaataact tcttcgtcga
     4321 gaccatgctc ccaagcaaga tcatgcggaa actggagcct gaggagttcg ctgcctacct
     4381 ggagccattc aaggagaagg gcgaggttag acggcctacc ctctcctggc ctcgcgagat
     4441 ccctctcgtt aagggaggca agcccgacgt cgtccagatt gtccgcaact acaacgccta
     4501 ccttcgggcc agcgacgatc tgcctaagat gttcatcgag tccgaccctg ggttcttttc
     4561 caacgctatt gtcgagggag ctaagaagtt ccctaacacc gagttcgtga aggtgaaggg
     4621 cctccacttc agccaggagg acgctccaga tgaaatgggt aagtacatca agagcttcgt
     4681 ggagcgcgtg ctgaagaacg agcagaccgg tggtgggagc ggaggtggcg gatcaggtgg
     4741 cggaggctcc ggagggattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4801 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4861 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4921 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4981 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     5041 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     5101 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     5161 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     5221 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     5281 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5341 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5401 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5461 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5521 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     5581 gcgacgccca acctgccatc acgatggccg caataaaata tctttatttt cattacatct
     5641 gtgtgttggt tttttgtgtg aatcgatagc gataaggatc ctctttgcgc ttgcgttttc
     5701 ccttgtccag atagcccagt agctgacatt catccggggt cagcaccgtt tctgcggact
     5761 ggctttctac gtaatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     5821 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5881 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5941 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     6001 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     6061 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     6121 gagcactttc aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     6181 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     6241 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     6301 gagtgataac actgcggcca acttacttct gacaactatc ggaggaccga aggagctaac
     6361 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6421 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6481 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6541 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6601 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6661 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6721 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6781 attcgaaatg accgaccaag cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata
     6841 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6901 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6961 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     7021 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7081 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     7141 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7201 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7261 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7321 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     7381 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7441 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     7501 attacgcgca gaaaaaaagg atttcaagaa gatcctttga tcttttctac ggggtctgac
     7561 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7621 ttcacctaga tccttttata gtccggaaat acaggaacgc acgctggatg gcccttcgct
     7681 gggatggtga aaccatgaaa aatggcagct tcagtggatt aagtgggggt aatgtggcct
     7741 gtaccctctg gttgcatagg tattcatacg gttaaaattt atcaggcgcg attgcggcag
     7801 tttttcgggt ggtttgttgc catttttacc tgtctgctgc cgtgatcgcg ctgaacgcgt
     7861 tttagcggtg cgtacaatta agggattatg gtaaatccac ttactgtctg ccctcgtagc
     7921 catcgagata aaccgcagta ctccggcc
//
