LOCUS       sequence-357586-        7877 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human MYB 3` UTR region containing 3x 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 7877)
  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 7877)
  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-MYB-3'UTR
FEATURES             Location/Qualifiers
     source          1..7877
                     /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    2416..2537
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2427..2446)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2481..2500
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2782..3139
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2782..2973
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2805..2825)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2990..3125
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3184..3193
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3190..4122
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             4174..4965
                     /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(4225..4244)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4835..4854
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5029..5077
                     /note="synthetic polyadenylation signal"
     promoter        5233..5337
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5338..5406,5407..6198)
                     /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(5556..5575)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6356..6944)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6436..6455)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7060..7343
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     promoter        join(7400..7877,1..22)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7435..7458)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     7832..7851
                     /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 ctcgctagct gtaaatgcca aatgaaaaac gttttttgct gctatggtct tagcctgtag
     1801 acatgctgct agtatcagag gggcagtaga gcttggacag aaagaaaaga aacttggtgt
     1861 taggtaattg actatgcact agtatttcag actttttaat tttatatata tatacatttt
     1921 ttttccttct gcaatacatt tgaaaacttg tttgggagac tctgcatttt ttattgtggt
     1981 ttttttgtta ttgttggttt atacaagcat gcgttgcact tcttttttgg gagatgtgtg
     2041 ttgttgatgt tctatgtttt gttttgagtg tagcctgact gttttataat ttgggagttc
     2101 tgcatttgat ccgcatcccc tgtggtttct aagtgtatgg tctcagaact gttgcatgga
     2161 tcctgtgttt gcaactgggg agacagaaac tgtggttgat agccagtcac tgccttaaga
     2221 acatttgatg caagatggcc agcactgaac ttttgagata tgacggtgta cttactgcct
     2281 tgtctagagt cgacctgcag gcatgcaagc tgatccggct gctaacaaag cccgaaagga
     2341 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcggccgc
     2401 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     2461 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     2521 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     2581 agatgtggga ggttttttta agcaagtaaa acctctacaa atgtggtaaa atcgaatttt
     2641 aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg
     2701 gtatttcaca ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag
     2761 aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     2821 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2881 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2941 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3001 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3061 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3121 aggcctaggc ttttgcaaaa agcttgattc ttctgacaca acagtctcga accaaaggct
     3181 ggagccacca tggcttccaa ggtgtacgac cccgagcaac gcaaacgcat gatcactggg
     3241 cctcagtggt gggctcgctg caagcaaatg aacgtgctgg actccttcat caactactat
     3301 gattccgaga agcacgccga gaacgccgtg atttttctgc atggtaacgc tgcctccagc
     3361 tacctgtgga ggcacgtcgt gcctcacatc gagcccgtgg ctagatgcat catccctgat
     3421 ctgatcggaa tgggtaagtc cggcaagagc gggaatggct catatcgcct cctggatcac
     3481 tacaagtacc tcaccgcttg gttcgagctg ctgaaccttc caaagaaaat catctttgtg
     3541 ggccacgact ggggggcttg tctggccttt cactactcct acgagcacca agacaagatc
     3601 aaggccatcg tccatgctga gagtgtcgtg gacgtgatcg agtcctggga cgagtggcct
     3661 gacatcgagg aggatatcgc cctgatcaag agcgaagagg gcgagaaaat ggtgcttgag
     3721 aataacttct tcgtcgagac catgctccca agcaagatca tgcggaaact ggagcctgag
     3781 gagttcgctg cctacctgga gccattcaag gagaagggcg aggttagacg gcctaccctc
     3841 tcctggcctc gcgagatccc tctcgttaag ggaggcaagc ccgacgtcgt ccagattgtc
     3901 cgcaactaca acgcctacct tcgggccagc gacgatctgc ctaagatgtt catcgagtcc
     3961 gaccctgggt tcttttccaa cgctattgtc gagggagcta agaagttccc taacaccgag
     4021 ttcgtgaagg tgaagggcct ccacttcagc caggaggacg ctccagatga aatgggtaag
     4081 tacatcaaga gcttcgtgga gcgcgtgctg aagaacgagc agaccggtgg tgggagcgga
     4141 ggtggcggat caggtggcgg aggctccgga gggattgaac aagatggatt gcacgcaggt
     4201 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4261 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4321 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     4381 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4441 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4501 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4561 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4621 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4681 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4741 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4801 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4861 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4921 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4981 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg atggccgcaa taaaatatct
     5041 ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagcgat aaggatcctc
     5101 tttgcgcttg cgttttccct tgtccagata gcccagtagc tgacattcat ccggggtcag
     5161 caccgtttct gcggactggc tttctacgta atggtttctt agacgtcagg tggcactttt
     5221 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5281 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5341 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5401 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5461 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5521 gaacgttttc caatgatgag cactttcaaa gttctgctat gtggcgcggt attatcccgt
     5581 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5641 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5701 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aactatcgga
     5761 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5821 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5881 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5941 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6001 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6061 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6121 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6181 ctgattaagc attggtaatt cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc
     6241 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6301 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6361 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6421 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6481 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6541 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6601 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6661 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6721 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6781 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6841 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6901 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat cctttgatct
     6961 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     7021 gattatcaaa aaggatcttc acctagatcc ttttatagtc cggaaataca ggaacgcacg
     7081 ctggatggcc cttcgctggg atggtgaaac catgaaaaat ggcagcttca gtggattaag
     7141 tgggggtaat gtggcctgta ccctctggtt gcataggtat tcatacggtt aaaatttatc
     7201 aggcgcgatt gcggcagttt ttcgggtggt ttgttgccat ttttacctgt ctgctgccgt
     7261 gatcgcgctg aacgcgtttt agcggtgcgt acaattaagg gattatggta aatccactta
     7321 ctgtctgccc tcgtagccat cgagataaac cgcagtactc cggccacgat gcgtccggcg
     7381 tagaggatcg agatctaccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc
     7441 gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat
     7501 tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct
     7561 tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg
     7621 tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa
     7681 tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg
     7741 gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg
     7801 gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct
     7861 gccgcgctgt tctcctc
//
