LOCUS       sequence-409614-        7081 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter plasmid carrying PHLPP2 3'UTR with mutated miR-27b target 
            sequences.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7081)
  AUTHORS   Sakai E, Imaizumi T, Suzuki R, Taracena-Gandara M, Fujimoto T, 
            Sakurai F, Mizuguchi H
  TITLE     miR-27b targets MAIP1 to mediate lipid accumulation in cultured 
            human and mouse hepatic cells.
  JOURNAL   Commun Biol. 2023 Jun 24;6(1):669. doi: 10.1038/s42003-023-05049-w.
  PUBMED    37355744
REFERENCE   2  (bases 1 to 7081)
  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: "Commun 
            Biol."; date: "2023-06-24"; volume: "6(1):669. doi"; pages: " 
            10.1038/s42003-023-05049-w"
COMMENT     Sequence Label: psicheck2-PHLPP2mut
FEATURES             Location/Qualifiers
     source          1..7081
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..419
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        62..253
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(85..105)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      270..405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          489..621
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     604..623
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        666..684
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      688..697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             694..1629
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2496..2544
                     /note="synthetic polyadenylation signal"
     promoter        2553..3304
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      3334..3343
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3340..4992
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /db_xref="GI:42718124"
                     /protein_id="AAS38485.1"
                     /translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     RBS             4299..4307
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    5036..5157
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5047..5066)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5101..5120
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5290..5394
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5395..5463,5464..6255)
                     /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(5613..5632)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6426..7014
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6915..6934
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
       61 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
      121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
      181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
      301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
      361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
      421 agcttgattc ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca
      481 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
      541 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
      601 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga
      661 gtacttaata cgactcacta taggctagcc accatggctt ccaaggtgta cgaccccgag
      721 caacgcaaac gcatgatcac tgggcctcag tggtgggctc gctgcaagca aatgaacgtg
      781 ctggactcct tcatcaacta ctatgattcc gagaagcacg ccgagaacgc cgtgattttt
      841 ctgcatggta acgctgcctc cagctacctg tggaggcacg tcgtgcctca catcgagccc
      901 gtggctagat gcatcatccc tgatctgatc ggaatgggta agtccggcaa gagcgggaat
      961 ggctcatatc gcctcctgga tcactacaag tacctcaccg cttggttcga gctgctgaac
     1021 cttccaaaga aaatcatctt tgtgggccac gactgggggg cttgtctggc ctttcactac
     1081 tcctacgagc accaagacaa gatcaaggcc atcgtccatg ctgagagtgt cgtggacgtg
     1141 atcgagtcct gggacgagtg gcctgacatc gaggaggata tcgccctgat caagagcgaa
     1201 gagggcgaga aaatggtgct tgagaataac ttcttcgtcg agaccatgct cccaagcaag
     1261 atcatgcgga aactggagcc tgaggagttc gctgcctacc tggagccatt caaggagaag
     1321 ggcgaggtta gacggcctac cctctcctgg cctcgcgaga tccctctcgt taagggaggc
     1381 aagcccgacg tcgtccagat tgtccgcaac tacaacgcct accttcgggc cagcgacgat
     1441 ctgcctaaga tgttcatcga gtccgaccct gggttctttt ccaacgctat tgtcgaggga
     1501 gctaagaagt tccctaacac cgagttcgtg aaggtgaagg gcctccactt cagccaggag
     1561 gacgctccag atgaaatggg taagtacatc aagagcttcg tggagcgcgt gctgaagaac
     1621 gagcagtaat tctaggcgat cgctcgagca gtcctaccac ctgccattaa ccctttctct
     1681 cctaggatca ttttgagaat ttgcctgcct gggcaggaaa gggactattt ctgtggagga
     1741 aaaaagtgaa gattgattct ctttactagt tgctgctgat ggatctctgt gacagagaaa
     1801 tcaccttatc tcagactaat ggggtgtgat gtgactagtc acatggcttt tcattcttct
     1861 ctacgagaat acagcctatc aaaatgatgt ctgttggaaa tgtagaacca atcaaacaga
     1921 taatttatgt atgtaatgta atgagagcac ttttcattga ttataaactt tttatttttg
     1981 aatctgcact cgagccaatc ttcttagagg cagcccggca ccttcatcca taggcagaga
     2041 gagaactggg tgttggagac ttattcgagg gtataggaag ggccttataa agttgattta
     2101 acttttggat gtcagattat aaaagctcct gagaaacttg gggtaatagg atcttctttt
     2161 ggggatgaaa atggggaagg cgtgaggacc tagactactt ctccctagat cagaaaaaga
     2221 gaattacccc ttgacaaata tgatacctgc taggtatttc ccagggaaat ttagggattg
     2281 gcctctttcc ctagcatgtg gaggaattgg cagacagctt cctaagggcg gggagcgggg
     2341 gcccaaggcc gacactgctt gcatccacgt gaccttaagt tatggcagat gactctgaaa
     2401 cggactgagg ccaatgagaa cagatggatg gagcactcag gttagacttg ttccttctcc
     2461 tatgctggag gagagggatg gcggccgctg gccgcaataa aatatcttta ttttcattac
     2521 atctgtgtgt tggttttttg tgtgaggatc taaatgagtc ttcggacctc gcgggggccg
     2581 cttaagcggt ggttagggtt tgtctgacgc ggggggaggg ggaaggaacg aaacactctc
     2641 attcggaggc ggctcggggt ttggtcttgg tggccacggg cacgcagaag agcgccgcga
     2701 tcctcttaag cacccccccg ccctccgtgg aggcgggggt ttggtcggcg ggtggtaact
     2761 ggcgggccgc tgactcgggc gggtcgcgcg ccccagagtg tgaccttttc ggtctgctcg
     2821 cagacccccg ggcggcgccg ccgcggcggc gacgggctcg ctgggtccta ggctccatgg
     2881 ggaccgtata cgtggacagg ctctggagca tccgcacgac tgcggtgata ttaccggaga
     2941 ccttctgcgg gacgagccgg gtcacgcggc tgacgcggag cgtccgttgg gcgacaaaca
     3001 ccaggacggg gcacaggtac actatcttgt cacccggagg cgcgagggac tgcaggagct
     3061 tcagggagtg gcgcagctgc ttcatccccg tggcccgttg ctcgcgtttg ctggcggtgt
     3121 ccccggaaga aatatatttg catgtcttta gttctatgat gacacaaacc ccgcccagcg
     3181 tcttgtcatt ggcgaattcg aacacgcaga tgcagtcggg gcggcgcggt cccaggtcca
     3241 cttcgcatat taaggtgacg cgtgtggcct cgaacaccga gcgaccctgc agcgacccgc
     3301 ttaaaagctt ggcattccgg tactgttggt aaagccacca tggccgatgc taagaacatt
     3361 aagaagggcc ctgctccctt ctaccctctg gaggatggca ccgctggcga gcagctgcac
     3421 aaggccatga agaggtatgc cctggtgcct ggcaccattg ccttcaccga tgcccacatt
     3481 gaggtggaca tcacctatgc cgagtacttc gagatgtctg tgcgcctggc cgaggccatg
     3541 aagaggtacg gcctgaacac caaccaccgc atcgtggtgt gctctgagaa ctctctgcag
     3601 ttcttcatgc cagtgctggg cgccctgttc atcggagtgg ccgtggcccc tgctaacgac
     3661 atttacaacg agcgcgagct gctgaacagc atgggcattt ctcagcctac cgtggtgttc
     3721 gtgtctaaga agggcctgca gaagatcctg aacgtgcaga agaagctgcc tatcatccag
     3781 aagatcatca tcatggactc taagaccgac taccagggct tccagagcat gtacacattc
     3841 gtgacatctc atctgcctcc tggcttcaac gagtacgact tcgtgccaga gtctttcgac
     3901 agggacaaaa ccattgccct gatcatgaac agctctgggt ctaccggcct gcctaagggc
     3961 gtggccctgc ctcatcgcac cgcctgtgtg cgcttctctc acgcccgcga ccctattttc
     4021 ggcaaccaga tcatccccga caccgctatt ctgagcgtgg tgccattcca ccacggcttc
     4081 ggcatgttca ccaccctggg ctacctgatt tgcggctttc gggtggtgct gatgtaccgc
     4141 ttcgaggagg agctgttcct gcgcagcctg caagactaca aaattcagtc tgccctgctg
     4201 gtgccaaccc tgttcagctt cttcgctaag agcaccctga tcgacaagta cgacctgtct
     4261 aacctgcacg agattgcctc tggcggcgcc ccactgtcta aggaggtggg cgaagccgtg
     4321 gccaagcgct ttcatctgcc aggcatccgc cagggctacg gcctgaccga gacaaccagc
     4381 gccattctga ttaccccaga gggcgacgac aagcctggcg ccgtgggcaa ggtggtgcca
     4441 ttcttcgagg ccaaggtggt ggacctggac accggcaaga ccctgggagt gaaccagcgc
     4501 ggcgagctgt gtgtgcgcgg ccctatgatt atgtccggct acgtgaataa ccctgaggcc
     4561 acaaacgccc tgatcgacaa ggacggctgg ctgcactctg gcgacattgc ctactgggac
     4621 gaggacgagc acttcttcat cgtggaccgc ctgaagtctc tgatcaagta caagggctac
     4681 caggtggccc cagccgagct ggagtctatc ctgctgcagc accctaacat tttcgacgcc
     4741 ggagtggccg gcctgcccga cgacgatgcc ggcgagctgc ctgccgccgt cgtcgtgctg
     4801 gaacacggca agaccatgac cgagaaggag atcgtggact atgtggccag ccaggtgaca
     4861 accgccaaga agctgcgcgg cggagtggtg ttcgtggacg aggtgcccaa gggcctgacc
     4921 ggcaagctgg acgcccgcaa gatccgcgag atcctgatca aggctaagaa aggcggcaag
     4981 atcgccgtgt aataattcta gagtcggggc ggccggccgc ttcgagcaga catgataaga
     5041 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     5101 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     5161 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     5221 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atccaggtgg cacttttcgg
     5281 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5341 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5401 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5461 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5521 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5581 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5641 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5701 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5761 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5821 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5881 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5941 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     6001 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6061 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6121 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6181 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6241 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6301 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6361 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6421 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6481 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6541 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6601 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6661 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6721 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6781 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6841 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6901 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6961 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     7021 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catggctcga
     7081 c
//
