LOCUS       sequence-409616-        6420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter plasmid carrying MAIP1 3'UTR with mutated miR27b target 
            sequences.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6420)
  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 6420)
  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-MAIP1mut
FEATURES             Location/Qualifiers
     source          1..6420
                     /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    1835..1883
                     /note="synthetic polyadenylation signal"
     promoter        1892..2643
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      2673..2682
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2679..4331
                     /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             3638..3646
                     /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    4375..4496
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4386..4405)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4440..4459
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4629..4733
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4734..4802,4803..5594)
                     /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(4952..4971)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5765..6353
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6254..6273
                     /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 cgctcgagtc agcttatgga ctttagcagt tgttataaaa
     1681 aactaaggaa gaaaaatttt ggggtcattt gatcttcact taatctaagt ttataaatta
     1741 cttttatatt attttgaaat actccttgca gtatattggc atgatacagt aaaagcattt
     1801 tccacagatt gttatcaccg cggccgctgg ccgcaataaa atatctttat tttcattaca
     1861 tctgtgtgtt ggttttttgt gtgaggatct aaatgagtct tcggacctcg cgggggccgc
     1921 ttaagcggtg gttagggttt gtctgacgcg gggggagggg gaaggaacga aacactctca
     1981 ttcggaggcg gctcggggtt tggtcttggt ggccacgggc acgcagaaga gcgccgcgat
     2041 cctcttaagc acccccccgc cctccgtgga ggcgggggtt tggtcggcgg gtggtaactg
     2101 gcgggccgct gactcgggcg ggtcgcgcgc cccagagtgt gaccttttcg gtctgctcgc
     2161 agacccccgg gcggcgccgc cgcggcggcg acgggctcgc tgggtcctag gctccatggg
     2221 gaccgtatac gtggacaggc tctggagcat ccgcacgact gcggtgatat taccggagac
     2281 cttctgcggg acgagccggg tcacgcggct gacgcggagc gtccgttggg cgacaaacac
     2341 caggacgggg cacaggtaca ctatcttgtc acccggaggc gcgagggact gcaggagctt
     2401 cagggagtgg cgcagctgct tcatccccgt ggcccgttgc tcgcgtttgc tggcggtgtc
     2461 cccggaagaa atatatttgc atgtctttag ttctatgatg acacaaaccc cgcccagcgt
     2521 cttgtcattg gcgaattcga acacgcagat gcagtcgggg cggcgcggtc ccaggtccac
     2581 ttcgcatatt aaggtgacgc gtgtggcctc gaacaccgag cgaccctgca gcgacccgct
     2641 taaaagcttg gcattccggt actgttggta aagccaccat ggccgatgct aagaacatta
     2701 agaagggccc tgctcccttc taccctctgg aggatggcac cgctggcgag cagctgcaca
     2761 aggccatgaa gaggtatgcc ctggtgcctg gcaccattgc cttcaccgat gcccacattg
     2821 aggtggacat cacctatgcc gagtacttcg agatgtctgt gcgcctggcc gaggccatga
     2881 agaggtacgg cctgaacacc aaccaccgca tcgtggtgtg ctctgagaac tctctgcagt
     2941 tcttcatgcc agtgctgggc gccctgttca tcggagtggc cgtggcccct gctaacgaca
     3001 tttacaacga gcgcgagctg ctgaacagca tgggcatttc tcagcctacc gtggtgttcg
     3061 tgtctaagaa gggcctgcag aagatcctga acgtgcagaa gaagctgcct atcatccaga
     3121 agatcatcat catggactct aagaccgact accagggctt ccagagcatg tacacattcg
     3181 tgacatctca tctgcctcct ggcttcaacg agtacgactt cgtgccagag tctttcgaca
     3241 gggacaaaac cattgccctg atcatgaaca gctctgggtc taccggcctg cctaagggcg
     3301 tggccctgcc tcatcgcacc gcctgtgtgc gcttctctca cgcccgcgac cctattttcg
     3361 gcaaccagat catccccgac accgctattc tgagcgtggt gccattccac cacggcttcg
     3421 gcatgttcac caccctgggc tacctgattt gcggctttcg ggtggtgctg atgtaccgct
     3481 tcgaggagga gctgttcctg cgcagcctgc aagactacaa aattcagtct gccctgctgg
     3541 tgccaaccct gttcagcttc ttcgctaaga gcaccctgat cgacaagtac gacctgtcta
     3601 acctgcacga gattgcctct ggcggcgccc cactgtctaa ggaggtgggc gaagccgtgg
     3661 ccaagcgctt tcatctgcca ggcatccgcc agggctacgg cctgaccgag acaaccagcg
     3721 ccattctgat taccccagag ggcgacgaca agcctggcgc cgtgggcaag gtggtgccat
     3781 tcttcgaggc caaggtggtg gacctggaca ccggcaagac cctgggagtg aaccagcgcg
     3841 gcgagctgtg tgtgcgcggc cctatgatta tgtccggcta cgtgaataac cctgaggcca
     3901 caaacgccct gatcgacaag gacggctggc tgcactctgg cgacattgcc tactgggacg
     3961 aggacgagca cttcttcatc gtggaccgcc tgaagtctct gatcaagtac aagggctacc
     4021 aggtggcccc agccgagctg gagtctatcc tgctgcagca ccctaacatt ttcgacgccg
     4081 gagtggccgg cctgcccgac gacgatgccg gcgagctgcc tgccgccgtc gtcgtgctgg
     4141 aacacggcaa gaccatgacc gagaaggaga tcgtggacta tgtggccagc caggtgacaa
     4201 ccgccaagaa gctgcgcggc ggagtggtgt tcgtggacga ggtgcccaag ggcctgaccg
     4261 gcaagctgga cgcccgcaag atccgcgaga tcctgatcaa ggctaagaaa ggcggcaaga
     4321 tcgccgtgta ataattctag agtcggggcg gccggccgct tcgagcagac atgataagat
     4381 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     4441 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     4501 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     4561 gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tccaggtggc acttttcggg
     4621 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4681 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4741 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4801 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4861 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4921 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4981 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5041 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5101 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5161 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5221 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5281 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5341 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5401 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5461 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5521 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5581 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5641 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5701 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5761 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5821 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5881 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5941 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6001 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6061 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6121 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6181 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6241 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6301 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6361 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atggctcgac
//
