LOCUS       sequence-409617-        6291 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter plasmid carrying miR-27b target sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6291)
  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 6291)
  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: psicheck2-27bT
FEATURES             Location/Qualifiers
     source          1..6291
                     /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    1706..1754
                     /note="synthetic polyadenylation signal"
     promoter        1763..2514
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      2544..2553
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2550..4202
                     /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             3509..3517
                     /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    4246..4367
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4257..4276)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4311..4330
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4500..4604
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4605..4673,4674..5465)
                     /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(4823..4842)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5636..6224
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6125..6144
                     /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 cgctcgaggc agaacttagc cactgtgaag cagaacttag
     1681 ccactgtgaa gcggccgctg gccgcaataa aatatcttta ttttcattac atctgtgtgt
     1741 tggttttttg tgtgaggatc taaatgagtc ttcggacctc gcgggggccg cttaagcggt
     1801 ggttagggtt tgtctgacgc ggggggaggg ggaaggaacg aaacactctc attcggaggc
     1861 ggctcggggt ttggtcttgg tggccacggg cacgcagaag agcgccgcga tcctcttaag
     1921 cacccccccg ccctccgtgg aggcgggggt ttggtcggcg ggtggtaact ggcgggccgc
     1981 tgactcgggc gggtcgcgcg ccccagagtg tgaccttttc ggtctgctcg cagacccccg
     2041 ggcggcgccg ccgcggcggc gacgggctcg ctgggtccta ggctccatgg ggaccgtata
     2101 cgtggacagg ctctggagca tccgcacgac tgcggtgata ttaccggaga ccttctgcgg
     2161 gacgagccgg gtcacgcggc tgacgcggag cgtccgttgg gcgacaaaca ccaggacggg
     2221 gcacaggtac actatcttgt cacccggagg cgcgagggac tgcaggagct tcagggagtg
     2281 gcgcagctgc ttcatccccg tggcccgttg ctcgcgtttg ctggcggtgt ccccggaaga
     2341 aatatatttg catgtcttta gttctatgat gacacaaacc ccgcccagcg tcttgtcatt
     2401 ggcgaattcg aacacgcaga tgcagtcggg gcggcgcggt cccaggtcca cttcgcatat
     2461 taaggtgacg cgtgtggcct cgaacaccga gcgaccctgc agcgacccgc ttaaaagctt
     2521 ggcattccgg tactgttggt aaagccacca tggccgatgc taagaacatt aagaagggcc
     2581 ctgctccctt ctaccctctg gaggatggca ccgctggcga gcagctgcac aaggccatga
     2641 agaggtatgc cctggtgcct ggcaccattg ccttcaccga tgcccacatt gaggtggaca
     2701 tcacctatgc cgagtacttc gagatgtctg tgcgcctggc cgaggccatg aagaggtacg
     2761 gcctgaacac caaccaccgc atcgtggtgt gctctgagaa ctctctgcag ttcttcatgc
     2821 cagtgctggg cgccctgttc atcggagtgg ccgtggcccc tgctaacgac atttacaacg
     2881 agcgcgagct gctgaacagc atgggcattt ctcagcctac cgtggtgttc gtgtctaaga
     2941 agggcctgca gaagatcctg aacgtgcaga agaagctgcc tatcatccag aagatcatca
     3001 tcatggactc taagaccgac taccagggct tccagagcat gtacacattc gtgacatctc
     3061 atctgcctcc tggcttcaac gagtacgact tcgtgccaga gtctttcgac agggacaaaa
     3121 ccattgccct gatcatgaac agctctgggt ctaccggcct gcctaagggc gtggccctgc
     3181 ctcatcgcac cgcctgtgtg cgcttctctc acgcccgcga ccctattttc ggcaaccaga
     3241 tcatccccga caccgctatt ctgagcgtgg tgccattcca ccacggcttc ggcatgttca
     3301 ccaccctggg ctacctgatt tgcggctttc gggtggtgct gatgtaccgc ttcgaggagg
     3361 agctgttcct gcgcagcctg caagactaca aaattcagtc tgccctgctg gtgccaaccc
     3421 tgttcagctt cttcgctaag agcaccctga tcgacaagta cgacctgtct aacctgcacg
     3481 agattgcctc tggcggcgcc ccactgtcta aggaggtggg cgaagccgtg gccaagcgct
     3541 ttcatctgcc aggcatccgc cagggctacg gcctgaccga gacaaccagc gccattctga
     3601 ttaccccaga gggcgacgac aagcctggcg ccgtgggcaa ggtggtgcca ttcttcgagg
     3661 ccaaggtggt ggacctggac accggcaaga ccctgggagt gaaccagcgc ggcgagctgt
     3721 gtgtgcgcgg ccctatgatt atgtccggct acgtgaataa ccctgaggcc acaaacgccc
     3781 tgatcgacaa ggacggctgg ctgcactctg gcgacattgc ctactgggac gaggacgagc
     3841 acttcttcat cgtggaccgc ctgaagtctc tgatcaagta caagggctac caggtggccc
     3901 cagccgagct ggagtctatc ctgctgcagc accctaacat tttcgacgcc ggagtggccg
     3961 gcctgcccga cgacgatgcc ggcgagctgc ctgccgccgt cgtcgtgctg gaacacggca
     4021 agaccatgac cgagaaggag atcgtggact atgtggccag ccaggtgaca accgccaaga
     4081 agctgcgcgg cggagtggtg ttcgtggacg aggtgcccaa gggcctgacc ggcaagctgg
     4141 acgcccgcaa gatccgcgag atcctgatca aggctaagaa aggcggcaag atcgccgtgt
     4201 aataattcta gagtcggggc ggccggccgc ttcgagcaga catgataaga tacattgatg
     4261 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     4321 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     4381 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     4441 acctctacaa atgtggtaaa atcgataagg atccaggtgg cacttttcgg ggaaatgtgc
     4501 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4561 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4621 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4681 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4741 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4801 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     4861 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     4921 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     4981 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5041 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5101 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5161 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5221 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5281 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5341 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5401 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5461 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5521 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5581 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5641 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5701 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5761 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5821 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5881 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5941 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6001 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6061 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6121 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6181 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6241 cctttttacg gttcctggcc ttttgctggc cttttgctca catggctcga c
//
