LOCUS       sequence-409682-        6778 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter plasmid carrying B4GALT3 3'UTR with mutated miR27b target 
            sequences..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6778)
  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 6778)
  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-B4GALT3mut
FEATURES             Location/Qualifiers
     source          1..6778
                     /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    2193..2241
                     /note="synthetic polyadenylation signal"
     promoter        2250..3001
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      3031..3040
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3037..4689
                     /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             3996..4004
                     /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    4733..4854
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4744..4763)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4798..4817
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4987..5091
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5092..5160,5161..5952)
                     /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(5310..5329)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6123..6711
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6612..6631
                     /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 cctggttcct cccaagcctt ccgtcaagag
     1681 atgctgcaac gccggccccc agccaggcct gggcctctat ctactgccaa ccacacagcc
     1741 ctccgaggtt cacactgact cctccttcct gtctacctta atcatgaaac cgaattcatg
     1801 gggttgtatt ctccccaccc tcagctcctc actgttctca gagggatgtg agggaactga
     1861 actctggtgc cgtgctaggg ggtaggggcc tctccctcac tgctggactg gagctgggct
     1921 cctgtagacc tgaggggtcc ctctctctag ggtctcctgt agggcttatg attataaatc
     1981 cttgatgtca tgattttatg tgacgattcc taggagtccc tgcccctaga gtaggagcag
     2041 ggctggaccc caagcccctc cctcttccat ggagagaaga gtgatctggc ttctcctcgg
     2101 acctttataa atatttattc tatttatggt tcccgggaag ttgtttggtg aaggaagccc
     2161 ctccctgggc attttctgcg gccgctggcc gcaataaaat atctttattt tcattacatc
     2221 tgtgtgttgg ttttttgtgt gaggatctaa atgagtcttc ggacctcgcg ggggccgctt
     2281 aagcggtggt tagggtttgt ctgacgcggg gggaggggga aggaacgaaa cactctcatt
     2341 cggaggcggc tcggggtttg gtcttggtgg ccacgggcac gcagaagagc gccgcgatcc
     2401 tcttaagcac ccccccgccc tccgtggagg cgggggtttg gtcggcgggt ggtaactggc
     2461 gggccgctga ctcgggcggg tcgcgcgccc cagagtgtga ccttttcggt ctgctcgcag
     2521 acccccgggc ggcgccgccg cggcggcgac gggctcgctg ggtcctaggc tccatgggga
     2581 ccgtatacgt ggacaggctc tggagcatcc gcacgactgc ggtgatatta ccggagacct
     2641 tctgcgggac gagccgggtc acgcggctga cgcggagcgt ccgttgggcg acaaacacca
     2701 ggacggggca caggtacact atcttgtcac ccggaggcgc gagggactgc aggagcttca
     2761 gggagtggcg cagctgcttc atccccgtgg cccgttgctc gcgtttgctg gcggtgtccc
     2821 cggaagaaat atatttgcat gtctttagtt ctatgatgac acaaaccccg cccagcgtct
     2881 tgtcattggc gaattcgaac acgcagatgc agtcggggcg gcgcggtccc aggtccactt
     2941 cgcatattaa ggtgacgcgt gtggcctcga acaccgagcg accctgcagc gacccgctta
     3001 aaagcttggc attccggtac tgttggtaaa gccaccatgg ccgatgctaa gaacattaag
     3061 aagggccctg ctcccttcta ccctctggag gatggcaccg ctggcgagca gctgcacaag
     3121 gccatgaaga ggtatgccct ggtgcctggc accattgcct tcaccgatgc ccacattgag
     3181 gtggacatca cctatgccga gtacttcgag atgtctgtgc gcctggccga ggccatgaag
     3241 aggtacggcc tgaacaccaa ccaccgcatc gtggtgtgct ctgagaactc tctgcagttc
     3301 ttcatgccag tgctgggcgc cctgttcatc ggagtggccg tggcccctgc taacgacatt
     3361 tacaacgagc gcgagctgct gaacagcatg ggcatttctc agcctaccgt ggtgttcgtg
     3421 tctaagaagg gcctgcagaa gatcctgaac gtgcagaaga agctgcctat catccagaag
     3481 atcatcatca tggactctaa gaccgactac cagggcttcc agagcatgta cacattcgtg
     3541 acatctcatc tgcctcctgg cttcaacgag tacgacttcg tgccagagtc tttcgacagg
     3601 gacaaaacca ttgccctgat catgaacagc tctgggtcta ccggcctgcc taagggcgtg
     3661 gccctgcctc atcgcaccgc ctgtgtgcgc ttctctcacg cccgcgaccc tattttcggc
     3721 aaccagatca tccccgacac cgctattctg agcgtggtgc cattccacca cggcttcggc
     3781 atgttcacca ccctgggcta cctgatttgc ggctttcggg tggtgctgat gtaccgcttc
     3841 gaggaggagc tgttcctgcg cagcctgcaa gactacaaaa ttcagtctgc cctgctggtg
     3901 ccaaccctgt tcagcttctt cgctaagagc accctgatcg acaagtacga cctgtctaac
     3961 ctgcacgaga ttgcctctgg cggcgcccca ctgtctaagg aggtgggcga agccgtggcc
     4021 aagcgctttc atctgccagg catccgccag ggctacggcc tgaccgagac aaccagcgcc
     4081 attctgatta ccccagaggg cgacgacaag cctggcgccg tgggcaaggt ggtgccattc
     4141 ttcgaggcca aggtggtgga cctggacacc ggcaagaccc tgggagtgaa ccagcgcggc
     4201 gagctgtgtg tgcgcggccc tatgattatg tccggctacg tgaataaccc tgaggccaca
     4261 aacgccctga tcgacaagga cggctggctg cactctggcg acattgccta ctgggacgag
     4321 gacgagcact tcttcatcgt ggaccgcctg aagtctctga tcaagtacaa gggctaccag
     4381 gtggccccag ccgagctgga gtctatcctg ctgcagcacc ctaacatttt cgacgccgga
     4441 gtggccggcc tgcccgacga cgatgccggc gagctgcctg ccgccgtcgt cgtgctggaa
     4501 cacggcaaga ccatgaccga gaaggagatc gtggactatg tggccagcca ggtgacaacc
     4561 gccaagaagc tgcgcggcgg agtggtgttc gtggacgagg tgcccaaggg cctgaccggc
     4621 aagctggacg cccgcaagat ccgcgagatc ctgatcaagg ctaagaaagg cggcaagatc
     4681 gccgtgtaat aattctagag tcggggcggc cggccgcttc gagcagacat gataagatac
     4741 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     4801 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     4861 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     4921 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc caggtggcac ttttcgggga
     4981 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5041 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5101 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5161 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5221 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5281 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5341 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5401 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5461 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5521 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5581 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     5641 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5701 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5761 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5821 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5881 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5941 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6001 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6061 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6121 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6181 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6241 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6301 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6361 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6421 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6481 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6541 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6601 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6661 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6721 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat ggctcgac
//
