LOCUS       sequence-331323-        7049 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector, Luciferase reporter to test activity of
            miRNA (miR-137).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7049)
  TITLE     Karreth lab plasmids
REFERENCE   2  (bases 1 to 7049)
  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"
COMMENT     Sequence Label: psiCHECK2-miR-137
FEATURES             Location/Qualifiers
     source          1..7049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..413
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        56..247
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(79..99)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      264..399
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          483..615
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     598..617
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        660..678
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      682..691
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             688..1623
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1681..1729
                     /note="synthetic polyadenylation signal"
     promoter        1738..2489
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      2519..2528
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2525..4177
                     /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             3484..3492
                     /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    4221..4342
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4232..4251)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4286..4305
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     mobile_element  4531..5298
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     CDS             join(5357..5425,5426..6217)
                     /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(5575..5594)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6388..6976
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6877..6896
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag
       61 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
      121 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
      181 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
      241 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
      301 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
      361 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagcttg
      421 attcttctga cacaacagtc tcgaacttaa gctgcagaag ttggtcgtga ggcactgggc
      481 aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga
      541 gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg
      601 cctttctctc cacaggtgtc cactcccagt tcaattacag ctcttaaggc tagagtactt
      661 aatacgactc actataggct agccaccatg gcttccaagg tgtacgaccc cgagcaacgc
      721 aaacgcatga tcactgggcc tcagtggtgg gctcgctgca agcaaatgaa cgtgctggac
      781 tccttcatca actactatga ttccgagaag cacgccgaga acgccgtgat ttttctgcat
      841 ggtaacgctg cctccagcta cctgtggagg cacgtcgtgc ctcacatcga gcccgtggct
      901 agatgcatca tccctgatct gatcggaatg ggtaagtccg gcaagagcgg gaatggctca
      961 tatcgcctcc tggatcacta caagtacctc accgcttggt tcgagctgct gaaccttcca
     1021 aagaaaatca tctttgtggg ccacgactgg ggggcttgtc tggcctttca ctactcctac
     1081 gagcaccaag acaagatcaa ggccatcgtc catgctgaga gtgtcgtgga cgtgatcgag
     1141 tcctgggacg agtggcctga catcgaggag gatatcgccc tgatcaagag cgaagagggc
     1201 gagaaaatgg tgcttgagaa taacttcttc gtcgagacca tgctcccaag caagatcatg
     1261 cggaaactgg agcctgagga gttcgctgcc tacctggagc cattcaagga gaagggcgag
     1321 gttagacggc ctaccctctc ctggcctcgc gagatccctc tcgttaaggg aggcaagccc
     1381 gacgtcgtcc agattgtccg caactacaac gcctaccttc gggccagcga cgatctgcct
     1441 aagatgttca tcgagtccga ccctgggttc ttttccaacg ctattgtcga gggagctaag
     1501 aagttcccta acaccgagtt cgtgaaggtg aagggcctcc acttcagcca ggaggacgct
     1561 ccagatgaaa tgggtaagta catcaagagc ttcgtggagc gcgtgctgaa gaacgagcag
     1621 taattctagg cgatcgctcg acctacgcgt attcttaagc aataagcggc cgctggccgc
     1681 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga ggatctaaat
     1741 gagtcttcgg acctcgcggg ggccgcttaa gcggtggtta gggtttgtct gacgcggggg
     1801 gagggggaag gaacgaaaca ctctcattcg gaggcggctc ggggtttggt cttggtggcc
     1861 acgggcacgc agaagagcgc cgcgatcctc ttaagcaccc ccccgccctc cgtggaggcg
     1921 ggggtttggt cggcgggtgg taactggcgg gccgctgact cgggcgggtc gcgcgcccca
     1981 gagtgtgacc ttttcggtct gctcgcagac ccccgggcgg cgccgccgcg gcggcgacgg
     2041 gctcgctggg tcctaggctc catggggacc gtatacgtgg acaggctctg gagcatccgc
     2101 acgactgcgg tgatattacc ggagaccttc tgcgggacga gccgggtcac gcggctgacg
     2161 cggagcgtcc gttgggcgac aaacaccagg acggggcaca ggtacactat cttgtcaccc
     2221 ggaggcgcga gggactgcag gagcttcagg gagtggcgca gctgcttcat ccccgtggcc
     2281 cgttgctcgc gtttgctggc ggtgtccccg gaagaaatat atttgcatgt ctttagttct
     2341 atgatgacac aaaccccgcc cagcgtcttg tcattggcga attcgaacac gcagatgcag
     2401 tcggggcggc gcggtcccag gtccacttcg catattaagg tgacgcgtgt ggcctcgaac
     2461 accgagcgac cctgcagcga cccgcttaaa agcttggcat tccggtactg ttggtaaagc
     2521 caccatggcc gatgctaaga acattaagaa gggccctgct cccttctacc ctctggagga
     2581 tggcaccgct ggcgagcagc tgcacaaggc catgaagagg tatgccctgg tgcctggcac
     2641 cattgccttc accgatgccc acattgaggt ggacatcacc tatgccgagt acttcgagat
     2701 gtctgtgcgc ctggccgagg ccatgaagag gtacggcctg aacaccaacc accgcatcgt
     2761 ggtgtgctct gagaactctc tgcagttctt catgccagtg ctgggcgccc tgttcatcgg
     2821 agtggccgtg gcccctgcta acgacattta caacgagcgc gagctgctga acagcatggg
     2881 catttctcag cctaccgtgg tgttcgtgtc taagaagggc ctgcagaaga tcctgaacgt
     2941 gcagaagaag ctgcctatca tccagaagat catcatcatg gactctaaga ccgactacca
     3001 gggcttccag agcatgtaca cattcgtgac atctcatctg cctcctggct tcaacgagta
     3061 cgacttcgtg ccagagtctt tcgacaggga caaaaccatt gccctgatca tgaacagctc
     3121 tgggtctacc ggcctgccta agggcgtggc cctgcctcat cgcaccgcct gtgtgcgctt
     3181 ctctcacgcc cgcgacccta ttttcggcaa ccagatcatc cccgacaccg ctattctgag
     3241 cgtggtgcca ttccaccacg gcttcggcat gttcaccacc ctgggctacc tgatttgcgg
     3301 ctttcgggtg gtgctgatgt accgcttcga ggaggagctg ttcctgcgca gcctgcaaga
     3361 ctacaaaatt cagtctgccc tgctggtgcc aaccctgttc agcttcttcg ctaagagcac
     3421 cctgatcgac aagtacgacc tgtctaacct gcacgagatt gcctctggcg gcgccccact
     3481 gtctaaggag gtgggcgaag ccgtggccaa gcgctttcat ctgccaggca tccgccaggg
     3541 ctacggcctg accgagacaa ccagcgccat tctgattacc ccagagggcg acgacaagcc
     3601 tggcgccgtg ggcaaggtgg tgccattctt cgaggccaag gtggtggacc tggacaccgg
     3661 caagaccctg ggagtgaacc agcgcggcga gctgtgtgtg cgcggcccta tgattatgtc
     3721 cggctacgtg aataaccctg aggccacaaa cgccctgatc gacaaggacg gctggctgca
     3781 ctctggcgac attgcctact gggacgagga cgagcacttc ttcatcgtgg accgcctgaa
     3841 gtctctgatc aagtacaagg gctaccaggt ggccccagcc gagctggagt ctatcctgct
     3901 gcagcaccct aacattttcg acgccggagt ggccggcctg cccgacgacg atgccggcga
     3961 gctgcctgcc gccgtcgtcg tgctggaaca cggcaagacc atgaccgaga aggagatcgt
     4021 ggactatgtg gccagccagg tgacaaccgc caagaagctg cgcggcggag tggtgttcgt
     4081 ggacgaggtg cccaagggcc tgaccggcaa gctggacgcc cgcaagatcc gcgagatcct
     4141 gatcaaggct aagaaaggcg gcaagatcgc cgtgtaataa ttctagagtc ggggcggccg
     4201 gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     4261 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     4321 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     4381 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga
     4441 taaggatcca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4501 ctaaatacat tcaaatatgt atccgctcat ggtgatgctg ccaacttact gatttagtgt
     4561 atgatggtgt ttttgaggtg ctccagtggc ttctgtttct atcagctgtc cctcctgttc
     4621 agctactgac ggggtggtgc gtaacggcaa aagcaccgcc ggacatcagc gctatctctg
     4681 ctctcactgc cgtaaaacat ggcaactgca gttcacttac accgcttctc aacccggtac
     4741 gcaccagaaa atcattgata tggccatgaa tggcgttgga tgccgggcaa cagcccgcat
     4801 tatgggcgtt ggcctcaaca cgattttacg tcacttaaaa aactcaggcc gcagtcggta
     4861 acctcgcgca tacagccggg cagtgacgtc atcgtctgcg cggaaatgga cgaacagtgg
     4921 ggctatgtcg gggctaaatc gcgccagcgc tggctgtttt acgcgtatga cagtctccgg
     4981 aagacggttg ttgcgcacgt attcggtgaa cgcactatgg cgacgctggg gcgtcttatg
     5041 agcctgctgt caccctttga cgtggtgata tggatgacgg atggctggcc gctgtatgaa
     5101 tcccgcctga agggaaagct gcacgtaatc agcaagcgat atacgcagcg aattgagcgg
     5161 cataacctga atctgaggca gcacctggca cggctgggac ggaagtcgct gtcgttctca
     5221 aaatcggtgg agctgcatga caaagtcatc gggcattatc tgaacataaa acactatcaa
     5281 taagttggag tcattacctc cgctcatgag acaataaccc tgataaatgc ttcaataata
     5341 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     5401 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5461 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5521 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5581 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5641 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5701 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5761 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5821 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5881 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5941 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     6001 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     6061 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     6121 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     6181 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     6241 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6301 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6361 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6421 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6481 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6541 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6601 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6661 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6721 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6781 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6841 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6901 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6961 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     7021 gccttttgct cacatggctc gacagatct
//
