LOCUS       sequence-99091-e        7231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  splicing reporter construct containing MAG mini-gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7231)
  AUTHORS   Zearfoss NR, Johnson ES, Ryder SP
  TITLE     hnRNP A1 and secondary structure coordinate alternative splicing of 
            Mag.
  JOURNAL   RNA. 2013 Jul;19(7):948-57. doi: 10.1261/rna.036780.112. Epub 2013 
            May 23.
  PUBMED    23704325
REFERENCE   2  (bases 1 to 7231)
  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"; doi: 
            "10.1261/rna.036780.112"; journalName: "RNA"; date: "2013-07"; 
            volume: "19"; issue: "7"; pages: "948-57"
COMMENT     Sequence Label: psc2_md1
FEATURES             Location/Qualifiers
     source          1..7231
                     /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    1688..1736
                     /note="synthetic polyadenylation signal"
     promoter        1745..2496
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             3493..5142
                     /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="ADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAH
                     IEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPA
                     NDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSM
                     YTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHAR
                     DPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKI
                     QSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYG
                     LTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSG
                     YVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILL
                     QHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVF
                     VDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     RBS             4449..4457
                     /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    5186..5307
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5197..5216)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5251..5270
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5440..5544
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5545..5613,5614..6405)
                     /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(5763..5782)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6576..7164
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7065..7084
                     /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 ggcttgattc 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 cgctcgagcc cgggaattcg tttaaaccta gagcggccgc
     1681 tggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgagga
     1741 tctaaatgag tcttcggacc tcgcgggggc cgcttaagcg gtggttaggg tttgtctgac
     1801 gcggggggag ggggaaggaa cgaaacactc tcattcggag gcggctcggg gtttggtctt
     1861 ggtggccacg ggcacgcaga agagcgccgc gatcctctta agcacccccc cgccctccgt
     1921 ggaggcgggg gtttggtcgg cgggtggtaa ctggcgggcc gctgactcgg gcgggtcgcg
     1981 cgccccagag tgtgaccttt tcggtctgct cgcagacccc cgggcggcgc cgccgcggcg
     2041 gcgacgggct cgctgggtcc taggctccat ggggaccgta tacgtggaca ggctctggag
     2101 catccgcacg actgcggtga tattaccgga gaccttctgc gggacgagcc gggtcacgcg
     2161 gctgacgcgg agcgtccgtt gggcgacaaa caccaggacg gggcacaggt acactatctt
     2221 gtcacccgga ggcgcgaggg actgcaggag cttcagggag tggcgcagct gcttcatccc
     2281 cgtggcccgt tgctcgcgtt tgctggcggt gtccccggaa gaaatatatt tgcatgtctt
     2341 tagttctatg atgacacaaa ccccgcccag cgtcttgtca ttggcgaatt cgaacacgca
     2401 gatgcagtcg gggcggcgcg gtcccaggtc cacttcgcat attaaggtga cgcgtgtggc
     2461 ctcgaacacc gagcgaccct gcagcgaccc gcttaaaagc ttgggatgga ggtgagaggg
     2521 ccacacccct gtggctccca cttagcttgg gaaggacagg atggcaaggt tgctaagggt
     2581 gggggtggga caaaagggag gccagactca tgggtgtttg tagggctggg ctggggctag
     2641 agtttgggga ggaggctcct tgctgaagtt atgcaggagg gttgagggaa tggggcctgg
     2701 gtagccctgc cctccctgcc tgtgcctgtg actgcgtttc cttcttcaat agtccagaga
     2761 ggtctctacc cgggattgtc actgagagcc ccaggaggta ggtcccgggg gcctcagtgt
     2821 gccagcctcc gggccctagc tgggcttcgg ggttggtgtg catgtttgtg tgtgttccag
     2881 tcatgtgtcc tcagcactct ttggttaatc ccagccttgc tcagcagccc tggccttgtc
     2941 ttttctgttc cttcgctgat cccctttttc tgtctgtggc cactgcccta ttgtgtgtcc
     3001 aaatgctagt ggggtggggg tcaggagtag agagctggag aagtggggaa tcacaccacc
     3061 aggatcccct caactgagag aagccctatc atatccaaag agagaacggt ttactgaatg
     3121 gggctctttg caagcttcca aaagggtggt ccaaccaaga tttaatctag agaggtgttc
     3181 tgggtccagt catgcaccct taaagttatt ttctcaaaga gtgcttcttg gaacctggga
     3241 ttcctagggc aaacatcaag gtgagcagtg agatagtggt tttactggca tcagcacttg
     3301 caaagcccct ccccccaccc ccaagtagga catgatgact gtagaactcc ctgagctggt
     3361 ggcccctgag aaaaaggtgc cagcagccca gcctttgcat gggaccgtca gagagagcca
     3421 cgctgacggg ttgtcttctc ctctccatag gcgtctcaag cttggcattc cggtactgtt
     3481 ggtaaagcca ccgccgatgc taagaacatt aagaagggcc ctgctccctt ctaccctctg
     3541 gaggatggca ccgctggcga gcagctgcac aaggccatga agaggtatgc cctggtgcct
     3601 ggcaccattg ccttcaccga tgcccacatt gaggtggaca tcacctatgc cgagtacttc
     3661 gagatgtctg tgcgcctggc cgaggccatg aagaggtacg gcctgaacac caaccaccgc
     3721 atcgtggtgt gctctgagaa ctctctgcag ttcttcatgc cagtgctggg cgccctgttc
     3781 atcggagtgg ccgtggcccc tgctaacgac atttacaacg agcgcgagct gctgaacagc
     3841 atgggcattt ctcagcctac cgtggtgttc gtgtctaaga agggcctgca gaagatcctg
     3901 aacgtgcaga agaagctgcc tatcatccag aagatcatca tcatggactc taagaccgac
     3961 taccagggct tccagagcat gtacacattc gtgacatctc atctgcctcc tggcttcaac
     4021 gagtacgact tcgtgccaga gtctttcgac agggacaaaa ccattgccct gatcatgaac
     4081 agctctgggt ctaccggcct gcctaagggc gtggccctgc ctcatcgcac cgcctgtgtg
     4141 cgcttctctc acgcccgcga ccctattttc ggcaaccaga tcatccccga caccgctatt
     4201 ctgagcgtgg tgccattcca ccacggcttc ggcatgttca ccaccctggg ctacctgatt
     4261 tgcggctttc gggtggtgct gatgtaccgc ttcgaggagg agctgttcct gcgcagcctg
     4321 caagactaca aaattcagtc tgccctgctg gtgccaaccc tgttcagctt cttcgctaag
     4381 agcaccctga tcgacaagta cgacctgtct aacctgcacg agattgcctc tggcggcgcc
     4441 ccactgtcta aggaggtggg cgaagccgtg gccaagcgct ttcatctgcc aggcatccgc
     4501 cagggctacg gcctgaccga gacaaccagc gccattctga ttaccccaga gggcgacgac
     4561 aagcctggcg ccgtgggcaa ggtggtgcca ttcttcgagg ccaaggtggt ggacctggac
     4621 accggcaaga ccctgggagt gaaccagcgc ggcgagctgt gtgtgcgcgg ccctatgatt
     4681 atgtccggct acgtgaataa ccctgaggcc acaaacgccc tgatcgacaa ggacggctgg
     4741 ctgcactctg gcgacattgc ctactgggac gaggacgagc acttcttcat cgtggaccgc
     4801 ctgaagtctc tgatcaagta caagggctac caggtggccc cagccgagct ggagtctatc
     4861 ctgctgcagc accctaacat tttcgacgcc ggagtggccg gcctgcccga cgacgatgcc
     4921 ggcgagctgc ctgccgccgt cgtcgtgctg gaacacggca agaccatgac cgagaaggag
     4981 atcgtggact atgtggccag ccaggtgaca accgccaaga agctgcgcgg cggagtggtg
     5041 ttcgtggacg aggtgcccaa gggcctgacc ggcaagctgg acgcccgcaa gatccgcgag
     5101 atcctgatca aggctaagaa aggcggcaag atcgccgtgt aataattcta gagtcggggc
     5161 ggccggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     5221 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     5281 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     5341 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa
     5401 atcgataagg atccaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5461 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5521 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5581 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5641 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5701 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5761 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5821 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5881 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5941 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     6001 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     6061 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     6121 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6181 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6241 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6301 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6361 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6421 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6481 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6541 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6601 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6661 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6721 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6781 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6841 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6901 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6961 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7021 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7081 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7141 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7201 ttttgctggc cttttgctca catggctcga c
//
