LOCUS       sequence-243538-        7048 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  splicing reporter (with intron) with elements to shorten the 
            half-life of the luciferase protein as well as the luciferase mRNA..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7048)
  AUTHORS   Younis I, Berg M, Kaida D, Dittmar K, Wang C, Dreyfuss G
  TITLE     Rapid-response splicing reporter screens identify differential 
            regulators of constitutive and alternative splicing.
  JOURNAL   Mol Cell Biol. 2010 Apr;30(7):1718-28. doi: 10.1128/MCB.01301-09. 
            Epub 2010 Feb 1.
  PUBMED    20123975
REFERENCE   2  (bases 1 to 7048)
  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: 
            "10.1128/MCB.01301-09"; journalName: "Mol Cell Biol"; date: 
            "2010-04"; volume: "30"; issue: "7"; pages: "1718-28"
COMMENT     Sequence Label: CMV-LUC2CP/intron/ARE
FEATURES             Location/Qualifiers
     source          1..7048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(78..97)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        269..648
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        649..852
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     803..823
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     897..916
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        897..915
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      965..974
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          2315..2447
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2430..2449
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             2760..2807
                     /codon_start=1
                     /product="non-ORF yeast peptide conferring 
                     ubiquitin-dependent degradation"
                     /label=hCL1
                     /note="human codon-optimized"
                     /translation="ACKNWFSSLSHFVIHL"
     CDS             2811..2930
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    3003..3124
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3014..3033)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3068..3087
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3318..3675
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3318..3509
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3341..3361)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3526..3661
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3588..3607
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3706..4743
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4722..4730
                     /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    4767..4815
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4971..4988)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5142..5730)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5222..5241)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5930..6721,6722..6790))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    6895..6943
                     /note="synthetic polyadenylation signal"
     misc_feature    6957..7048
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6997..7016
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gcctcgagga tatcaagatc tcccgatccc
       61 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
      121 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
      181 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
      241 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
      301 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      361 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      421 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      481 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      541 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      601 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      661 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      721 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      781 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      841 ataagcagag ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat
      901 acgactcact atagggagac ccaagctggc tagttaagct tggcaatccg gtactgttgg
      961 taaagccacc atggaagatg ccaaaaacat taagaagggc ccagcgccat tctacccact
     1021 cgaagacggg accgccggcg agcagctgca caaagccatg aagcgctacg ccctggtgcc
     1081 cggcaccatc gcctttaccg acgcacatat cgaggtggac attacctacg ccgagtactt
     1141 cgagatgagc gttcggctgg cagaagctat gaagcgctat gggctgaata caaaccatcg
     1201 gatcgtggtg tgcagcgaga atagcttgca gttcttcatg cccgtgttgg gtgccctgtt
     1261 catcggtgtg gctgtggccc cagctaacga catctacaac gagcgcgagc tgctgaacag
     1321 catgggcatc agccagccca ccgtcgtatt cgtgagcaag aaagggctgc aaaagatcct
     1381 caacgtgcaa aagaagctac cgatcataca aaagatcatc atcatggata gcaagaccga
     1441 ctaccagggc ttccaaagca tgtacacctt cgtgacttcc catttgccac ccggcttcaa
     1501 cgagtacgac ttcgtgcccg agagcttcga ccgggacaaa accatcgccc tgatcatgaa
     1561 cagtagtggc agtaccggat tgcccaaggg cgtagcccta ccgcaccgca ccgcttgtgt
     1621 ccgattcagt catgcccgcg accccatctt cggcaaccag atcatccccg acaccgctat
     1681 cctcagcgtg gtgccatttc accacggctt cggcatgttc accacgctgg gctacttgat
     1741 ctgcggcttt cgggtcgtgc tcatgtaccg cttcgaggag gagctattct tgcgcagctt
     1801 gcaagactat aagattcaat ctgccctgct ggtgcccaca ctatttagct tcttcgctaa
     1861 gagcactctc atcgacaagt acgacctaag caacttgcac gagatcgcca gcggcggggc
     1921 gccgctcagc aaggaggtag gtgaggccgt ggccaaacgc ttccacctac caggcatccg
     1981 ccagggctac ggcctgacag aaacaaccag cgccattctg atcacccccg aaggggacga
     2041 caagcctggc gcagtaggca aggtggtgcc cttcttcgag gctaaggtgg tggacttgga
     2101 caccggtaag acactgggtg tgaaccagcg cggcgagctg tgcgtccgtg gccccatgat
     2161 catgagcggc tacgttaaca accccgaggc tacaaacgct ctcatcgaca aggacggctg
     2221 gctgcacagc ggcgacatcg cctactggga cgaggacgag cacttcttca tcgtggaccg
     2281 gctgaagagc ctgatcaaat acaagggcta ccaggtaagt atcaaggtta caagacaggt
     2341 ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata
     2401 ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggta gccccagccg
     2461 aactggagag catcctgctg caacacccca acatcttcga cgccggggtc gccggcctgc
     2521 ccgacgacga tgccggcgag ctgcccgccg cagtcgtcgt gctggaacac ggtaaaacca
     2581 tgaccgagaa ggagatcgtg gactatgtgg ccagccaggt tacaaccgcc aagaagctgc
     2641 gcggtggtgt tgtgttcgtg gacgaggtgc ctaaaggact gaccggcaag ttggacgccc
     2701 gcaagatccg cgagattctc attaaggcca agaagggcgg caagatcgcc gtgaattctg
     2761 cttgcaagaa ctggttcagt agcttaagcc actttgtgat ccaccttaac agccacggct
     2821 tccctcccga ggtggaggag caggccgccg gcaccctgcc catgagctgc gcccaggaga
     2881 gcggcatgga tagacaccct gctgcttgcg ccagcgccag gatcaacgtc taactgcagt
     2941 ctagatttat ttatttattt atttctagag tcggggcggc cggccgcttc gagcagacat
     3001 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     3061 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     3121 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     3181 tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtttgcgta
     3241 ttgggcgctc ttccgctgat ctgcgcagca ccatggcctg aaataacctc tgaaagagga
     3301 acttggttag ctaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     3361 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3421 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3481 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3541 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3601 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3661 ctaggctttt gcaaaaagct cgattcttct gacactagcg ccaccatgaa gaagcccgaa
     3721 ctcaccgcta ccagcgttga aaaatttctc atcgagaagt tcgacagtgt gagcgacctg
     3781 atgcagttgt cggagggcga agagagccga gccttcagct tcgatgtcgg cggacgcggc
     3841 tatgtactgc gggtgaatag ctgcgctgat ggcttctaca aagaccgcta cgtgtaccgc
     3901 cacttcgcca gcgctgcact acccatcccc gaagtgttgg acatcggcga gttcagcgag
     3961 agcctgacat actgcatcag tagacgcgcc caaggcgtta ctctccaaga cctccccgaa
     4021 acagagctgc ctgctgtgtt acagcctgtc gccgaagcta tggatgctat tgccgccgcc
     4081 gacctcagtc aaaccagcgg cttcggccca ttcgggcccc aaggcatcgg ccagtacaca
     4141 acctggcggg atttcatttg cgccattgct gatccccatg tctaccactg gcagaccgtg
     4201 atggacgaca ccgtgtccgc cagcgtagct caagccctgg acgaactgat gctgtgggcc
     4261 gaagactgtc ccgaggtgcg ccacctcgtc catgccgact tcggcagcaa caacgtcctg
     4321 accgacaacg gccgcatcac cgccgtaatc gactggtccg aagctatgtt cggggacagt
     4381 cagtacgagg tggccaacat cttcttctgg cggccctggc tggcttgcat ggagcagcag
     4441 actcgctact tcgagcgccg gcatcccgag ctggccggca gccctcgtct gcgagcctac
     4501 atgctgcgca tcggcctgga tcagctctac cagagcctcg tggacggcaa cttcgacgat
     4561 gctgcctggg ctcaaggccg ctgcgatgcc atcgtccgca gcggggccgg caccgtcggt
     4621 cgcacacaaa tcgctcgccg gagcgcagcc gtatggaccg acggctgcgt cgaggtgctg
     4681 gccgacagcg gcaaccgccg gcccagtaca cgaccgcgcg ctaaggaggt aggtcgagtt
     4741 taaactctag aaccggtcat ggccgcaata aaatatcttt attttcatta catctgtgtg
     4801 ttggtttttt gtgtgttcga actagatgct gtcgaccgat gcccttgaga gccttcaacc
     4861 cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct
     4921 tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact
     4981 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5041 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5101 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5161 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5221 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5281 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5341 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5401 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5461 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5521 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5581 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5641 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5701 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5761 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5821 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5881 gagtaaactt ggtctgacag cggccgcaaa tgctaaacca ctgcagtggt taccagtgct
     5941 tgatcagtga ggcaccgatc tcagcgatct gcctatttcg ttcgtccata gtggcctgac
     6001 tccccgtcgt gtagatcact acgattcgtg agggcttacc atcaggcccc agcgcagcaa
     6061 tgatgccgcg agagccgcgt tcaccggccc ccgatttgtc agcaatgaac cagccagcag
     6121 ggagggccga gcgaagaagt ggtcctgcta ctttgtccgc ctccatccag tctatgagct
     6181 gctgtcgtga tgctagagta agaagttcgc cagtgagtag tttccgaaga gttgtggcca
     6241 ttgctactgg catcgtggta tcacgctcgt cgttcggtat ggcttcgttc aactctggtt
     6301 cccagcggtc aagccgggtc acatgatcac ccatattatg aagaaatgca gtcagctcct
     6361 tagggcctcc gatcgttgtc agaagtaagt tggccgcggt gttgtcgctc atggtaatgg
     6421 cagcactaca caattctctt accgtcatgc catccgtaag atgcttttcc gtgaccggcg
     6481 agtactcaac caagtcgttt tgtgagtagt gtatacggcg accaagctgc tcttgcccgg
     6541 cgtctatacg ggacaacacc gcgccacata gcagtacttt gaaagtgctc atcatcggga
     6601 atcgttcttc ggggcggaaa gactcaagga tcttgccgct attgagatcc agttcgatat
     6661 agcccactct tgcacccagt tgatcttcag catcttttac tttcaccagc gtttcggggt
     6721 gtgcaaaaac aggcaagcaa aatgccgcaa agaagggaat gagtgcgaca cgaaaatgtt
     6781 ggatgctcat actcgtcctt tttcaatatt attgaagcat ttatcagggt tactagtacg
     6841 tctctcaagg ataagtaagt aatattaagg tacgggaggt attggacagg ccgcaataaa
     6901 atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agtactaaca
     6961 tacgctctcc atcaaaacaa aacgaaacaa aacaaactag caaaataggc tgtccccagt
     7021 gcaagtgcag gtgccagaac atttctct
//
