LOCUS       sequence-212900-        6915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Splicing reporter control (no 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 6915)
  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 6915)
  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/ARE
FEATURES             Location/Qualifiers
     source          1..6915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    46..94
                     /note="synthetic polyadenylation signal"
     misc_feature    108..199
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     148..167
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     primer_bind     complement(277..296)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        468..847
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        848..1051
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1002..1022
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1096..1115
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1096..1114
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1164..1173
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1170..2819
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             2826..2873
                     /codon_start=1
                     /product="non-ORF yeast peptide conferring 
                     ubiquitin-dependent degradation"
                     /label=hCL1
                     /note="human codon-optimized"
                     /translation="ACKNWFSSLSHFVIHL"
     CDS             2877..2996
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    3069..3190
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3080..3099)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3134..3153
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3384..3741
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3384..3575
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3407..3427)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3592..3727
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3654..3673
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3772..4809
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4788..4796
                     /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    4833..4881
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5037..5054)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5208..5796)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5288..5307)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5996..6787,6788..6856))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 gataagtaag taatattaag gtacgggagg tattggacag gccgcaataa aatatcttta
       61 ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagtactaac atacgctctc
      121 catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca
      181 ggtgccagaa catttctctg gcctaactgg ccggtacctg agctcgctag cctcgaggat
      241 atcaagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
      301 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
      361 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
      421 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt
      481 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      541 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      601 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      661 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      721 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      781 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      841 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      901 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      961 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     1021 tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac ccactgctta
     1081 ctggcttatc gaaattaata cgactcacta tagggagacc caagctggct agttaagctt
     1141 ggcaatccgg tactgttggt aaagccacca tggaagatgc caaaaacatt aagaagggcc
     1201 cagcgccatt ctacccactc gaagacggga ccgccggcga gcagctgcac aaagccatga
     1261 agcgctacgc cctggtgccc ggcaccatcg cctttaccga cgcacatatc gaggtggaca
     1321 ttacctacgc cgagtacttc gagatgagcg ttcggctggc agaagctatg aagcgctatg
     1381 ggctgaatac aaaccatcgg atcgtggtgt gcagcgagaa tagcttgcag ttcttcatgc
     1441 ccgtgttggg tgccctgttc atcggtgtgg ctgtggcccc agctaacgac atctacaacg
     1501 agcgcgagct gctgaacagc atgggcatca gccagcccac cgtcgtattc gtgagcaaga
     1561 aagggctgca aaagatcctc aacgtgcaaa agaagctacc gatcatacaa aagatcatca
     1621 tcatggatag caagaccgac taccagggct tccaaagcat gtacaccttc gtgacttccc
     1681 atttgccacc cggcttcaac gagtacgact tcgtgcccga gagcttcgac cgggacaaaa
     1741 ccatcgccct gatcatgaac agtagtggca gtaccggatt gcccaagggc gtagccctac
     1801 cgcaccgcac cgcttgtgtc cgattcagtc atgcccgcga ccccatcttc ggcaaccaga
     1861 tcatccccga caccgctatc ctcagcgtgg tgccatttca ccacggcttc ggcatgttca
     1921 ccacgctggg ctacttgatc tgcggctttc gggtcgtgct catgtaccgc ttcgaggagg
     1981 agctattctt gcgcagcttg caagactata agattcaatc tgccctgctg gtgcccacac
     2041 tatttagctt cttcgctaag agcactctca tcgacaagta cgacctaagc aacttgcacg
     2101 agatcgccag cggcggggcg ccgctcagca aggaggtagg tgaggccgtg gccaaacgct
     2161 tccacctacc aggcatccgc cagggctacg gcctgacaga aacaaccagc gccattctga
     2221 tcacccccga aggggacgac aagcctggcg cagtaggcaa ggtggtgccc ttcttcgagg
     2281 ctaaggtggt ggacttggac accggtaaga cactgggtgt gaaccagcgc ggcgagctgt
     2341 gcgtccgtgg ccccatgatc atgagcggct acgttaacaa ccccgaggct acaaacgctc
     2401 tcatcgacaa ggacggctgg ctgcacagcg gcgacatcgc ctactgggac gaggacgagc
     2461 acttcttcat cgtggaccgg ctgaagagcc tgatcaaata caagggctac caggtagccc
     2521 cagccgaact ggagagcatc ctgctgcaac accccaacat cttcgacgcc ggggtcgccg
     2581 gcctgcccga cgacgatgcc ggcgagctgc ccgccgcagt cgtcgtgctg gaacacggta
     2641 aaaccatgac cgagaaggag atcgtggact atgtggccag ccaggttaca accgccaaga
     2701 agctgcgcgg tggtgttgtg ttcgtggacg aggtgcctaa aggactgacc ggcaagttgg
     2761 acgcccgcaa gatccgcgag attctcatta aggccaagaa gggcggcaag atcgccgtga
     2821 attctgcttg caagaactgg ttcagtagct taagccactt tgtgatccac cttaacagcc
     2881 acggcttccc tcccgaggtg gaggagcagg ccgccggcac cctgcccatg agctgcgccc
     2941 aggagagcgg catggataga caccctgctg cttgcgccag cgccaggatc aacgtctaac
     3001 tgcagtctag atttatttat ttatttattt ctagagtcgg ggcggccggc cgcttcgagc
     3061 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     3121 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     3181 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     3241 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatccgtt
     3301 tgcgtattgg gcgctcttcc gctgatctgc gcagcaccat ggcctgaaat aacctctgaa
     3361 agaggaactt ggttagctac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3421 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3481 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3541 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3601 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3661 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3721 ggaggcctag gcttttgcaa aaagctcgat tcttctgaca ctagcgccac catgaagaag
     3781 cccgaactca ccgctaccag cgttgaaaaa tttctcatcg agaagttcga cagtgtgagc
     3841 gacctgatgc agttgtcgga gggcgaagag agccgagcct tcagcttcga tgtcggcgga
     3901 cgcggctatg tactgcgggt gaatagctgc gctgatggct tctacaaaga ccgctacgtg
     3961 taccgccact tcgccagcgc tgcactaccc atccccgaag tgttggacat cggcgagttc
     4021 agcgagagcc tgacatactg catcagtaga cgcgcccaag gcgttactct ccaagacctc
     4081 cccgaaacag agctgcctgc tgtgttacag cctgtcgccg aagctatgga tgctattgcc
     4141 gccgccgacc tcagtcaaac cagcggcttc ggcccattcg ggccccaagg catcggccag
     4201 tacacaacct ggcgggattt catttgcgcc attgctgatc cccatgtcta ccactggcag
     4261 accgtgatgg acgacaccgt gtccgccagc gtagctcaag ccctggacga actgatgctg
     4321 tgggccgaag actgtcccga ggtgcgccac ctcgtccatg ccgacttcgg cagcaacaac
     4381 gtcctgaccg acaacggccg catcaccgcc gtaatcgact ggtccgaagc tatgttcggg
     4441 gacagtcagt acgaggtggc caacatcttc ttctggcggc cctggctggc ttgcatggag
     4501 cagcagactc gctacttcga gcgccggcat cccgagctgg ccggcagccc tcgtctgcga
     4561 gcctacatgc tgcgcatcgg cctggatcag ctctaccaga gcctcgtgga cggcaacttc
     4621 gacgatgctg cctgggctca aggccgctgc gatgccatcg tccgcagcgg ggccggcacc
     4681 gtcggtcgca cacaaatcgc tcgccggagc gcagccgtat ggaccgacgg ctgcgtcgag
     4741 gtgctggccg acagcggcaa ccgccggccc agtacacgac cgcgcgctaa ggaggtaggt
     4801 cgagtttaaa ctctagaacc ggtcatggcc gcaataaaat atctttattt tcattacatc
     4861 tgtgtgttgg ttttttgtgt gttcgaacta gatgctgtcg accgatgccc ttgagagcct
     4921 tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
     4981 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc cgcttcctcg
     5041 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5101 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5161 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5221 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5281 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5341 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5401 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5461 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5521 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5581 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5641 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5701 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5761 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5821 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5881 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5941 atatatgagt aaacttggtc tgacagcggc cgcaaatgct aaaccactgc agtggttacc
     6001 agtgcttgat cagtgaggca ccgatctcag cgatctgcct atttcgttcg tccatagtgg
     6061 cctgactccc cgtcgtgtag atcactacga ttcgtgaggg cttaccatca ggccccagcg
     6121 cagcaatgat gccgcgagag ccgcgttcac cggcccccga tttgtcagca atgaaccagc
     6181 cagcagggag ggccgagcga agaagtggtc ctgctacttt gtccgcctcc atccagtcta
     6241 tgagctgctg tcgtgatgct agagtaagaa gttcgccagt gagtagtttc cgaagagttg
     6301 tggccattgc tactggcatc gtggtatcac gctcgtcgtt cggtatggct tcgttcaact
     6361 ctggttccca gcggtcaagc cgggtcacat gatcacccat attatgaaga aatgcagtca
     6421 gctccttagg gcctccgatc gttgtcagaa gtaagttggc cgcggtgttg tcgctcatgg
     6481 taatggcagc actacacaat tctcttaccg tcatgccatc cgtaagatgc ttttccgtga
     6541 ccggcgagta ctcaaccaag tcgttttgtg agtagtgtat acggcgacca agctgctctt
     6601 gcccggcgtc tatacgggac aacaccgcgc cacatagcag tactttgaaa gtgctcatca
     6661 tcgggaatcg ttcttcgggg cggaaagact caaggatctt gccgctattg agatccagtt
     6721 cgatatagcc cactcttgca cccagttgat cttcagcatc ttttactttc accagcgttt
     6781 cggggtgtgc aaaaacaggc aagcaaaatg ccgcaaagaa gggaatgagt gcgacacgaa
     6841 aatgttggat gctcatactc gtcctttttc aatattattg aagcatttat cagggttact
     6901 agtacgtctc tcaag
//
