LOCUS       sequence-432478-        7041 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of Mbnl2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7041)
  AUTHORS   Weyn-Vanhentenryck SM, Feng H, Ustianenko D, Duffie R, Yan Q, Jacko 
            M, Martinez JC, Goodwin M, Zhang X, Hengst U, Lomvardas S, Swanson 
            MS, Zhang C
  TITLE     Precise temporal regulation of alternative splicing during neural 
            development.
  JOURNAL   Nat Commun. 2018 Jun 6;9(1):2189. doi: 10.1038/s41467-018-04559-0.
  PUBMED    29875359
REFERENCE   2  (bases 1 to 7041)
  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: "Nat 
            Commun."; date: "2018-06-6"; volume: "9(1):2189. doi"; pages: " 
            10.1038/s41467-018-04559-0"
COMMENT     Sequence Label: pCAGGS 3xFLAG Mbnl2
FEATURES             Location/Qualifiers
     source          1..7041
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     181..200
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    199..254
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     300..319
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     primer_bind     421..437
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     456..476
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        458..476
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    572..683
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     672..689
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     misc_feature    716..1304
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             822..833
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1231..1251
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(1311..1330)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          1313..1445
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     CDS             complement(2536..2556)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             complement(2563..2634)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     primer_bind     complement(2677..2696)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          2704..3720
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(2758..2780)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3721..3996)
                     /label=chicken beta-actin promoter
     enhancer        3998..4377
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4408..4512
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4513..4581,4582..5373)
                     /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(4731..4750)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5544..6132
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6033..6052
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6286..6303
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6371..6505
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6395..6414)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6449..6468
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6511..6706)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6525..6660
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    6765..6786
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6801..6806,6807..6824,6825..6831)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6839..6855
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6863..6879
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aaaaaaaatc taacctcaag tcaaggcttt tctatggaat aaggaatgga cagcaggggg
       61 ctgtttcata tactgatgac ctctttatag ccaacctttg ttcatggcag ccagcatatg
      121 ggcatatgtt gccaaactct aaaccaaata ctcattctga tgttttaaat gatttgccct
      181 cccatatgtc cttccgagtg agagacacaa aaaattccaa cacactattg caatgaaaat
      241 aaatttcctt tattagccag aagtcagatg ctcaaggggc ttcatgatgt ccccataatt
      301 tttggcagag ggaaaaagat ctcagtggta tttgtgagcc agggcattgg ccacaccagc
      361 caccaccttc tgataggcag cctgcacctg aggagtgaat tagcggataa caatttcaca
      421 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac
      481 aaaagctggg ctattgtctt cccaatcctc ccccttgctg tcctgcccca ccccaccccc
      541 cagaatagaa tgacacctac tcagacaatg cgatgcaatt tcctcatttt attaggaaag
      601 gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga
      661 tggctggcaa ctagaaggca cagtcgaggc tgatcgtacc gagctcgaat tccaggcggg
      721 gaggcggccc aaagggagat ccgactcgtc tgagggcgaa ggcgaagacg cggaagaggc
      781 cgcagagccg gcagcaggcc gcgggaagga aggtccgctg gattgagggc cgaagggacg
      841 tagcagaagg acgtcccgcg cagaatccag gtggcaacac aggcgagcag ccatggaaag
      901 gacgtcagct tccccgacaa caccacggaa ttgtcagtgc ccaacagccg agcccctgtc
      961 cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg caatagggag ggggaaagcg
     1021 aaagtcccgg aaaggagctg acaggtggtg gcaatgcccc aaccagtggg ggttgcgtca
     1081 gcaaacacag tgcacaccac gccacgttgc ctgacaacgg gccacaactc ctcataaaga
     1141 gacagcaacc aggatttata caaggaggag aaaatgaaag ccatacggga agcaatagca
     1201 tgatacaaag gcattaaagc agcgtatcca catagcgtaa aaggagcaac atagttaaga
     1261 ataccagtca atctttcaca aattttgtaa tccagaggtt gattgtcgac acctgtggag
     1321 agaaaggcaa agtggatgtc agtaagacca ataggtgcct atcagaaacg caagagtctt
     1381 ctctgtctcg acaagcccag tttctattgg tctccttaaa cctgtcttgt aaccttgata
     1441 cttacctgcg gcctcgagtt acttaagttt cagaattatc tgattggctg tggctgttgc
     1501 tgcgaagggg acacttgttg caggagttgt tgctgcagag acagtggcgg acgtagcgct
     1561 gtgcatcatg ggtactgtgg ggatgaacgc cgtgtgctgc tgcagctgcg cactggtcag
     1621 agcctgctgg tagtgcaaga cgctggggtt gaagaccgtg ctggccccgt tgcttttttc
     1681 aagtgcttgt ctctttggta agggatgaag agcacccgga gggaaggcca tgactgtggc
     1741 cgcggccgcg gctgcctggg cggccaccgc ggcctggttg gcttggtgct gcgcagcttt
     1801 gattttggcc tgcaagtgtg caggagggtg aaaatatttg catttctccc tcatgcaacg
     1861 cccctttatg taatccatac aaacggttac ggtgttgtcg tttgtgtcga tcatggtgct
     1921 gtctgccggg tgtgcaaagc ggcagtctgt ctctccccgg gcacagtttc ctcgctggaa
     1981 ctccctgcat acctccagtt tatcagtcct gagaagtttc tgagttgcag ttgagcccgg
     2041 gacagtgacc ggtggacttc cgggaacaat gaccggtgta gtgggtagaa cctctgttgg
     2101 gactaaccca actccagggg tgacaggcgc taagtaagga gcaaagctaa tagccgcatt
     2161 tgtccctatg gtgggaccta caggaaaagt gggcacagga tggagcggcg ttcctggaaa
     2221 cataaattgc atctgctggg caagcatcgc tgctgcagtt ttttgctgga tcaaattgtt
     2281 cctcccatta atctctagct gggtttttaa gtgtgtcgga ggatgaagat atttgcagtt
     2341 ctctcttgaa cagcggccct tgagggaatc aaagcaggca attactcttc cattttcaac
     2401 ctggcaactt ttgggggggt gagcaaactt gcattcttcg tcggagcgtg agcacgttcc
     2461 tctctggtac tgtctgcaga cctccagcgt cagccacttt gtgtctctca cgggggcaac
     2521 gttcaaggca ccggtcgatt ggaagtagag gttctcgcta gctttatcgt catcgtcttt
     2581 gtagtccttg tcgtcgtcgt ccttgtaatc cttatcatca tcatctttat agtcgctagc
     2641 catggtaccg aattctttgc caaaatgatg agacagcaca ataaccagca cgttgcccag
     2701 gagctgtagg aaaaagaaga aggcatgaac atggttagca gaggctctag agccgccggt
     2761 cacacgccag aagccgaacc ccgccctgcc ccgtcccccc cgaaggcagc cgtccccccg
     2821 cggacagccc cgaggctgga gagggagaag gggacggcgg cgcggcgacg cacgaaggcc
     2881 ctccccgccc atttccttcc tgccggcgcc gcaccgcttc gccccgcgcc cgctagaggg
     2941 ggtgcggcgg cgcctcccag atttcggctc cgcacagatt tgggacaaag gaagtccctg
     3001 cgccctctcg cacgattacc ataaaaggca atggctgcgg ctcgccgcgc ctcgacagcc
     3061 gccggcgctc cgggggccgc cgcgcccctc ccccgagccc tccccggccc gaggcggccc
     3121 cgccccgccc ggcaccccca cctgccgcca ccccccgccc ggcacggcga gccccgcgcc
     3181 acgccccgta cggagccccg cacccgaagc cgggccgtgc tcagcaactc ggggaggggg
     3241 gtgcaggggg ggttgcagcc cgaccgacgc gcccacaccc cctgctcacc cccccacgca
     3301 cacaccccgc acgcagcctt tgttcccctc gcagcccccc ccgcaccgcg gggcaccgcc
     3361 cccggccgcg ctcccctcgc gcacactgcg gagcgcacaa agccccgcgc cgcgcccgca
     3421 gcgctcacag ccgccgggca gcgcggagcc gcacgcggcg ctccccacgc acacacacac
     3481 gcacgcaccc cccgagccgc tccccccgca caaagggccc tcccggagcc cctcaaggct
     3541 ttcacgcagc cacagaaaag aaacaagccg tcattaaacc aagcgctaat tacagcccgg
     3601 aggagaaggg ccgtcccgcc cgctcacctg tgggagtaac gcggtcagtc agagccgggg
     3661 cgggcggcgc gaggcggcgg cggagcgggg cacggggcga aggcagcgcg cagcgactcc
     3721 cgcccgccgc gcgcttcgct ttttataggg ccgccgccgc cgccgcctcg ccataaaagg
     3781 aaactttcgg agcgcgccgc tctgattggc tgccgccgca cctctccgcc tcgccccgcc
     3841 ccgcccctcg ccccgccccg ccccgcctgg cgcgcscccc cccccccccc gcccccatcg
     3901 ctgcacaaaa taattaaaaa ataaataaat acaaaattgg gggtggggag gggggggaga
     3961 tggggagagt gaagcagaac gtggggctca cctcgaccat ggtaatagcg atgactaata
     4021 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     4081 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     4141 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     4201 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     4261 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     4321 tatgaactaa tgaccccgta attgattact attaataact agtcaataat caatgtcgac
     4381 ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4441 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4501 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4561 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4621 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4681 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4741 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4801 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     4861 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     4921 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     4981 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5041 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5101 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5161 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5221 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5281 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5341 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5401 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5461 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5521 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5581 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5641 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5701 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5761 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5821 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5881 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5941 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6001 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6061 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6121 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6181 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6241 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     6301 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     6361 taatgcagcg gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     6421 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     6481 cattataagc tgcaataaac aagttagctt tttgcaaaag cctaggcctc caaaaaagcc
     6541 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     6601 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     6661 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcggat ccgctggcac
     6721 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     6781 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     6841 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttggg
     6901 ctgcaggtcg agggatctcc ataagagaag agggacagct atgactggga gtagtcagga
     6961 gaggaggaaa aatctggcta gtaaaacatg taaggaaaat tttagggatg ttaaagaaaa
     7021 aaataacaca aaacaaaata t
//
