LOCUS       sequence-192193-        6443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of Flag-tagged human MBNL3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6443)
  AUTHORS   Orengo JP, Bundman D, Cooper TA
  TITLE     A bichromatic fluorescent reporter for cell-based screens of 
            alternative splicing.
  JOURNAL   Nucleic Acids Res. 2006;34(22):e148. Epub 2006 Nov 16.
  PUBMED    17142220
REFERENCE   2  (bases 1 to 6443)
  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: "Nucleic 
            Acids Res."; date: "2006"; volume: "34(22)"; pages: "e148. Epub 2006
            Nov 16"
COMMENT     Sequence Label: FlagMBNL3
FEATURES             Location/Qualifiers
     source          1..6443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             922..945
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(1843..1854)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(2034..2051)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2040..2264
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2310..2738
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2397..2416)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2607..2628
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2747..2767)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2752..3081
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2932..3067
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2994..3013
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3148..3942
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3202..3221)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3812..3831
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4116..4237
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4153..4172)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4207..4226
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4286..4302)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4286..4302)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4299..4321)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4310..4326
                     /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)."
     promoter        complement(join(4334..4340,4341..4358,4359..4364))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4379..4400
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4517..4534)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4688..5276)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4768..4787)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5447..6238,6239..6307))
                     /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     6070..6089
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6308..6412)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttccat ggattacaag gatgacgatg acaaggggaa ttctatgacg
      961 gctgtcaatg ttgccctgat tcgtgatacc aagtggctga ctttagaagt ctgtagagaa
     1021 tttcagagag gaacttgctc tcgagctgat gcagattgca agtttgccca tccaccaaga
     1081 gtttgccatg tggaaaatgg tcgtgtggtg gcctgttttg attctctaaa gggtcggtgt
     1141 acccgagaga actgcaagta ccttcaccct cctccacact taaaaacgca gctggagatt
     1201 aatgggcgga acaatctgat tcaacagaag actgccgcag ccatgttcgc ccagcagatg
     1261 cagcttatgc tccaaaacgc tcaaatgtca tcacttggtt cttttcctat gactccatca
     1321 attccagcta atcctcccat ggctttcaat ccttacatac cacatcctgg gatgggcctc
     1381 gttcctgcag aacttgtacc aaatacacct gttctgattc ctggaaaccc acctcttgca
     1441 atgccaggag ctgttggccc aaaactgatg cgttcagata aactggaggt ttgccgagaa
     1501 tttcagcgtg gaaattgtac ccgtggggag aatgattgcc gctatgctca ccctactgat
     1561 gcttccatga ttgaagcgag tgataatact gtgacaatct gcatggatta catcaaaggt
     1621 cgatgctcgc gggagaaatg caagtacttt catcctcctg cacacttgca agccagactc
     1681 aaggcagctc atcatcagat gaaccattca gctgcctctg ccatggccct gcagcctggt
     1741 acactgcaac tgataccaaa gagatcagca ctggaaaagc ccaatggtgc caccccggtc
     1801 tttaatccca ctgttttcca ctgccaacag gctctgacta acctgcagct cccacagccg
     1861 gcatttatcc ctgcagtgcc catgatgcac ggtgctacac ctaccactgt gtctgcagca
     1921 acaacacctg ccaccagcgt tccgttcgct gcaccaacta caggcaatca gctgaaattc
     1981 tgaacagcag agttgtcgac ggtaccgcgg gcccgtttaa acccgctgat cagcctcgac
     2041 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2101 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2161 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2221 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     2281 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     2341 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2401 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2461 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2521 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2581 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2641 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     2701 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     2761 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     2821 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2881 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     2941 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3001 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3061 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     3121 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     3181 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     3241 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     3301 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     3361 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     3421 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     3481 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     3541 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     3601 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     3661 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     3721 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     3781 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     3841 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     3901 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     3961 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     4021 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4081 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     4141 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4201 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     4261 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4321 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4381 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4441 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4501 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4561 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4621 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4681 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4741 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4801 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4861 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4921 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4981 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5041 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5101 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5161 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5221 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5281 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5341 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5401 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5461 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5521 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5581 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5641 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5701 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5761 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5821 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5881 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5941 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6001 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6061 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6121 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6181 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6241 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6301 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6361 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6421 cccgaaaagt gccacctgac gtc
//
