LOCUS       sequence-441774-        5784 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mutant RING Mex3a in mammalian cells with t2a cleaved 
            mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5784)
  TITLE     Mex3a-dependent post-transcriptional silencing assures olfactory 
            receptor diversity and axon guidance specificity
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5784)
  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: 
            "Unpublished"
COMMENT     Sequence Label: CMV_Mex3a_mutRING_t2a_mCherry
FEATURES             Location/Qualifiers
     source          1..5784
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        17..55
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     52..76
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1799..1852
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1859..2566
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2001..2019)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2269..2288
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2588..3160
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(3292..3311)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3357..3412
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3411..3430)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3810..3827)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3981..4569)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4061..4080)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4740..5531,5532..5600))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5363..5382
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5601..5705)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(5773..5784,1..7)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 acgaggccct ttcgtcggta ggcgtgtacg gtgggaggcc tatataagca gagctcgttt
       61 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca
      121 ccgggaccga tccagcctcc gcggccccga attcgagctc ggtacccggg gatcctctag
      181 tcagctgacg cgtgctagcg cggccatgcc tagtctagtg gtatctggaa taatggaaag
      241 aaatgggggc tttggagaac taggatgttt cgggggaagc gctaaagaca gggggcttct
      301 ggaagacgag cgcgccctcc agctggcact ggaccagctc tgccttctgg ggctgggcga
      361 gccgcctgca cccaccgccg gagaagatgg tgggggaggg ggcggcggag caccagccca
      421 gcctgcagca ccacctcagc cagctccccc tcctcctcca gcagccgctc ccgctgctcc
      481 aagcgcagcc cctgccgctc aaaccccaca acctcccaca gcccctaagg gcgcaaacga
      541 tgccaagctg tgtgccctgt ataaagaggc agaattgagg cttaaaggaa gctcaaacac
      601 cacggagtgc gttcccgtgc ccacctccga gcacgtggcc gagatcgtgg gcaggcaagg
      661 ctgcaagatt aaggctctga gggccaaaac caacacctac atcaagacac ctgtgcgagg
      721 tgaggagcca gtgttcatgg tgactgggag gcgggaagac gtggccacag cccggcggga
      781 gatcatctct gcagctgagc acttctccat gataagagcc tcacgcaaca agtctggcgc
      841 cgcctttggc gtggcccctg ctctgccagg ccaagtgacc atccgtgtac gagtacccta
      901 ccgcgtggtg ggactggtgg tgggccccaa gggagcgacc atcaaacgca tccagcagca
      961 gaccaacacg tacatcatca ctcccagccg ggaccgcgac cccgtgtttg agataaccgg
     1021 cgcaccgggc aacgtggagc gtgcgcggga ggagatcgag acgcacatcg ctgtgcgcac
     1081 cggcaagatt cttgaataca acagcgacgg cgacttcctg gctggcagcc ctgacgctgc
     1141 cttggacagc cgctactcag acgcctggcg ggtgcatgct ccaggctgta agccgctctc
     1201 caccttcagg cagaacagcc tgggctgcat aggcgaatgc ggcgtggact cggcctttga
     1261 ggccccgcgc ctaagcgagc agggcgggga ctttggctac agcggatatc tgtttccggg
     1321 ctacggtgtg ggcaagcagg acgtgtacta tggcgtggca gagacaagcc ccccgctgtg
     1381 ggcgggtcag gagaatgcca cgcccacctc ggtgctcttc tcctctgcct cctcctcctc
     1441 ctcttcagcc aaggcacgcg cgggcccccc aggcgcgcat cgctctcctg ccacctctgc
     1501 ggggccagag cagctgaccg ggctccctcg gaggccaccc ggagagccac tgcagggttt
     1561 ctctaaactg gggacaggcg ggctgcggag ccccggcagc gggcgggact gcatggtgtg
     1621 ctttgagagt gaagtgacag cagccctggt gcccgctgga gccaaccttt tcgctatgga
     1681 ggctgcagta cgcatctgcg agaggacgga cccagagtgt cctgtctgcc acatcaccgc
     1741 cacacaagcc atccgaatat tttccggccg catcgataag cttgtcgacg gcagtggaga
     1801 gggcagagga agtctgctaa catgcggtga cgtcgaggag aatcctggcc catctagagt
     1861 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
     1921 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     1981 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
     2041 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
     2101 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     2161 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
     2221 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
     2281 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
     2341 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt
     2401 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat
     2461 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
     2521 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtaatcta gagctgagaa
     2581 cttcagggtg agtttgggga cccttgattg ttctttcttt ttcgctattg taaaattcat
     2641 gttatatgga gggggcaaag ttttcagggt gttgtttaga atgggaagat gtcccttgta
     2701 tcaccatgga ccctcatgat aattttgttt ctttcacttt ctactctgtt gacaaccatt
     2761 gtctcctctt attttctttt cattttctgt aactttttcg ttaaacttta gcttgcattt
     2821 gtaacgaatt tttaaattca cttttgttta tttgtcagat tgtaagtact ttctctaatc
     2881 actttttttt caaggcaatc agggtatatt atattgtact tcagcacagt tttagagaac
     2941 aattgttata attaaatgat aaggtagaat atttctgcat ataaattctg gctggcgtgg
     3001 aaatattctt attggtagaa acaactacac cctggtcatc atcctgcctt tctctttatg
     3061 gttacaatga tatacactgt ttgagatgag gataaaatac tctgagtcca aaccgggccc
     3121 ctctgctaac catgttcatg ccttcttctc tttcctacag ctcctgggca acgtgctggt
     3181 tgttgtgctg tctcatcatt ttggcaaaga attcactcct caggtgcagg ctgcctatca
     3241 gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc
     3301 ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata
     3361 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
     3421 gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc
     3481 aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta
     3541 tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta
     3601 gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt
     3661 acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc
     3721 tcttctctta tggagatccc tcgactgcat taatgaatcg gccaacgcgc ggggagaggc
     3781 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3841 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3901 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3961 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4021 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4081 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4141 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4201 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4261 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4321 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4381 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4441 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4501 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4561 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4621 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4681 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4741 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4801 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4861 agtgctgcaa tgataccgcg agaaccacgc tcaccggctc cagatttatc agcaataaac
     4921 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4981 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5041 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5101 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5161 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5221 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5281 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5341 tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5401 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5461 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5521 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5581 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5641 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5701 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     5761 ttaacctata aaaataggcg tatc
//
