LOCUS       sequence-441777-        6105 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tetO responsive expression of Mex3a in mammalian cells with t2a 
            cleaved mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6105)
  TITLE     Mex3a-dependent post-transcriptional silencing assures olfactory 
            receptor diversity and axon guidance specificity
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6105)
  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            "Unpublished"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: pTRE2_CMV_Mex3a_t2a_mCherry
FEATURES             Location/Qualifiers
     source          1..6105
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    join(20..38,39..61,62..80,81..103,104..122,123..145,
                     146..164,165..187,188..206,207..229,230..248,249..271,
                     272..290)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        323..361
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     358..382
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2105..2158
                     /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             2165..2872
                     /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(2307..2325)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2575..2594
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2894..3466
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(3598..3617)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3663..3718
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3717..3736)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4116..4133)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4287..4875)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4367..4386)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5046..5837,5838..5906))
                     /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     5669..5688
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5907..6011)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6079..6097
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cttcactcga gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca
       61 ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga
      121 gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt
      181 ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt
      241 gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag
      301 tcgagctcgg tacccgggtc gaggtaggcg tgtacggtgg gaggcctata taagcagagc
      361 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
      421 aagacaccgg gaccgatcca gcctccgcgg ccccgaattc gagctcggta cccggggatc
      481 ctctagtcag ctgacgcgtg ctagcgcggc catgcctagt ctagtggtat ctggaataat
      541 ggaaagaaat gggggctttg gagaactagg atgtttcggg ggaagcgcta aagacagggg
      601 gcttctggaa gacgagcgcg ccctccagct ggcactggac cagctctgcc ttctggggct
      661 gggcgagccg cctgcaccca ccgccggaga agatggtggg ggagggggcg gcggagcacc
      721 agcccagcct gcagcaccac ctcagccagc tccccctcct cctccagcag ccgctcccgc
      781 tgctccaagc gcagcccctg ccgctcaaac cccacaacct cccacagccc ctaagggcgc
      841 aaacgatgcc aagctgtgtg ccctgtataa agaggcagaa ttgaggctta aaggaagctc
      901 aaacaccacg gagtgcgttc ccgtgcccac ctccgagcac gtggccgaga tcgtgggcag
      961 gcaaggctgc aagattaagg ctctgagggc caaaaccaac acctacatca agacacctgt
     1021 gcgaggtgag gagccagtgt tcatggtgac tgggaggcgg gaagacgtgg ccacagcccg
     1081 gcgggagatc atctctgcag ctgagcactt ctccatgata agagcctcac gcaacaagtc
     1141 tggcgccgcc tttggcgtgg cccctgctct gccaggccaa gtgaccatcc gtgtacgagt
     1201 accctaccgc gtggtgggac tggtggtggg ccccaaggga gcgaccatca aacgcatcca
     1261 gcagcagacc aacacgtaca tcatcactcc cagccgggac cgcgaccccg tgtttgagat
     1321 aaccggcgca ccgggcaacg tggagcgtgc gcgggaggag atcgagacgc acatcgctgt
     1381 gcgcaccggc aagattcttg aatacaacag cgacggcgac ttcctggctg gcagccctga
     1441 cgctgccttg gacagccgct actcagacgc ctggcgggtg catgctccag gctgtaagcc
     1501 gctctccacc ttcaggcaga acagcctggg ctgcataggc gaatgcggcg tggactcggc
     1561 ctttgaggcc ccgcgcctaa gcgagcaggg cggggacttt ggctacagcg gatatctgtt
     1621 tccgggctac ggtgtgggca agcaggacgt gtactatggc gtggcagaga caagcccccc
     1681 gctgtgggcg ggtcaggaga atgccacgcc cacctcggtg ctcttctcct ctgcctcctc
     1741 ctcctcctct tcagccaagg cacgcgcggg ccccccaggc gcgcatcgct ctcctgccac
     1801 ctccgcgggg ccagagcagc tgaccgggct ccctcggagg ccacccggag agccactgca
     1861 gggtttctct aaactgggga caggcgggct gcggagcccc ggcagcgggc gggactgcat
     1921 ggtgtgcttt gagagtgaag tgacagcagc cctggtgccc tgtggacaca accttttctg
     1981 catggagtgt gcagtacgca tctgcgagag gacggaccca gagtgtcctg tctgccacat
     2041 caccgccaca caagccatcc gaatattttc cggccgcatc gataagcttg tcgacggcag
     2101 tggagagggc agaggaagtc tgctaacatg cggtgacgtc gaggagaatc ctggcccatc
     2161 tagagtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
     2221 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     2281 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     2341 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     2401 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     2461 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     2521 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     2581 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     2641 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     2701 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     2761 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     2821 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt aatctagagc
     2881 tgagaacttc agggtgagtt tggggaccct tgattgttct ttctttttcg ctattgtaaa
     2941 attcatgtta tatggagggg gcaaagtttt cagggtgttg tttagaatgg gaagatgtcc
     3001 cttgtatcac catggaccct catgataatt ttgtttcttt cactttctac tctgttgaca
     3061 accattgtct cctcttattt tcttttcatt ttctgtaact ttttcgttaa actttagctt
     3121 gcatttgtaa cgaattttta aattcacttt tgtttatttg tcagattgta agtactttct
     3181 ctaatcactt ttttttcaag gcaatcaggg tatattatat tgtacttcag cacagtttta
     3241 gagaacaatt gttataatta aatgataagg tagaatattt ctgcatataa attctggctg
     3301 gcgtggaaat attcttattg gtagaaacaa ctacaccctg gtcatcatcc tgcctttctc
     3361 tttatggtta caatgatata cactgtttga gatgaggata aaatactctg agtccaaacc
     3421 gggcccctct gctaaccatg ttcatgcctt cttctctttc ctacagctcc tgggcaacgt
     3481 gctggttgtt gtgctgtctc atcattttgg caaagaattc actcctcagg tgcaggctgc
     3541 ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct
     3601 ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg
     3661 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     3721 cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag
     3781 tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca
     3841 tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg
     3901 aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt
     3961 ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc
     4021 tgtccctctt ctcttatgga gatccctcga ctgcattaat gaatcggcca acgcgcgggg
     4081 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4141 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4201 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4261 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4321 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4381 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4441 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4501 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4561 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4621 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4681 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4741 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4801 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4861 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4921 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4981 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5041 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5101 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5161 ggccccagtg ctgcaatgat accgcgagaa ccacgctcac cggctccaga tttatcagca
     5221 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5281 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5341 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5401 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5461 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5521 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5581 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5641 agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa
     5701 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5761 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5821 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5881 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5941 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6001 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     6061 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgt
//
