LOCUS       sequence-481944-        7335 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  inducible Mex3a fused to mouse Adar Deaminase domain (HyperTRIBE) 
            and V5 with t2a mCherry for expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7335)
  TITLE     Stemness factor Mex3a times translation and protein trafficking to 
            ensure robust differentiation of olfactory sensory neurons
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7335)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: tetO-Mex3a_HyperTRIBE-adar_deaminase-V5-t2a-mC...
FEATURES             Location/Qualifiers
     source          1..7335
                     /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             3272..3313
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3335..3388
                     /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             3395..4102
                     /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(3537..3555)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3805..3824
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          4124..4696
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(4828..4847)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4893..4948
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4947..4966)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5346..5363)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5517..6105)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5597..5616)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6276..7067,7068..7136))
                     /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     6899..6918
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7137..7241)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7309..7327
                     /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 gaggtggagg
     2101 tagtaccttc cacgaccaga tagctatgct gagccacagg tgcttcaatg ctctgaccaa
     2161 cagtttccag ccctccctgc tcggccgcaa gatcctggct gccattatta tgaaaagaga
     2221 tccagaggac atgggtgttg tcgtgagttt ggggacaggc aatcgctgtg tgaaagggga
     2281 ctctctcagc ctgaagggag agacggtcaa tgactgccat gccgaaatca tctcccggag
     2341 gggcttcatc aggtttctct acagtgaact gatgaagtac aaccaccaca ctgccaagaa
     2401 cagcatattt gagcttgcca ggggaggaga gaagctgcag ataaagaaga cggtttcttt
     2461 tcatctctac atcagcacgg cgccatgtgg agatggggcc ctctttgaca aatcctgcag
     2521 tgaccgtgcc gtggaaagca cagagtcccg ccattaccct gtctttgaaa atcccaagca
     2581 aggcaagctt cgcaccaagg tggagaatgg gcaaggcaca attcctgtgg agtccagtga
     2641 tattgtaccc acgtgggatg gcatccgcct tggggaaaga ctccgtacca tgtcctgtag
     2701 tgacaaaatc ctacgctgga atgtgctggg cctgcaaggg gcgttgttga cgcacttcct
     2761 acagcctgtg tacctgaaat ctgtaacctt aggttacctt ttcagccaag ggcatctgac
     2821 ccgtgctatt tgctgccgcg tgaccagaga tgggaaagca tttgaggatg gactacgcta
     2881 tccctttatt gtcaaccacc ccaaggtcgg ccgagtcagt gtttatgatt ccaaaagaca
     2941 gtccggaaag accaaggaaa caagcgtcaa ctggtgcatg gctgatggct atgacctaga
     3001 gatcctggat ggcaccagag gcactgtgga tggaccaggg aaagagttgt ctcgggtgtc
     3061 caagaagaat attttccttc agtttaagaa gctctgctcc ttccgagccc gcagagattt
     3121 actgcagctc tcttatggtg aagccaagaa agctgcccgt gactatgact tagccaagaa
     3181 ctacttcaag aaaagcctgc gagacatggg ctatgggaat tggatcagca aaccccagga
     3241 ggaaaagaac tttacggatc cccagccatg gggtaagcct atccctaacc ctctcctcgg
     3301 tctcgattct acgaagcttg tcgacggcag tggagagggc agaggaagtc tgctaacatg
     3361 cggtgacgtc gaggagaatc ctggcccatc tagagtgagc aagggcgagg aggataacat
     3421 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
     3481 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     3541 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
     3601 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
     3661 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
     3721 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
     3781 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc
     3841 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct
     3901 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
     3961 gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa
     4021 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
     4081 catggacgag ctgtacaagt aatctagagc tgagaacttc agggtgagtt tggggaccct
     4141 tgattgttct ttctttttcg ctattgtaaa attcatgtta tatggagggg gcaaagtttt
     4201 cagggtgttg tttagaatgg gaagatgtcc cttgtatcac catggaccct catgataatt
     4261 ttgtttcttt cactttctac tctgttgaca accattgtct cctcttattt tcttttcatt
     4321 ttctgtaact ttttcgttaa actttagctt gcatttgtaa cgaattttta aattcacttt
     4381 tgtttatttg tcagattgta agtactttct ctaatcactt ttttttcaag gcaatcaggg
     4441 tatattatat tgtacttcag cacagtttta gagaacaatt gttataatta aatgataagg
     4501 tagaatattt ctgcatataa attctggctg gcgtggaaat attcttattg gtagaaacaa
     4561 ctacaccctg gtcatcatcc tgcctttctc tttatggtta caatgatata cactgtttga
     4621 gatgaggata aaatactctg agtccaaacc gggcccctct gctaaccatg ttcatgcctt
     4681 cttctctttc ctacagctcc tgggcaacgt gctggttgtt gtgctgtctc atcattttgg
     4741 caaagaattc actcctcagg tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa
     4801 tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt atggggacat
     4861 catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat
     4921 agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt
     4981 aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata tgctggctgc
     5041 catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc ctgctgtcca
     5101 ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat tttgttttgt
     5161 gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag ccagattttt
     5221 cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga gatccctcga
     5281 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5341 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5401 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5461 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5521 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5581 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5641 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5701 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5761 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5821 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5881 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5941 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6001 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6061 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6121 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6181 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6241 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6301 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6361 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagaa
     6421 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6481 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6541 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6601 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6661 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6721 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6781 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6841 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat
     6901 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6961 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7021 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7081 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7141 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7201 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7261 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     7321 acgaggccct ttcgt
//
